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Satellite DNAs (satDNAs) are a major component of eukaryotic genomes, playing key roles in chromosome organization, heterochromatin formation, and genome evolution. Although satellitomes have been increasingly characterized in insects, most studies focus on monocentric species, leaving holocentric systems comparatively understudied. Here, we investigate the satellitomes of two pentatomid bugs, Edessa meditabunda and Edessa loxdalii, which exhibit unusually large heterochromatin blocks for holocentric chromosomes. Using an integrative approach combining genome analysis and molecular cytogenetics, we identified 31 satDNA families shared between species. E. loxdalii shows higher satDNA abundance and lower sequence divergence, consistent with recent amplification and homogenization, whereas E. meditabunda exhibits lower abundance and higher divergence, suggesting more advanced sequence degeneration. SatDNA landscape analyses indicate distinct evolutionary trajectories, with evidence of multiple amplification waves and differential repeat turnover. Despite these differences, both species share a conserved set of satDNAs, including highly abundant families that dominate their satellitomes, supporting the library hypothesis. Compared to other Pentatomidae species with low heterochromatin content, there is no overall increase in satDNA abundance in Edessa, but rather amplification of specific families. Chromosomal mapping revealed that these amplified satDNAs are major components of terminal heterochromatin blocks, while less abundant families show chromosome-specific distributions, including accumulation on the Y chromosome, with signs of evolutionary differentiation between species. Overall, this study provides new insights into satellitome organization and evolution in Pentatomidae, highlighting the role of satDNAs in shaping chromosome architecture in holocentric systems.

<h4>Background</h4>Helicobacter pylori (H. pylori) infection is the principal etiological factor for gastric cancer (GC), which ranks as the fourth leading cause of cancer-related mortality globally. Although the Correa cascade describes the stepwise progression from chronic gastritis to intestinal-type GC, the molecular events underlying this "inflammation-to-cancer" transition remain incompletely characterized.<h4>Objective</h4>This study aimed to delineate stage-specific proteomic alterations across H. pylori-associated gastric mucosal lesions and to identify candidate progression-associated biomarkers in GC.<h4>Methods</h4>Label-free quantitative proteomic analysis (LC-MS/MS) was performed on gastric tissue specimens from 22 patients stratified into four histopathological stages: H. pylori-negative chronic non-atrophic gastritis (HpN-CG), H. pylori-positive chronic non-atrophic gastritis (HpC-CG), H. pylori-positive chronic atrophic gastritis (HpC-CAG), and intestinal-type GC. Functional enrichment (GO, KEGG), temporal clustering (Mfuzz), and protein-protein interaction (PPI) network analyses were conducted. Key hub genes were externally assessed using GEPIA RNA-seq data from TCGA and GTEx.<h4>Results</h4>A total of 6,019 proteins were quantified. Differential expression analysis identified 771 DEPs between HpC-CG and HpN-CG, 101 DEPs between HpC-CAG and HpC-CG, and 535 DEPs between GC and HpC-CAG. Immune response pathways were up-regulated upon H. pylori infection, whereas oxidative phosphorylation was progressively suppressed throughout disease progression. Mfuzz clustering revealed that Cluster 4 ribosome-biogenesis proteins, including NIP7 and PDCD11 identified in the proteomic/PPI analysis, exhibited continuous up-regulation from precancerous stages to GC. External RNA-expression validation supported significant up-regulation of seven Cluster 4 hub genes, whereas Cluster 6 proteins did not show concordant transcript-level down-regulation.<h4>Conclusion</h4>This study provides a comprehensive proteomic landscape of H. pylori-driven gastric carcinogenesis. Cluster 4 ribosome-biogenesis proteins represent candidate progression-associated protein markers requiring further protein-level validation, while the progressive decline in oxidative phosphorylation underscores mitochondrial dysfunction as a hallmark of disease progression.

<h4>Background</h4>Understanding how skeletal muscle responds to weight loss is crucial for developing targeted strategies to manage obesity and promote sustained improvement in metabolic health. Here, we investigated the molecular mechanisms underlying skeletal muscle reprogramming of gene expression and metabolic activity following Roux-en-Y gastric bypass (RYGB).<h4>Methods</h4>Forty-one women were studied before and one year after RYGB surgery. We leveraged multi-omics (DNA methylomics and transcriptomics) and machine learning approaches to complement muscle metabolic analyses and clinical data to identify mechanisms underlying RYGB-induced muscle metabolic reprogramming.<h4>Findings</h4>RYGB markedly decreased body weight and fat mass and improved metabolic health. Integrative analysis of vastus lateralis muscle identified 8233 genes with differentially methylated regions and 2173 differentially expressed genes post-RYGB surgery, of which 1197 genes were both differentially methylated and differentially expressed. Promoter hypomethylation was associated with the enhanced expression of transcription factors involved in skeletal muscle development and ribosomal subunits. In contrast, expression of genes encoding mitochondrial proteins decreased despite increases in mitochondrial content and enhanced mitochondrial function in skeletal muscle post-RYGB. Pre-operative muscle OXPHOS capacity, and expression of skeletal muscle hypertrophy and differentiation genes MYOC and EHMT2 were associated with weight loss success.<h4>Interpretation</h4>RYGB improves systemic metabolic health and induces sustained skeletal muscle bioenergetic reprogramming characterised by enhanced expression of genes involved in myogenesis and protein translation, but decreased expression of genes involved in mitochondrial metabolism, which may reflect improved mitochondrial quality and function. These findings advance our understanding of skeletal muscle metabolic responses to weight loss and of individual variability in metabolic phenotypes.<h4>Funding</h4>Canadian Institutes of Health Research (CIHR PJT183651-M-EH, 201709FDN-CEBA-116200-GRS), Diabetes Canada Investigator Award grant OG-3-22-5645-GS (GRS), J. Bruce Duncan Endowed Chair in Metabolic Diseases (GRS), Tier 1 Canada Research Chair in Mitochondrial Bioenergetics and Metabolic Health (M-EH), Tier 1 Canada Research Chair in Metabolic Diseases (GRS).

Ovarian cancer is often diagnosed at an advanced stage because its early symptoms are nonspecific, and the prognosis remains poor due to frequent recurrence and the development of drug resistance even after standard treatment. This review includes 19 <i>in vitro</i> studies and seeks to mechanistically summarize how reactive oxygen species (ROS) reinforce the tumorigenic program from the early stages of ovarian carcinogenesis to tumor progression, metastasis, and treatment resistance. Collectively, the selected studies suggest that exposure to environmental toxicants can increase ROS levels in ovarian epithelial cells, accompanied by inflammatory responses and elevated DNA damage markers, thereby creating a carcinogenesis-priming environment. During the progression stage, hypoxia, hormones, growth factors, and lipid signaling repeatedly activate survival and growth pathways, such as those involving HIF-1α/VEGF, JAK/STAT3, and AKT/mTOR, through ROS-mediated mechanisms to promote tumor cell proliferation, anti-apoptotic activity, and angiogenesis. In addition, ROS are associated with alterations in epithelial-to-mesenchymal transition-related markers, remodeling of the extracellular matrix, and regulation of matrix metalloproteinases, which enhance the metastatic and invasive potential of tumor cells. In the therapeutic context, ROS have been suggested to contribute to platinum-based chemotherapy resistance through mechanisms including changes in mitochondrial dynamics, activation of the DNA damage response, and reprogramming of ROS-dependent phosphorylation networks. Understanding the diverse molecular mechanisms and clinical manifestations associated with ROS expression in high-grade serous ovarian cancer (HGSOC) will contribute to a more precise understanding of its pathophysiology. Furthermore, studies on redox-targeted therapeutic strategies, particularly the application of mitochondria-targeted antioxidants, may provide valuable translational insights for the treatment of ovarian cancer and other ROS-related diseases.

Expansion of CAG repeats in HTT exon 1 is the acknowledged driver of Huntington's disease. Alternative processing of HTT pre-mRNA generates the truncated HTT1a transcript, translated into a toxic peptide. While its dependence on CAG length is well documented, the role of adjacent sequences - particularly the Proline-Rich Domain (PRD) - remains unexplored. Using our HuntEx1-engineered mouse embryonic stem cell platform, we show that human PRD promotes HTT1a production, whereas its replacement with mouse PRD in an otherwise human exon 1 markedly reduces HTT1a levels. Mechanistically, we find that the PRD shapes mRNA structure, and motif analysis identifies Serine-Arginine Splicing Factor 7 (SRSF7) binding sites in mouse but not in human PRD. Their targeted mutation confirms SRSF7's regulatory role in suppressing HTT1a production. Our findings establish the PRD as a key cis-regulator of HTT1a biogenesis, demonstrating that HTT toxicity also depends on sequence context, and highlighting splicing-based, PRD-focused therapeutic avenues.

<title>Abstract</title> <p> <bold>Background</bold> Genetic studies are conducted in athletic populations to understand the influence of genotype on phenotype, with the prospects of identifying talent, designing individualized training programmes, and predicting injuries in sports. Despite extensive research on improving water sports performance, the effects of various genetic polymorphisms on competitive performance remain unclear. <bold>Objective</bold> This systematic review aims to identify key gene variants associated with on-water sports and assess their influence on performance. <bold>Methods</bold> To fulfill this aim, a systematic search was conducted in Web of Science, Scopus, and PubMed until 6th October 2024, along with a secondary search of the included full-text articles. Studies examining the associations between genetic variants and on-water sports performance (rowing, canoeing, kayaking, sailing, surfing, and rafting) and meeting the Hardy‒Weinberg equilibrium criteria were included, and the risk of bias was determined. <bold>Results</bold> A total of 28 articles (6 high risk-of-bias) were included and reviewed (22 rowing, 4 kayaking, 5 canoeing, 1 each on surfing and rafting and none on sailing) that examined 17 different variants (ACE, ACTN3, AMPD1, CKMM, GABPB1, GDF8, HFE, HIF1A, MCT1, NOS3, PPARA, PPARGC1A, TFAM, UCP2, UCP3, VEGFR, and VEGFR2) from 8 different populations (Russians, Polish, Spanish, Brazilians, Hungarian, Australian, Turkish, and Chinese) associated with performance. The available evidence was inadequate for performing a meta-analysis and determining whether any of the genetic variants identified in this review were associated with sports performance. <bold>Conclusion</bold> Although several structural and functional genes and their polymorphisms have been identified that are associated with on-water sports performance, further collaborative genome-wide association studies and gene expression and gene‒gene interaction studies are needed in larger populations of different ethnicities to investigate additional possible genetic variants and the role of these gene polymorphisms in enabling success in sports. </p>

<title>Abstract</title> <p>Lithium is the gold standard mood stabiliser used to treat cycling mania and depression in bipolar disorder. Despite seven decades of clinical use, the mechanisms of its mood stabilisation are incompletely understood, fundamentally limiting development of improved alternatives. Two established lithium targets, glycogen synthase kinase 3β (GSK3β) and inositol monophosphatase, both modulate phosphorylation, suggesting lithium may exert broad effects on neuronal phosphorylation networks. We performed a discovery-phase in vitro screen of 140 kinases at 10mM LiCl and demonstrated that lithium inhibits 17 kinases beyond GSK3β. We therefore used untargeted quantitative phosphoproteomics to create a comprehensive map of lithium's phosphorylation signature in mouse synaptoneurosomes collected at dawn and dusk, matching peaks in phosphorylation driven by the sleep/wake cycle. Genes encoding lithium-sensitive phosphoproteins were significantly enriched in bipolar disorder genome-wide association studies, providing independent genomic evidence that these phosphorylation networks are relevant to bipolar pathophysiology. We further refined existing models of lithium’s action by showing that GSK3β inhibition is temporally restricted to dawn, indicating cross talk with sleep/wake cycles of phosphorylation. Overall, our data demonstrate that lithium’s pleiotropic effects may result from coordinated multi-kinase network reorganisation rather than single-target inhibition — a principle with direct implications for rational polypharmacology in mood stabiliser development.</p>

Importin 7 (IPO7) is a nuclear transport receptor of the β-karyopherin family that mediates the translocation of a broad spectrum of macromolecules, commonly referred to as cargoes. Discovered nearly three decades ago, IPO7 was initially identified as an import receptor for constitutive cellular cargoes, including histone H1 and ribosomal proteins, and was shown to function synergistically and partially redundantly with canonical receptors such as importin β1 and karyopherin β2. Over the past 15 years, however, accumulating evidence has established IPO7 as an important mediator of signal-dependent nuclear trafficking in response to extracellular stimuli, including cytokines, growth factors, and cellular stress. Thus, IPO7 has emerged as a versatile nuclear transport receptor that couples extracellular signaling to dynamic changes in nuclear composition and gene expression. Mechanistically, many IPO7 cargoes lack classical nuclear localization signals and instead contain noncanonical targeting motifs that directly engage IPO7. In several cases, phosphorylation-dependent conformational changes expose these motifs, promoting IPO7 binding and translocation through the nuclear pore complex. The expanding repertoire of IPO7 cargoes, including ERK, SMAD3, EGR1, GLI1, the glucocorticoid receptor, HIF-1α, YAP1, and RUNX2, highlights its prominent role at the interface of signaling and transcriptional control. Consistent with these functions, dysregulation of IPO7-mediated transport has been implicated in cancer, hypoxia, and other pathological states. Beyond cellular signaling, IPO7 also contributes to the nuclear trafficking of viral genomes and proteins. Here, we review the molecular mechanisms of IPO7-dependent nuclear import, emerging principles of cargo recognition, and pathways that regulate IPO7 activity during cellular signaling.

<h4>Background</h4>Idiopathic pulmonary fibrosis (IPF) is a progressive interstitial lung disease in which both environmental exposures and genetic predisposition contribute to disease susceptibility. Studying the burden of rare variants in a genetically homogeneous founder population may help identify disease-associated alleles that are difficult to detect in more heterogeneous populations.<h4>Methods</h4>Whole exome sequencing was performed on 123 patients with IPF and 1110 unrelated controls from Sardinia (Italy). Variant prioritisations were conducted according to the American College of Medical Genetics (ACMG) guidelines. In parallel, gene burden of rare predicted loss-of-function variants was assessed using the Cohort Allelic Sums Test (CAST) algorithm to identify genes significantly enriched in IPF cases compared with controls.<h4>Results</h4>Pathogenic or likely pathogenic variants in known telomere-related genes were identified in 11.4% of patients. These variants were associated with younger age at diagnosis and a higher prevalence among never-smokers. CAST analysis identified a significant enrichment of loss-of-function variants in 82 genes, including <i>MUC5B</i> (OR=443.1, false discovery rate=3.7E-05), functionally related to cilium organisation and motility, with a significant overrepresentation of dynein-related genes.<h4>Conclusions</h4>This study supports the contribution of telomere-related variants to IPF susceptibility and suggests a possible role for rare variants affecting mucociliary pathways. Together, these findings broaden the current understanding of IPF biology in this cohort. The distinctive genetic background of the Sardinian population may have facilitated the identification of rare or population-specific variants, underscoring the potential value of founder populations in complex disease genetics.

Pancreatic ductal adenocarcinoma (PDAC) is among the most lethal cancers, with over 83% of patients presenting metastases to vital organs such as the liver, lungs, and peritoneum. Although glycosylation has been established as a critical factor in PDAC development, little is known about the molecular underpinnings of organ-specific metastasis. In this study, we investigated the role of glycosyltransferases (GTs) in mediating PDAC metastatic organotropism. Through an unbiased transcriptomic screen, we identified distinct GT expression patterns in liver- and lung-tropic PDAC cells. Notably, GCNT3 and B3GNT3 were selectively upregulated in liver and lung-tropic cells, respectively. These mutually exclusive expression patterns were validated in human metastatic PDAC specimens, where GCNT3 was enriched in liver metastatic lesions and B3GNT3 in lung metastases. Proteomic profiling revealed that differential expression of these GTs is associated with epithelial-mesenchymal transition and glycoproteome remodeling in metastatic cells. Functional studies using in vitro assays, ex vivo organotypic cultures, and in vivo metastasis models demonstrated that loss of GCNT3 or B3GNT3 impairs the proliferation and survival of PDAC cells in their respective metastatic organ environments. Together, these findings uncover a glycosylation-based mechanism that facilitates organ-specific survival of metastatic PDAC cells and highlight glycosyltransferases as potential therapeutic targets for disrupting metastatic adaptation.

Huntington's disease [HD] is a progressive, autosomal dominant neurodegenerative disorder caused by a pathogenic CAG repeat expansion in the HTT gene, resulting in mutant huntingtin [mHTT] protein accumulation, neuronal dysfunction, and selective neurodegeneration. Current pharmacological management remains largely symptomatic, with no approved therapies capable of modifying disease progression. In recent years, however, significant advances in molecular neuroscience and translational medicine have accelerated the development of disease-modifying strategies targeting the underlying pathogenic mechanisms of HD. This review synthesizes emerging pharmacological therapies with a particular focus on insights derived from recent and ongoing clinical trials. Key therapeutic approaches discussed include gene-silencing technologies such as antisense oligonucleotides, RNA interference, and CRISPRCas9- based strategies, as well as small-molecule modulators targeting mutant huntingtin aggregation, proteostasis, autophagy, mitochondrial dysfunction, and neuroinflammation. In addition, advances in symptomatic treatments addressing motor, cognitive, and psychiatric manifestations are reviewed. The article critically examines translational challenges encountered in clinical development, including blood-brain barrier penetration, allele selectivity, dosing paradigms, patient heterogeneity, biomarker integration, and ethical considerations associated with irreversible genetic interventions. Lessons learned from both successful and failed trials highlight the importance of precision medicine approaches, biomarker-guided trial designs, and combination therapies targeting multiple pathogenic pathways. Collectively, this review provides an updated and clinically relevant overview of the evolving HD therapeutic landscape and outlines key considerations for translating molecular advances into effective and safe pharmacological interventions.

Proton therapy spares normal tissues better than photon therapy, potentially reducing toxicity while maintaining tumor control. However, challenges remain due to variations in proton dose distribution, particularly at the distal edge of the spread-out Bragg peak (SOBP); for organs at risk, such variation is crucial. We evaluated the biological effects by comparing two positions of the proton profile, the entrance plateau (EP) and SOBP, in a murine model and investigated distinct metabolomic and proteomic signatures. Mice exposed to the EP beam segment (LET_d = 0.8 keV/µm) exhibited less weight reduction than their SOBP-irradiated counterparts (LET_d = 2.6 keV/µm). Two hours post-irradiation, the SOBP caused more severe DNA damage in the hippocampus and thalamus. Hematoxylin and eosin staining revealed eosinophil aggregation in both groups, with more surviving neurons in the EP group. Metabolomic profiles differed more between the EP and SOBP groups at 2 h than at 3 days post-irradiation. Relative to EP, SOBP irradiation at 2 h increased fructose-1,6-bisphosphate (FBP), dihydroxyacetone phosphate (DHAP), and inosine but decreased prostaglandin F2α; subsequently, proteomic analysis at day 3 showed that calcium signaling, NF-κB, and endocytosis pathways were enriched in the SOBP group. Combined multi-omics analysis further demonstrated that SOBP irradiation significantly activated the pentose phosphate pathway, purine metabolism, and phospholipase D signaling, while concurrently suppressing arachidonic acid metabolism. Our findings underscore the need for early detection of proton-induced brain toxicity and demonstrate that the higher-LET SOBP segment causes more severe damage than the EP. Targeting these dysregulated multi-omics pathways may offer a promising strategy for mitigating radiation-induced brain injury during proton therapy.

We report the first documented case of extreme, intact parathyroid hormone (PTH)-independent hypercalcemia mediated by a rare non-osteoclastic bone remodeling mechanism in a 65-year-old Puerto Rican woman, detected within 24 h of the second dose of the Moderna mRNA-1273 COVID-19 vaccine and leading to the diagnosis of high-risk biclonal IgA plasma cell myeloma (R-ISS Stage III). Peak serum calcium reached 17.4 mg/dL with concurrent hyperphosphatemia, normal lactate dehydrogenase (LDH), and without marrow osteoclasts or osteolytic lesions, findings inconsistent with classical cancer-associated hypercalcemia. Hypercalcemia resolved within eight days with supportive treatment alone, remaining durably normal nine months later. The temporal proximity to vaccination, together with the biochemical-histopathological profile, generates the hypothesis, without implying causation, that a vaccine-induced innate immune response triggered a rare inflammatory osteocytic perilacunar/canalicular remodeling. The biclonal gammopathy, Clone-1 (normal karyotype, CD33⁺, OCT-2⁺, MYC^-, CD19⁺, PAX5^-) and Clone-2 (complex high-risk karyotype, OCT-2^-, MYC⁺low, CD19^-), suggests epigenetic rather than genomic drivers in the predominant clone. This case proposes a novel mechanistic hypothesis for vaccine-associated hypercalcemia and generates testable questions that warrant prospective investigation.

Mechanisms governing initiation steps of the assembly of endogenous multi-protein complexes (EMC) remain incompletely understood. Here, multiple lines of observations are reported describing the function-aligned initiation sequence of hybrid assembly pathways (HAP) of EMC. The first step of HAP-guided chain reactions of protein-protein interactions (PPI) of EMC assemblies constitutes the creation of cell type-specific pools of hetero and homo dimers. The molecular anatomy of HAP was elucidated by defining qualitative and quantitative characteristics of protein binding to a compendium of 200,393 distinct genomic regulatory elements (GRE), including 49,667 sequences representing control sets of genomic loci as well as 150,726 GRE of different evolutionary origins. The consensus sequence of HAP actions consists of: a) Initiation on genomic DNA of the formation of metastable hetero- and homodimers of EMCs' protein constituents; b) Release of dimers from DNA templates for delivery to the EMC assembly compartments; c) Assembly of defined EMC by sequential on demand addition of proteins to preformed dimers serving as attractors of EMC-specific ensembles of monomers. Chromosome-naïve DNA scaffolds facilitating creation of intracellular dimer pools engage networks of ~700 transcription factors (TFs), 534 of which manifest region-specific patterns of significantly enriched expression in 1358 brain regions. HAP initiators appear to operate within nucleosome-depleted islands of transposable elements (TE) - derived sequences within heterochromatin. PPI assembly lines of EMCs operate in 2 concurrent modes: TF-TF PPI cascade and PPI HUB protein cascade. Regardless of the number of DNA-bound initiator TFs (ranging from one to 716 TFs), both modes of operations reached the equilibrium at the PPI constituents saturation levels of ~245 proteins for TF-TF PPI modes and of ~351 proteins for PPI HUB protein modes. Distinct panels of DNA-bound initiator TFs and proteins of PPI cascade ensembles are enriched in either defined sets of neuroanatomical structures (TF-TF mode) or among structural-functional constituents of synapses (HUB proteins mode). Thus, these bifurcated cascades appear biologically congruent: TF-TF constituents map to transcriptional signatures of hundreds of brain regions, whereas HUB constituents map to synaptogenesis and synaptic structures, suggesting the unified logic of genomic functions coordinating region identity and connectivity. Evidence-supported examples of default operations of PPI-guided assemblies of hetero- and homodimers of Yamanaka factors, neurogenesis constituents, and protein components of postsynaptic density of excitatory and inhibitory synaptogenesis are reported with detailed analytical focus on human Claustrum. The foundational set of observations reported in this contribution should facilitate experimental and theoretical explorations of TE-seeded genomic codes for initiators of PPI chain reactions of protein dimerization creating pools of attractors to guide and accelerate the EMC assemblies.

Members of the immunoglobulin superfamily (IgSF) are cell-surface proteins essential for a wide range of biological processes, including molecular transport, morphoregulation, and cell differentiation. IGSF10, a member of this family, has been identified as a disease-associated gene in delayed puberty and cleidocranial dysplasia. In this study, a Chinese patient with delayed puberty, microcephaly, scoliosis, and epilepsy was investigated. The genetic defect and immune alterations were assessed using whole-exome sequencing, Sanger sequencing, and single-cell RNA sequencing. After variant filtering, a novel IGSF10 variant, NM_178,822.5:c.4518del (p.His1506Gln fs Ter13), was identified in the proband. Sanger sequencing indicated that this variant was <i>de novo</i>, as it was present only in the proband and absent from her parents. Quantitative real-time PCR showed that the newly identified variant was associated with reduced IGSF10 mRNA abundance, consistent with nonsense-mediated decay. Single-cell RNA sequencing of peripheral blood mononuclear cells revealed that this variant was associated with marked alterations in effector T-cell and monocyte populations. Further analysis suggested altered differentiation patterns of T cells and monocytes, supporting the hypothesis that immune perturbations may contribute to the patient's reproductive, neurological, and skeletal phenotypes, although this possibility requires functional validation. A novel IGSF10 variant was identified in a Chinese patient with delayed puberty, microcephaly, scoliosis, and epilepsy. This case expands the mutational spectrum of IGSF10 deficiency and suggests that its phenotypic spectrum may include scoliosis and epilepsy, pending further confirmation. These findings also provide preliminary insight into the potential interplay among IGSF10, immune function, hormone secretion, and brain and skeletal development.

Alzheimer's disease (AD) is a neurodegenerative disorder characterized primarily by progressive cognitive impairment, whose pathogenesis involves multiple pathological processes including protein deposition, metal homeostasis dysregulation, oxidative stress, mitochondrial dysfunction, and neuroinflammation. In recent years, metabolism-related cell death modalities such as ferroptosis, cuproptosis, and disulfidptosis have gradually been recognized as potentially involved in neuronal damage in Alzheimer's disease. This review summarizes the fundamental mechanisms of ferroptosis, cuproptosis, and disulfidptosis, along with their research evidence in AD. Ferroptosis is primarily driven by iron imbalance, lipid peroxidation buildup, and impaired GPX4 defense. This process exhibits a bidirectional amplification loop with Aβ and tau pathologies. Cuproptosis contributes to neuronal damage through abnormal copper accumulation, FDX1-related mitochondrial protein lipoylation dysfunction, loss of iron-sulfur cluster proteins, and proteotoxic stress. Disulfidptosis links glucose metabolism disorders, insufficient reducing power, and actin cytoskeleton vulnerability, providing novel insights into metabolic stress and structural damage in AD. Furthermore, the three modes of cell death can undergo cross-regulation through the SLC7A11-NADPH-GSH/GPX4 axis, the FDX1-DLAT/DLST-iron-sulfur cluster axis, as well as upstream factors such as p53, NRF2, and AMPK. Metabolic cell death may constitute a critical pathological network in AD. Targeting these death pathways and their shared hubs is expected to provide new directions for disease stratification, biomarker development, and disease-modifying therapies.

<h4>Objective</h4>The objective of this study was to gain insight into the molecular mechanisms leading to early degeneration of decellularized homografts.<h4>Methods</h4>Formalin-fixed and paraffin-embedded tissues from fresh explanted decellularized aortic (<i>n</i> = 7) and pulmonary (<i>n</i> = 8) valves were used. RNA was isolated and analyzed using panel-based transcriptomics, focusing on fibrosis- and inflammation-related genes. Differentially expressed genes were used as input parameters for biological pathway analysis using the Gene Ontology Biological Process and Hallmark databanks. Formalin-fixed and paraffin-embedded tissues from freshly explanted healthy donor aortic (<i>n</i> = 7) and pulmonary valves (<i>n</i> = 8) were used as controls.<h4>Results</h4>Our analysis revealed 56 differentially expressed genes in decellularized aortic valves compared to donor aortic valves, of which 21 and 35 were up- and downregulated, respectively. Decellularized pulmonary valves showed 115 differentially expressed genes compared to donor pulmonary valves, 66 of which were up- and 49 downregulated. In both decellularized aortic and pulmonary valve explants, we found increased expression of fibrosis-, inflammation-, and endothelium-related genes and a decreased expression of genes encoding for complement factors in line with the observed biological pathway activity patterns.<h4>Conclusion</h4>We present a comprehensive transcriptome analysis of explanted decellularized heart valve homografts providing insights into the biological processes leading to continued degeneration and ultimately loss of function. The degeneration of decellularized homografts is driven by inflammation, fibrosis, and extracellular matrix (ECM) remodelling, reinforced by ongoing oxidative stress rather than by a mainly complement-driven humoral immune response.

Extracellular Vesicles (EVs) have been increasingly recognized as mediators of intercellular communication in pancreatic tissues, carrying molecular cargo reflective of their cellular origin; however, their direct isolation from solid tissue remains technically challenging due to the high enzymatic activity, lipid content, and structural fragility of the pancreas. Here, we establish a pancreas-adapted and reproducible workflow for the isolation of EVs directly from human and mouse pancreatic tissues across tumor and non-tumorous contexts. This approach integrates controlled tissue processing, gentle enzymatic-mechanical dissociation, and sequential centrifugation to enable efficient vesicle recovery while preserving structural integrity and minimizing contamination. The isolated EVs exhibited characteristic size distributions, intact morphology, and enrichment of canonical EV markers, accompanied by depletion of intracellular components. Quantitative analyses demonstrated high reproducibility across independent isolations, with a coefficient of variation of less than 6%. Proteomic profiling revealed selective enrichment of vesicle-associated proteins and preservation of molecular differences between tumor and normal pancreatic tissues, indicating that tissue-derived EVs retain disease-associated protein signatures. Together, these results establish a standardized framework for pancreas-derived EV isolation that supports reproducible downstream analyses and facilitates molecular characterization of pancreatic tissues.

First-trimester prediction of preeclampsia (PE) remains a major clinical challenge, particularly outside specialized fetal medicine centers. This study aimed to identify and validate serum protein biomarkers for early PE prediction using an integrated proteomic approach. A prospective cohort of 64 first-trimester singleton pregnancies (32 future PE cases, 32 matched controls) was analyzed. Untargeted proteomics was performed using DIA-PASEF-MS, followed by targeted cross-platform verification with MRM-MS. Machine learning classifiers (support vector machines, SVM, and random forest) were trained on differentially abundant proteins (FDR < 0.01, VIP > 1.5). DIA-MS identified 33 protein markers associated with complement activation, IGF transport regulation, and platelet degranulation. An SVM model with a linear kernel achieved 95% accuracy (AUC = 0.95, sensitivity = 95%, specificity = 97%). Four markers (AFM, AHSG, C8A, IGHG1) were confirmed across platforms, confirming the discovery findings. Cross-platform correlation was high: 71% of overlapping proteins showed r > 0.5 (<i>p</i> < 0.001), with the highest concordance observed for potential PE marker AHSG (r = 0.8, <i>p</i> < 0.001). PRSS1, IGHV1-4, and SERPINC1 showed a strong correlation with proteinuria (|r| > 0.5, <i>p</i> < 0.05), linking the proteomic signature to clinical severity. Integrated DIA-MS and MRM-MS proteomics yields a reproducible, high-performance serum signature for first-trimester PE prediction. The identified markers reflect core pathophysiological pathways and offer potential to augment current FMF-based screening algorithms.

Patients with end-stage kidney disease (ESKD) receiving maintenance dialysis have a high burden of heart failure (HF), with cardiovascular disease the leading cause of death, yet are largely excluded from randomized trials. Using the TriNetX federated electronic health record network, we emulate a target trial comparing initiation of glucagon-like peptide-1 receptor agonists (GLP-1RAs) versus dipeptidyl peptidase-4 inhibitors (DPP-4is) in diabetic ESKD patients with HF, undergoing maintenance dialysis. We identify a primary ESKD-HF population and a confirmatory dialysis-dependence-coded HF subset, and apply a new-user, active-comparator design and use propensity score matching to balance baseline characteristics. In the primary ESKD-HF population, GLP-1RAs use is associated with a lower risk of the primary composite ischemic cardiovascular events plus HF exacerbations, than DPP-4i initiation (31.7% vs. 41.4%; HR 0.72[0.64-0.82], P < 0.0001), with concordance reductions in ischemic events (HR 0.74), HF exacerbations (HR 0.76), all-cause mortality (HR 0.68), and a death-inclusive composite (HR 0.72). Results are consistent in multivariable models, across prespecified subgroups, and in extensive sensitivity analyses, and are corroborated in the confirmatory dialysis-dependence-coded HF subset HR (0.71[0.60-0.85]). These findings provide real-world evidence suggesting GLP-1RAs may improve cardiovascular outcomes in dialysis-dependent diabetic ESKD with HF and support prospective randomized evaluation.

Solid tumors often evade TCR-engineered αβ T cells when antigen expression varies or when the restricting Human Leukocyte Antigen (HLA) allele is lost. γδ T cells, in contrast, detect cellular dysregulation through non-peptide/Major Histocompatibility Complex (MHC) cues, including phosphoantigens and stress ligands, and can be developed as allogeneic therapies. Although intratumoral γδ T cell signatures are associated with improved outcome across cancers, γδ recognition itself is broad and still selected within the thymus just as αβ T cell receptors (TCRs) are. It does not, however, anchor specificity to a defined driver-mutation pMHC epitope. We therefore asked whether a high-affinity, co-receptor-independent αβ TCR could graft oncogenic-driver specificity onto γδ T cells while leaving the endogenous γδ TCR intact. We knocked the KRASG12V/HLA-A*11:01 TCR A11v into primary human γδ T cells. Engineered cells co-expressed the transgenic αβ TCR and the endogenous γδ TCR and lysed KRASG12V/HLA-A*11:01⁺ tumor cells in vitro and in vivo. To cover potential resistance through loss of HLA-A*11:01, we delivered an mRNA lipid nanoparticle (LNP) encoding a secreted mesothelin×CD3 (M5) bispecific T cell engager (TCE). LNP-M5 produced circulating TCE that redirected γδ A11v T cells and polyclonal bystander T cells to kill mesothelin⁺ targets, accompanied by development of higher γδ A11v T cell counts in vivo. In humanized mice bearing mixed HLA-A*11:01⁺ and HLA-A*11:01 - KRASG12V tumors, γδ A11v T cells produced transient control, whereas adding LNP-M5 yielded complete responses and prolonged survival. Thus, this two-part therapy couples invariant driver targeting to tunable redirection and addresses loss of the restricting HLA allele, a central escape route for TCR-based therapy. It provides an off-the-shelf reagent to enable KRAS-anchored treatment with the ability to redeliver the reagent.

Midbrain dopamine neurons release dopamine not only from their axons but also from their somata and dendrites. Shared and distinct properties have been proposed for somatodendritic and axonal release, but the mechanisms of somatodendritic release remain unclear. We here used gene knock-out, electrophysiology, and imaging to define roles of the synaptic vesicle priming protein Munc13 in somatodendritic dopamine release in comparison with axonal secretion. We characterized mice of either sex and found that Munc13 ablation decreased evoked but not spontaneous somatodendritic dopamine transmission measured as D2 receptor-mediated currents. Imaging with a fluorescent sensor confirmed the importance of Munc13 in evoked somatodendritic and axonal dopamine secretion. Pharmacological experiments revealed a modest contribution of release from norepinephrine axons to D2 receptor-mediated currents, and the relative contribution was enhanced after Munc13 knock-out. Altogether, these data establish important roles of Munc13 in evoked somatodendritic release. These roles are similar to Munc13 functions in axonal dopamine release and at fast synapses. Spontaneous midbrain dopamine release was not impaired by Munc13 ablation from dopamine neurons and may rely on a release pathway that is independent of the prototypical release machinery employed at synapses.

Heparin, a polyanionic anticoagulant, is widely used clinically but requires neutralization to restore normal blood coagulation. Protamine sulfate (PS) is the standard antidote, yet limitations exist. Herein, a polycationic, cavity-containing pillar[6]arene macrocycle (P12+) demonstrates superior heparin neutralization in physiological media compared to its lower-valency analog P10+, and performance comparable to PS. MTT assays reveal low toxicity of the P12+, suggesting feasibility for in vivo use. Dynamic light scattering reveals that P12+ forms nanoscale complexes below 400 nm, smaller than PS-heparin aggregates. Remarkably, P12+ exhibits enhanced neutralization of low-molecular-weight heparin, a known limitation of PS. Additionally, heparin detection is achieved via a host-guest indicator displacement assay (IDA) using methyl orange (MO). UV-vis studies show ∼80% MO release with P12+, compared to ∼40% with P10+ in the presence of heparin. This response is detectable in undiluted human plasma for P12+, whereas P10+ shows a suitable response in 5% diluted plasma. A fluorescence-based IDA enables sensitive detection down to 0.11 U/mL, below the therapeutic range (0.80-2.00 U/mL). Molecular dynamics simulations support these findings, revealing that weaker host-guest interactions and favorable electrostatics in P12+ facilitate efficient MO release.

<h4>Background</h4>Human pluripotent stem cells (hPSCs) offer a promising source of hepatocytes for disease modeling and drug screening. However, hepatocytes derived in conventional two-dimensional (2D) cultures often exhibit incomplete maturation, limiting their physiological relevance. The molecular mechanisms underlying the improved differentiation observed in three-dimensional (3D) culture systems remain poorly understood.<h4>Results</h4>Induced PSCs (iPSCs) were differentiated into hepatic progenitors under 2D monolayer culture. At the hepatic progenitor stage, cells were either continued in 2D or re-aggregated into 3D organoids for hepatocyte maturation. Both conditions were cultured under identical experimental conditions throughout terminal differentiation for controlled comparison. At the end of differentiation, cultures were evaluated by immunostaining, gene and protein expression, functional assays, and RNA sequencing. Compared with 2D cultures, 3D hepatic organoids showed higher expression of mature hepatocyte markers (ALB, CPS1, AAT, and CYP3A4) and improved function, including increased albumin secretion, glycogen storage, and urea production. RNA sequencing identified 1,266 differentially expressed genes with upregulated genes enriched in hepatic metabolic pathways and downregulated genes associated with focal adhesion and extracellular (ECM)-receptor interaction.<h4>Conclusions</h4>These findings demonstrate that 3D organoid culture enhances iPSC-derived hepatocyte maturation by activating liver-specific transcriptional programs, while suppressing progenitor-associated gene signatures, supporting its use for liver disease modelling and drug metabolism studies.

HIV persistence within anatomical reservoirs remains the primary barrier to achieving an HIV cure. While antiretroviral therapy effectively suppresses plasma viremia, it does not eliminate integrated proviral genomes that persist in long-lived cellular compartments. The central nervous system (CNS) is a clinically important HIV reservoir, characterized by immune privilege and the persistence of tissue-resident infection despite effective antiretroviral therapy (ART). Evidence from postmortem studies reveals that HIV DNA, RNA, and even intact replication-competent proviruses remain detectable in brain tissue from virally suppressed people with HIV. Evidence derived primarily from in situ approaches and viable-cell studies supports myeloid-lineage reservoirs, particularly microglia and CNS-associated macrophages, as key cellular sources of persistence, while the extent and biological relevance of astrocyte infection remains debated. These reservoirs exhibit transcriptional activity and are associated with chronic neuroinflammation, which may contribute to HIV-associated neurocognitive disorders, despite systemic viral suppression. Here, we synthesize recent findings from autopsy brain studies, including work enabled by major biorepositories, such as the National NeuroHIV Tissue Consortium and rapid-autopsy programs, including the Last Gift, both of which are essential for studying HIV reservoirs in the CNS. We summarize methodologies for detecting and characterizing HIV in brain tissue, highlight heterogeneous patterns of regional distribution and compartmentalization, and review emerging links between CNS persistence and neuroinflammation. We conclude with priorities for harmonized tissue processing, multi-modal single-cell and spatial profiling, and coordinated cross-cohort analyses to clarify the contribution of CNS reservoirs to neuroHIV pathogenesis and systemic rebound.

TONSL safeguards genome stability by facilitating replication-dependent DNA damage repair and protecting stalled replication forks through homologous recombination. Mutations in TONSL cause SPONASTRIME dysplasia, a rare skeletal disorder. We reveal that TONSL homo-dimerizes via its ubiquitin-like domain (UBL), and two recurrent SPONASTRIME dysplasia causative variants (R934W and G973R) abolish this dimerization. Crystal structures at 1.9 Å resolution show UBL^WT forms domain-swapped dimers that assemble into ring-like octamers. The R934W variant eliminates critical hydrogen bonds and introduces steric clashes, forcing monomeric conformation. G973R destabilizes an evolutionarily conserved residue within a conformationally restricted β-turn. Biochemically, UBL^WT exists as dimers while UBL^R934W remains monomeric. Functionally, dimerization-deficient variants fail to suppress replication stress-induced DNA damage, show impaired RAD51 foci formation, and exhibit severely compromised survival following genotoxic stress. These findings establish TONSL dimerization as essential for genome maintenance and provide structural and mechanistic insights into SPONASTRIME dysplasia pathogenesis.

Microglia, the brain's innate immune cells, can adopt a wide variety of activation states relevant to health and disease. Dysregulation of microglial activation occurs in numerous brain disorders, and driving or inhibiting specific states could be therapeutic. To discover regulators of microglial activation states, we conducted CRISPR interference screens in induced pluripotent stem cell (iPSC)-derived microglia for inhibitors and activators of six microglial states. We characterized 31 regulators at the single-cell transcriptomic and cell-surface proteome level in two distinct iPSC-derived microglia models, uncovering protein markers of relevant states. We functionally characterized several multi-state regulators. ZNF532 and PRDM1 knockdown drive disease-associated, lipid-rich signatures and enhance phagocytosis while showing opposing effects on antigen-presentation signatures. DNMT1 knockdown results in widespread loss of DNA methylation, activating negative regulators of interferon signaling. These findings provide a framework to direct microglial activation to selectively enrich microglial activation states, define their functional outputs, and inform future therapies.

De novo variants in the ubiquitin-proteasome pathway are linked to autism spectrum disorder (ASD), yet their functional impact on neurodevelopment remains poorly understood. We investigated USP15, a deubiquitinating enzyme with rare damaging variants identified in individuals with ASD, using isogenic human iPSC-derived brain organoids and single-cell transcriptomics. USP15 mutant organoids showed genotype-dependent, progenitor-centered alterations during corticogenesis. Heterozygous organoids modeling haploinsufficiency displayed a shift toward later pseudotime states together with altered maturation and synaptic organization of deep-layer neurons. In contrast, homozygous organoids showed broader phenotypes, including mitotic suppression, aberrant HOX gene expression, and stress-response activation. Regulon analysis showed reduced activity of progenitor-associated regulons, including SOX2, NR2F1, and NR2F2, in heterozygous organoids, whereas homozygous organoids exhibited broader changes in transcriptional regulatory networks. Furthermore, USP15 mutant-associated gene expression patterns were significantly enriched for established ASD risk genes. Comparison with the mouse brain perturbation atlas showed that the transcriptional signature of the USP15 mutant showed notable overlap with those of Fezf2 and Foxp1 mutants, key regulators of deep-layer projection neuron identity. These findings characterize genotype-dependent neurodevelopmental phenotypes associated with reduced USP15 dosage and provide a human neural framework for investigating ASD-relevant developmental mechanisms in the context of a rare ubiquitin-pathway variant.

Epilepsy is a prevalent neurological disease, with one-third of individuals becoming nonresponsive to antiepileptic drugs and developing drug-refractory epilepsy (DRE). Here we identify activation of cyclic GMP-AMP synthase (cGAS), a double-stranded DNA sensor that induces type I interferon (IFN) signaling, in human DRE brain tissue. Microglia from individuals with DRE exhibit a robust type I IFN signature and the activation of upstream cGAS-STING signaling. Further, in mouse models of Dravet syndrome, a genetic form of DRE, we similarly detect activation of the cGAS pathway. We show that microglial cGAS can be activated by DNA released from hyperexcitable neurons. Genetic reduction and pharmacological inhibition of cGAS attenuates seizure phenotypes, reduces glial inflammatory signatures and normalizes neuronal transcriptomic changes in mice with Dravet syndrome. Together, these findings identify cGAS-mediated neuroimmune signaling as a contributor to seizure pathology in Dravet syndrome and highlight this pathway as a potential therapeutic target.

Huntington's disease (HD) is a fatal neurodegenerative disorder caused by an expanded CAG repeat within exon 1 of the huntingtin (HTT) gene, resulting in a mutant protein that drives neuronal dysfunction and loss. A key event in the pathogenesis of HD is proteolytic cleavage of mutant HTT, which generates aggregation-prone N-terminal fragments that contribute to toxicity. Strategies that prevent this process thus hold therapeutic potential. Here we develop CRISPR base editors that generate proteolysis-resistant HTT isoforms by disrupting the splice acceptor of HTT exon 13, an exon that encodes critical proteolytic cleavage sites implicated in N-terminal fragment production. When delivered to the striatum of an HD rodent model, these editors reduced HTT fragment formation, decreased aggregation, improved functional deficits and attenuated brain atrophy. Collectively, these results demonstrate the potential of base editing and splice-site modulation to mitigate mutant HTT toxicity in HD.

Inhibition of androgen receptor (AR) signaling remains the cornerstone of systemic therapy for advanced prostate cancer (PC). However, a subset of aggressive tumors either arises de novo with neuroendocrine features or emerges under treatment pressure through lineage plasticity and AR independence. These lethal states are encompassed within the spectrum of aggressive-variant prostate cancer (AVPC), an umbrella term that includes both histologically confirmed neuroendocrine prostate cancer (NEPC)-comprising de novo NEPC and treatment-emergent NEPC (t-NEPC)-and clinically or molecularly defined AVPC lacking histologic confirmation but sharing neuroendocrine-like, AR-indifferent, or small-cell features. These phenotypes are characterized by rapid progression, visceral dissemination, low or discordant prostate-specific antigen (PSA) levels relative to tumor burden, and poor prognosis. Treatment options for NEPC/AVPC remain limited and largely rely on platinum-based chemotherapy, which usually provides only modest and transient benefit. This unmet need has intensified interest in lineage-associated vulnerabilities. Delta-like ligand 3 (DLL3), an inhibitory Notch ligand with restricted expression in normal adult tissues, is aberrantly upregulated in several neuroendocrine malignancies and has emerged as a clinically actionable target. In prostate cancer, DLL3 expression is enriched in neuroendocrine tumor cells, being detected in approximately 76.6% of castration-resistant NEPC compared with only 12.5% of castration-resistant adenocarcinoma, supporting its development as both a biomarker and therapeutic vulnerability. Clinical success of DLL3-targeted therapies in small-cell lung cancer further supports evaluation of DLL3-directed strategies in NEPC and related AVPC states. This review summarizes the biological rationale, translational evidence, and emerging clinical data supporting DLL3-targeted therapies in prostate cancer. Investigational platforms include antibody-drug conjugates, bispecific and trispecific T-cell engagers, and DLL3-directed radiopharmaceuticals. Early clinical studies suggest that activity is largely confined to DLL3-expressing neuroendocrine tumors, highlighting the importance of biomarker-guided patient selection. Delta-like ligand 3-directed therapies may reshape the management of DLL3-expressing prostate cancer if ongoing efforts to refine biomarkers improve patient enrichment and optimize trial design are successfully translated into clinical practice.

Herein, divalent cations such as Ca²⁺, Sr²⁺, and Mg²⁺ are widely studied for their contribution to the bioactivity and performance enhancement of biomaterials intended for bone applications. The incorporation of functional ions into these bioceramics thus represents a promising and economically viable approach. In contrast, Fe, Al, and B cations, although recognized for their influence on structure, chemical stability, and certain physicochemical properties, remain relatively unexplored in relation to the bioactivity of bone-related biomaterials. The present study focuses on the development of three bioactive bioceramics, successfully synthesized by the conventional solid-state reaction method, followed by heat treatments between 100 °C and 1000 °C. These bioceramics are based on the dicalcium silicate phase (Ca₂SiO₄), into which bioactive trivalent cations, such as Boron oxide (B₂O₃), Alumina (Al₂O₃) and Hematite (Fe₂O₃) have been incorporated, named CSF, CSA, CSB respectively. Furthermore, the bioactivity of these bioceramics were evaluated by immersing them in simulated body fluid (SBF) and artificial saliva (AS) for 24 hours at 37 °C. The samples were then characterized by powder X-ray diffraction (XRD), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), Fourier transform infrared spectroscopy (FT-IR), and UV spectroscopy. These analyses were complemented by using transmission electron microscopy (TEM). X-ray diffraction (XRD) results confirmed that the Dicalcium silicate (Ca₂SiO₄) phase is predominant in the three bioceramics, with secondary bioactive phases such as Grossite (CaAl₄O₇), Tricalcium borate (Ca₃B₂O₆) and Brownmillerite (Ca₂Fe₂O₅), specific to each incorporated oxide. The optical band gap of the samples, determined by UV-visible spectrophotometry, ranged from 3.60, 3.21 and 2.99 eV for CSF, CSB, CSA respectively. Antibacterial activity, evaluated against Gram-negative (<i>Pseudomonas aeruginosa</i>) and Gram-positive (<i>Staphylococcus aureus</i> and <i>Enterococcus faecalis</i>) bacteria, revealed significant inhibition of <i>Staphylococcus aureus</i>. XRD analyses after only 24 hours immersion in simulated media, complemented by scanning electron microscopy (SEM) observations, showed that these bioceramics rapidly promote the formation of a bone-like hydroxyapatite (Ca₁₀(PO₄)₆(OH)₂) layer, demonstrating their potential as biomaterials for bone regeneration. The results obtained indicate that the incorporation of bioactive oxides into dicalcium silicate may significantly improve its properties, thus enhancing its potential for tissue regeneration applications.

Cell-to-cell signaling between niche and stem cells regulates tissue renewal. While the identity of many mediating factors is known, it is largely unknown whether stem cells optimize their receptiveness to niche signals according to the niche organization. Here, we show that Lgr5+ small intestinal stem cells (ISCs) regulate the morphology and orientation of their secretory apparatus to match the niche architecture, and to increase transport efficiency of niche signal receptors. ISCs orient their Golgi apparatus laterally towards Paneth cells of the epithelial niche, and divide Golgi into multiple stacks. Stem cells with multiple lateral Golgi transport stem cell receptors with a higher efficiency than cells with one single Golgi. The lateral Golgi orientation and enhanced receptor transport requires A-kinase anchor protein 9 (Akap9), and is necessary for normal renewal capacity. Moreover, reduced Akap9 in aged ISCs renders ISCs insensitive to niche-dependent modulation of Golgi stack number and transport efficiency. Our results reveal a stem cell-specific Golgi complex configuration that facilitates efficient niche signal reception and tissue renewal, which is compromised in the aged epithelium.

<h4>Background</h4>Hepatocellular carcinoma (HCC) has a poor prognosis with lymph node metastasis (LNM), severely impacting survival. Protein post-translational modifications (PTMs) regulate tumor microenvironment (TME) dynamics, but their role in HCC LNM is unknown.<h4>Methods</h4>This study systematically analyzed single-cell RNA sequencing (scRNA-seq) and bulk transcriptomic data from TCGA-LIHC and validation cohorts. Key cellular subpopulations and differentially expressed genes (DEGs) were identified. PTM-related candidate genes were identified through intersection analysis. Prognostic genes were determined via Cox regression and integrated into a random survival forest (RSF) model. Patients were stratified to assess associations with TME remodeling, genomic alterations, immune checkpoints, and drug sensitivity. Pseudotime trajectory and transcription factor activity analyses elucidated molecular dynamics.<h4>Results</h4>Hepatocyte populations were significantly expanded in both LNM and non-LNM HCC groups. The intersection of scRNA-seq DEGs, bulk DEGs, and PTM-related genes identified five candidate genes, among which PJA1 and RFPL4B identified as potential prognostic risk factors and were incorporated into a robust RSF model. High-risk patients exhibited poor survival outcomes, increased tumor mutational burden, and pronounced tumor microenvironment alterations. PJA1 expression was negatively correlated with hepatocyte infiltration. Pseudotime analysis revealed that LNM-associated hepatocytes were arrested in early differentiation states, coinciding with dysregulated transcription factor activity.<h4>Conclusion</h4>This study computationally identified PJA1 and RFPL4B as post-translational modification-related candidate prognostic genes potentially associated with HCC LNM. The proposed risk model stratified patients into subgroups characterized by distinct tumor microenvironment features and genomic instability. Computational analysis suggested that arrest of hepatocyte differentiation and dysregulation of transcription factor networks may be implicated in metastatic progression, thereby offering novel therapeutic insights.

Accumulating evidence highlights the pivotal role of ferroptosis in diverse cardiovascular diseases (CVDs) such as atherosclerosis, aortic aneurysm, aortic dissection, vascular aging, calcific aortic valve disease, myocardial ischemia reperfusion injury, cardiomyopathy, and heart failure. Studies on the molecular mechanism of ferroptosis have revealed potential targets for the treatment of CVDs, and a large number of studies have reported evidence of drug-mediated ferroptosis and iron metabolism mechanisms intervening in CVDs. This review delves into the latest molecular characteristics of ferroptosis, associated biological processes, and regulatory pathways. Notably, various compounds like iron chelators (e.g., deferoxamine, deferiprone), chloroiodohydroxyquin and its derivatives, antioxidants (e.g., vitamin E, lipoic acid, selenium), Ferrostatin-1 (Fer-1), etc., have demonstrated efficacy in animal studies against CVDs, offering cardiovascular protection. In summary, this review outlines the role of ferroptosis in CVDs, the regulatory effects of relevant drugs (including natural products) on ferroptosis within cardiovascular contexts, aiming to provide valuable insights for CVDs management and the development of novel therapeutics.

Mitochondrial diseases are a heterogeneous group of inherited disorders caused by defects in the mitochondrial genome or nuclear genes encoding proteins essential for mitochondrial function. These conditions are characterised by progressive dysfunction of tissues with high energy demands, particularly the nervous and muscular systems. In recent years, increasing consideration has been paid to the role of autophagy-the cellular mechanism for the degradation and recycling of intracellular components in the pathogenesis of mitochondrial diseases. This review synthesizes current knowledge on molecular mechanisms of macroautophagy and selective forms of au-tophagy targeting specific organelles and structures: mitophagy, pexophagy, ribophagy, ER-phagy, aggrephagy, lipophagy, lisophagy, and nucleophagy. Using classic mitochondrial syndromes-Kearns-Sayre syndrome (KSS), MERRF, and MELAS, as well as various mitochondrial myopathies-as examples, we discuss experimental evidence indicating both compensatory activation of autophagy and its insufficiency or blockade at different stages. Furthermore, we examine the link between deficiencies of key fatty acid β-oxidation enzymes (VLCAD, MCAD, CPT2) and impaired autophagic flux, including secondary defects of mitophagy mediated by energy deficiency. The review systematises current understanding of how dysregulation of selective autophagy promotes the accumulation of damaged mitochondria, oxidative stress, inflammation, and cell death in mitochondrial diseases. Prospects for therapeutic modulation of autophagy as a potential approach to treating these disorders are discussed.

Understanding whether long-term vascular cambium vitality in ancient trees reflects progressive decline or adaptive reprogramming is central to grasping woody plant longevity. We performed an integrative analysis of functional traits, transcriptome profiles, and metabolomic landscapes of cambial zone enriched from 80-, 500-, and 1000-year-old <i>Styphnolobium japonicum</i> trees. With increasing tree age, the vascular cambium showed fewer cell layers, reduced thickness, and lower auxin, gibberellin, and IAA/ABA ratios, whereas bark thickness, malondialdehyde, abscisic acid, jasmonic acid, and salicylic acid contents increased. Transcriptomic and metabolomic analyses revealed that differentially expressed genes and metabolites were primarily enriched in the cell cycle, phytohormone signaling, and phenylpropanoid biosynthesis pathways. Specifically, genes associated with cell division were down-regulated in millennial trees, whereas phenolic acids, flavonoids, and lignin-related metabolites significantly accumulated. Piecewise structural equation modeling suggested associations among tree age, transcription factors, structural genes, metabolites, and cambial functional traits. These results indicate that cambial senescence is not a simple linear decay but a highly coordinated remodeling process, providing crucial evidence for delayed senescence in long-lived woody species.

Autophagy-lysosome dysfunction accompanies dermal fibroblast ageing, yet whether remodelling is transcriptional or post-transcriptional in primary human cells remains unresolved. We reanalysed the Genetic and Epigenetic Signatures of Translational Ageing Laboratory Testing(GESTALT) paired RNA sequencing (RNA-seq) and tandem mass tag (TMT) proteome from 82 donors (aged 22-89) using Data Integration Analysis for Biomarker discovery using Latent cOmponents (DIABLO) for supervised multi-omics integration, weighted gene co-expression network analysis (WGCNA), external Genotype-Tissue Expression(GTEx) transcriptomic comparison, network medicine proximity mapping and CDOCKER molecular docking. Three analyses converged on the autophagy-lysosome axis: Kyoto Encyclopaedia of Genes and Genomes (KEGG) Lysosome ranked first in discordant-quadrant analysis; gene set enrichment analysis (GSEA) identified vacuole organisation and macroautophagy as the top age-upregulated Gene Ontology (GO) terms; and WGCNA recovered KEGG Lysosome in the brown module. Module regression localised most proteomic age signals to the lysosomal degradative-capacity module, whereas the proteasome was unaffected. McNemar testing and GTEx comparison supported a protein-side, post-transcriptional origin. TCIRG1, CTSA and ATP6V0D1 were recurrent hubs. Network proximity computationally prioritised hydroxytyrosol as a lysosomal-degradative-capacity-preferential candidate, and CDOCKER on cathepsin A linked its advantage over tyrosol to an ortho-hydroxyl group forming additional hydrogen bonds. These results support protein-layer-dominant autophagy-lysosome remodelling as a feature of dermal fibroblast ageing and suggest a cell-type-resolved computational route from ageing proteomics to testable dietary candidates.

<h4>Background/aims</h4>Optimal pre-transplant chemotherapy exposure for pediatric patients with high-risk acute myeloid leukemia (AML) in first complete remission (CR1) remains undefined. Many patients achieve measurable residual disease (MRD)-negative remission early, raising the question of whether additional chemotherapy prior to allogeneic hematopoietic cell transplantation (HCT) improves outcomes. We evaluated the impact of pre-HCT chemotherapy cycles on post-transplant outcomes.<h4>Methods</h4>We conducted a retrospective single-center cohort study of pediatric patients (age 0-18 years) with high-risk AML undergoing first myeloablative allogeneic HCT in MRD-negative CR1 between 2011 and 2023. Patients were stratified by receipt of 1-2 versus 3-4 pre-HCT chemotherapy cycles. Patients in the 3-4 cycle cohort who had achieved MRD-negative remission by the end of cycle 2 were assessed and compared to the 1-2 cycle cohort for rates of two-year relapse-free survival (RFS) and cumulative incidence of relapse. All patients in both cohorts were assessed for rates of two-year overall survival (OS) and one-year non-relapse mortality (NRM).<h4>Results</h4>Forty patients met inclusion criteria; 22 (55%) received 1-2 cycles and 18 (45%) received 3-4 cycles. Excluding patients who were not MRD-negative after 2 cycles, two-year RFS was 66% (95% CI 32-86) for 3-4 cycles vs 68% (45-83) for 1-2 cycles (Hazard Ratio [HR] 0.88 [95% CI 0.26-3.01], p=0.84). The cumulative incidence of relapse at two years was 34% (9-62) in the 3-4 cycle cohort and 14% (3-31) in the 1-2 cycle cohort (HR 2.39 [0.57-10.1], p=0.23). Two-year OS was 77% (49-91) for 3-4 cycles vs 67% (46-84) in the 1-2 cycle cohort (HR 0.63 [0.18-2.14], p=0.46). One-year NRM was 0% in the 3-4 cycle group and 18% (5-37) in the 1-2 cohort (HR NE, p=0.06).<h4>Conclusions</h4>Among pediatric patients with high-risk AML undergoing allogeneic HCT in MRD-negative CR1 at a single center, 3-4 pre-transplant chemotherapy cycles were not associated with statistically significant differences in overall or relapse-free survival compared to 1-2 cycles, although the study was underpowered to exclude clinically meaningful differences. Consolidation with HCT after 1-2 cycles of chemotherapy, once MRD-negative remission is achieved, warrants prospective exploration of risk- and remission-guided treatment strategies.

<h4>Background</h4>Cardiovascular diseases (CVDs) remain the leading cause of mortality worldwide, with inflammation central to disease progression. Emerging evidence reveals intricate crosstalk between metabolic reprogramming and epigenetic modifications in cardiovascular inflammatory injury, offering novel therapeutic strategies.<h4>Objective</h4>This review elucidates the regulatory mechanisms underlying the metabolic-epigenetic axis in cardiovascular inflammation and explores both conventional and traditional Chinese medicine (TCM)-based intervention strategies.<h4>Results</h4>Three findings are highlighted: (i) the succinate-hypoxia-inducible factor 1α (HIF-1α)-interleukin-1β (IL-1β) and itaconate-nuclear factor erythroid 2-related factor 2 (Nrf2) axes form an opposing pair defining macrophage polarization; (ii) acetyl-CoA, α-ketoglutarate (α-KG), lactate, and β-hydroxybutyrate (β-OHB) couple metabolism to histone modifications (H3K27ac, H3K27me3, H3K18la, H3K9ac) and DNA 5-hydroxymethylcytosine (5hmC), establishing trained immunity; (iii) mitochondrial dysfunction (mtDNA-cGAS-STING; mitochondrial ROS-NLRP3 inflammasome) and ferroptosis (GPX4/ACSL4) integrate these signals at the organelle level. TCM compounds (berberine, Shengmai Injection, tanshinone IIA, ginsenosides) modulate this axis at multiple nodes, with clinical support from the Phase III China Tongxinluo Study in Acute Myocardial Infarction (CTS-AMI) trial.<h4>Conclusions</h4>The metabolic-epigenetic axis offers a tractable framework for precision anti-inflammatory cardiovascular therapy. Mechanistic studies and biomarker-guided trials are required to validate combined TCM-based interventions.

Natural killer (NK) cells are uniquely equipped to eliminate transformed cells without prior antigen sensitization, yet their therapeutic potential in solid tumors remains only partially realized. At the center of this paradox lies a complex network of inhibitory pathways, dominated by killer cell immunoglobulin-like receptors (KIRs) and the CD94/NKG2A axis, which continuously calibrate NK-cell self-tolerance and effector competence. Tumors exploit these regulatory circuits through dynamic remodeling of HLA-I expression: while loss of classical HLA-I impairs CD8⁺ T-cell recognition, preservation or upregulation of non-classical HLA, particularly HLA-E, sustains inhibitory signaling and promotes immune escape. These mechanisms are further amplified by the tumor microenvironment (TME), where stromal barriers, hypoxia, metabolic stress, and immunosuppressive networks collectively restrict NK-cell infiltration, persistence, and cytotoxicity. Such multilayered suppression helps explain why therapeutic blockade of KIR or NKG2A alone has yielded only modest clinical benefit in most solid tumors, despite compelling biological rationale. Emerging evidence suggests that the KIR- and CD94/NKG2-centered network should be viewed not only as a therapeutic target but also as a framework for the next-generation of NK-cell-based immunotherapies. Future strategies will likely combine checkpoint modulation with donor- and patient-tailored NK-cell selection, engineered NK-cell products with enhanced metabolic resilience and reduced checkpoint sensitivity, and interventions aimed at remodeling the tumor niche to restore trafficking, persistence, and functional fitness. In this mini-review, we discuss how KIR- and CD94/NKG2-mediated signaling is shaped by the TME and examine emerging combinatorial and personalized approaches designed to unlock the full therapeutic potential of NK cells in solid tumors.

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder defined by progressive motor neuron loss and TDP-43 proteinopathy, yet the upstream drivers of this pathology remain unclear. Oxidized phosphatidylcholines (PC-OxPL) have emerged as potent inducers of proteinopathy in the central nervous system (CNS), but their role in ALS has not been systematically explored. We identify a distinct PC-OxPL signature in the cerebrospinal fluid (CSF) of patients with sporadic ALS (sALS) and show that apolipoprotein E (apoE)-containing particles are the primary PC-OxPL carriers in this compartment. In human iPSC-derived motor neurons, PC-OxPL exposure triggered disease-relevant transcriptional alterations and TDP-43 pathology, establishing PC-OxPL as a mediator of ALS-like neurodegeneration <i>in vitro</i>. To counteract this toxicity, we engineered an Adeno-Associated Virus (AAV)-delivered single-chain antibody fragment (scFv), PC-OxPL-VecTab, targeting PC-OxPL neoepitopes. PC-OxPL-VecTab neutralized PC-OxPL-induced neurotoxicity, reduced TDP-43 aggregation, and prevented motor neuron death and behavioral deficits in a sALS CSF transfer mouse model. Intrathecal delivery of PC-OxPL-VecTab in minipigs achieved broad CNS biodistribution and transgene expression, supporting the feasibility of CNS delivery. These findings position PC-OxPL as a mechanistic contributor to ALS pathogenesis and establish PC-OxPL-VecTab as a therapeutic strategy for ALS with potential broader applicability to disorders associated with PC-OxPL accumulation.

Neurodegenerative diseases are targets for pridopidine therapy, which aims to improve quality of life through neuroprotective mechanisms that involve sigma-1 receptor (S1R) activation. Neurodegenerative motor and cognitive diseases are influenced by dopamine imbalance, where disruptions in pathways contribute to states that are hyperkinetic or hypokinetic, while current dopaminergic treatments are symptomatic rather than disease-modifying, especially for Huntington's disease and Amyotrophic lateral sclerosis. This review summarizes the mechanisms underlying pridopidine-mediated neuroprotection and examines the current evidence supporting its therapeutic potential. The S1R is an endoplasmic reticulum-mitochondria-associated chaperone involved in homeostasis of calcium, stress regulation, and mitochondrial function. Pridopidine is a small lipophilic molecule that crosses the blood-brain barrier and acts as an S1R agonist, with minimal dopamine D2 receptor occupancy. Activation of S1R by pridopidine modulates calcium signaling and enhances anti-apoptotic activity. Collectively, available evidence suggests that pridopidine may improve motor outcomes and slow disease progression in Huntington's disease and amyotrophic lateral sclerosis, supporting its promise as a disease-modifying therapeutic strategy.

Autism spectrum disorder (ASD) is a heterogeneous neurodevelopmental condition characterized by impairments in social communication, restricted and repetitive behaviors, sensory dysregulation, and frequent psychiatric comorbidities. Increasing attention has been directed toward acupuncture as a complementary neuro-modulatory intervention; however, its underlying molecular mechanisms remain incompletely understood. This review synthesizes evidence from preclinical, clinical, and molecular studies published between 2015 and 2025 to examine how acupuncture influences neurobiological pathways relevant to ASD. Current evidence indicates that acupuncture modulates multiple neurotransmitter systems, including glutamatergic, GABAergic, dopaminergic, serotonergic, and noradrenergic signaling, while also influencing neurotrophin-mediated plasticity, neuroinflammatory responses, and synaptic function. Studies conducted directly in ASD models demonstrate regulation of excitatory/inhibitory balance, monoaminergic signaling, neurotrophin pathways, and ASD-associated behavioral outcomes, whereas evidence from related neuropsychiatric conditions provides complementary mechanistic support for these pathways. Collectively, the findings suggest that acupuncture may act through coordinated modulation of interconnected neurotransmitter and neuroplasticity networks rather than a single molecular target. However, direct mechanistic evidence in ASD-specific models and clinical populations remains limited, and considerable heterogeneity exists among acupuncture protocols and outcome measures. Future studies integrating standardized stimulation paradigms with molecular, electrophysiological, neuroimaging, and behavioral assessments will be essential to validate the proposed mechanisms and clarify the translational potential of acupuncture in ASD.

Brucellosis remains among the most widespread global zoonoses, yet its genetic architecture-spanning host susceptibility and pathogen virulence-is increasingly being investigated. Advances in whole-genome sequencing, transposon mutagenesis, CRISPR screening, and machine learning-driven transcriptomics have accelerated marker discovery on both sides of the host-pathogen interface, although most signatures remain at discovery or early-validation stages. In hosts, polymorphisms in innate immune receptors (<i>TLR2</i>, <i>TLR5</i>, <i>VDR</i>), cytokine regulators (<i>TNF</i>, <i>CTLA4</i>), and novel candidates such as <i>ASAP1</i> are linked to differential susceptibility in humans and livestock. In <i>Brucella</i>, functional genomics shows that virulence relies on interconnected regulatory circuits that control the type IV secretion system, lipopolysaccharide biosynthesis, and stress responses, with gene essentiality shifting according to the host immune context. Antimicrobial resistance may involve unconventional mechanisms, including metabolic and regulatory adaptation, that are not fully explained by classical resistance-gene paradigms and may implicate metabolic reprogramming and ribosomal modulation. This review synthesizes these converging lines of evidence, highlights key gaps-chiefly the scarcity of functional validation for host associations and limited multi-omics integration-and proposes a roadmap for translating genetic markers toward future diagnostic refinement and therapeutic target discovery, urging a shift from descriptive cataloging toward mechanistic and clinical validation.

Huntington’s disease (HD) is a life-altering genetic neurodegenerative disorder, with cognitive, motor, and psycho-social effects that have consequential impacts on the individuals and their families. While current treatments improve disease symptoms, there are no FDA-approved therapies that prevent disease progression. Converging lines of evidence from human GWAS and mouse models point to DNA repair and handling (R/H) proteins as promising therapeutic targets due to their ability to modulate somatic expansion of the CAG repeat of HTT. The roles of DNA R/H HD modulator proteins are incompletely understood, in part, due to their relatively low cellular abundance and technical challenges in quantification. Here, we developed and validated targeted mass spectrometry assays quantifying DNA R/H proteins, spanning functions in mismatch repair, Fanconi anemia, and transcriptional regulation, using complementary workflows for timsTOF and Orbitrap platforms. We built species-specific experiment spectral libraries that outperformed in silico libraries for target detection. Applying this pipeline to an HTT-Q140 knock-in mouse HD model, we observed that DNA R/H protein abundances were largely unchanged in HD mice, while HTT and HAP40 showed increased nuclear association with disease progression. To facilitate translational research applications, we further developed a stable isotope dilution assay for absolute quantification of 11 human mismatch repair-associated proteins and generated an HTT knock-out human neuroblastoma cell line. Additionally, we used thermal proximity coaggregation profiling to characterize the endogenous interactomes of MMR proteins. We observed that HTT KO caused proteome down-regulation in selected DNA R/H proteins and reshaped the MMR protein interactome, with the most pronounced changes observed for MLH1 and PMS1 interactions. Overall, we established a validated, transferable assay for quantifying DNA R/H proteins in perturbation studies using human and mouse HD model systems and provide evidence that HTT influences the abundance and interaction landscape of proteins central to CAG repeat instability.

<h4>ABSTRACT</h4> Somatic instability (SI) of expanded DNA repeats is a hallmark of repeat expansion disorder (REDs) and drives onset and progression in Huntington’s disease (HD) yet the absence of target engagement (TE) biomarkers for SI-modulating therapies represents a critical gap to clinical development. Here, we describe the development of the unscheduled repair synthesis assay (URSA)—combining 5-ethynyl-2’-deoxyuridine (EdU) pulse-labeling with digital PCR or sequencing—and show that the CAG-expanded huntingtin ( HTT ) exon 1 allele is a highly active DNA repair hotspot in cells from people with HD (PwHD). Repair activity increases with repeat length, is allele-specific, and depends strongly on MSH3, a central driver of somatic expansion. URSA robustly quantifies MSH3 modulation in preclinical models within days compared to weeks or months required by conventional repeat-length measurements. Critically, substantial repair activity is detectable in peripheral blood mononuclear cells (PBMCs) from PwHD, where signal correlates with CAG length and improves predictive models of somatic expansion (SE) beyond age and CAG length alone. Unlike repeat-length changes, which require years to accumulate in blood, URSA signal is measurable within days. These findings establish DNA repair activity at the mutant HTT locus as a mechanistically grounded pharmacodynamic biomarker, enabling TE monitoring on a clinically actionable timescale, and with broad applicability to REDs and other diseases where modulation of the DNA damage response is therapeutically targeted.

Multiple therapeutic strategies are being developed to slow Huntington’s disease (HD) progression through targeted reduction of huntingtin (HTT) protein or mRNA. Despite HTT’s discovery over 30 years ago, its cellular functions remain incompletely understood, and the long-term consequences of HTT-lowering therapies remain unclear. We previously demonstrated that hepatic HTT loss in mice disrupts hepatocyte zonation and metabolism. Here, we investigate the physiological consequences of hepatic Htt loss. Across multiple models of Htt loss—including ubiquitous and hepatocyte-specific genetic knockouts and a therapeutically relevant Htt -targeting siRNA—there was elevated expression of IL-6/STAT3-driven acute phase response genes. Single-nucleus RNA sequencing reveals a zonal pattern of hepatocyte stress, most highly upregulated in pericentral hepatocytes, and identifies a distinct pericentral cluster of stressed hepatocytes that was enriched ∼9.6-fold following Htt knockout. Histological examination reveals that Htt loss results in increased hepatic pathology, including hepatic intranuclear inclusions, apoptosis, and necrosis, as well as prevalence of granulomas. Transcriptomic analysis reveals significant upregulation of metallothionein genes following Htt loss, as confirmed by elevated plasma metallothionein-1 (MT1) levels in knockout mice. These findings underscore important safety considerations for HTT-lowering therapies and suggest candidate biomarkers for monitoring hepatic off-target effects in clinical trials.

<h4>ABSTRACT</h4> Foundation models trained on electronic healthcare records (EHRs) have gained traction with the aim to transform personalised medicine. However, their interpretability is bound to redescribing the records the models were trained on, missing implicitly learned concepts and biases. Here, we show that human genetics provides an orthogonal layer to surface implicitly learned biological concepts and otherwise hidden risk factors. Re-implementing the generative transformer Delphi-2M in >500,000 UK Biobank participants, we performed genome-wide association testing on its 120 learned embeddings and identified 434 genome-wide-significant signals across 151 independent loci and 98 embeddings, revealing a heritable structure that feature-attribution methods cannot recover. Effector-gene mapping implicated cholesterol metabolism and an IL-1-family epithelial-alarmin pathway, supported by strong (>50-fold) enrichment for variants previously associated with blood lipids, body-mass index, and asthma. Loci recovered the targets of essentially all approved lipid-lowering and severe-asthma therapies, and another twelve drugs not obvious from genetic results based on single ICD-10 GWAS. Yet, embeddings poorly explained variation in pleiotropic risk factors, while still retaining most of their predictive value. Substantial improvements in predictive performance were hence confined to a minority of common diseases by adding specific diagnostic or organ-derived markers. Our findings suggest that human genetics might be most powerful as an orthogonal explanatory or regularising layer to train the next generation of EHR-based foundation models that likely benefit most from the addition of targeted biomarkers to advance personalised medicine.

<h4>Background</h4>The genetic dissection of complex traits in livestock continues to pose a significant challenge in the field of animal genetics and breeding. Although traditional genome-wide association studies (GWAS) are capable of localizing genetic variants associated with specific traits, they are insufficient to elucidate the underlying physiological mechanisms. An integrated analysis of multi-trait GWAS and multi-transcriptomic data systematically identifies key tissues and cell types influencing complex traits in beef cattle and elucidates their genetic regulatory basis.<h4>Results</h4>We systematically mapped tissue- and cell-type-specific regulatory architectures underlying 20 economically important traits in beef cattle. Tissue-level analyses revealed distinct trait-tissue associations: fatty acid traits, including C16:0 and C20:4, were enriched in liver; carcass traits, including marbling score and carcass weight, in renal cortex/medulla and longissimus dorsi muscle; meat-quality traits such as pH in cartilaginous tissues; and total fat content in bone marrow. At cellular resolution, analysis of eight trait-associated tissues identified 38 discrete cell types. Myofibers were significantly associated with most carcass traits, including rib-eye area and backfat thickness, whereas hepatocytes emerged as key regulators of fatty acid and meat-quality traits, such as C16:0 and crude protein content. Transcription factor analysis identified cell-type-specific regulators: TBX15, SOX6, and TCF12 in myofibers; FOXA2 and NR1H4 in hepatocytes; and IRF8 and IKZF1 in microglia. Notably, hepatocytes and microglia showed complementary, trait-specific association patterns: hepatocytes were enriched for C16:0 associated saturated fatty-acid metabolic pathways, while microglia were enriched for C16:1 and unsaturated fatty-acid-related pathways, suggesting potential cross-tissue coordination in lipid regulation.<h4>Conclusions</h4>Our study links specific tissues and cell types to phenotypic variation in beef cattle and identifies core transcriptional regulators and pathways driving trait variation. These cell-resolved maps provide mechanistic insight into how genetic variation shapes economically important traits, offering a valuable resource for functional studies, cell-informed precision breeding strategies, and the design of large-scale molecular phenotyping.

With the progressive shift of minimally invasive aesthetic medicine toward regeneration-oriented therapeutic paradigms, injectable materials have evolved from providing simple volumetric augmentation to enabling tissue remodeling and long-term structural improvement. Regenerative functional microspheres, as representative biostimulatory injectable materials have demonstrated expanding clinical value in facial rejuvenation, skin texture enhancement, and scar repair. However, studies on different material systems remain fragmented with respect to their mechanisms of action, manufacturing strategies, and clinical outcomes, and the lack of systematic integration and cross-comparison has hindered manufacturing standardization and standardized clinical application. Focusing on aesthetic medical applications, this review systematically summarizes the regenerative mechanisms, major material systems, fabrication strategies, and clinical evidence of regenerative functional microspheres. Centered on the critical clinical effects of biostimulation and collagen remodeling, this review highlights recent advances in regenerative mechanisms and material design, outlines the influence of key fabrication technologies on product uniformity and safety, and systematically analyzes major clinical indications and therapeutic characteristics, while also discussing key challenges and future development trends in this field. This review aims to provide a structured overview to guide material design, process optimization, and clinical translation of regenerative microspheres, offering a reference framework for innovation and application of regenerative aesthetic materials.

Defining cell classes is central to the analysis of growing single-cell RNA sequencing (scRNA-seq) atlases. Marker genes are most often identified by differential expression (DE) methods that assess genes one at a time, ignoring the redundancy and complementarity revealed when genes are considered jointly. Working with binarized expression data, we instead seek discriminating <i>panels</i> of genes that together are specific to a cell type, framing marker-panel selection as a variant of the minimal set-covering problem in combinatorial optimization. This formulation efficiently searches the vast space of candidate panels, exploits the large cell numbers typical of scRNA-seq, and is robust to zero-inflation. Using blood and brain data, we show that our method, CellCover, reduces gene redundancy and captures cell-class-specific signals distinct from those found by DE. Transfer-learning experiments across mouse, primate, and human data demonstrate that CellCover identifies conserved cell classes in neocortical neurogenesis and tracks developmental progression in progenitors and neurons. Examining outer radial glia markers across mammals, we find that transcriptomic elements of this key cell type likely arose in rodent gliogenic precursors before the full program emerged in the primate lineage.

Granular hydrogels offer a powerful platform for engineering porous, cell-instructive scaffolds with tunable mechanical, structural, and biochemical properties, yet introducing spatial and functional heterogeneity typically requires multiple microgel populations or complex fabrication strategies. Here, we present a programmable, photoresponsive granular hydrogel platform that enables post-assembly spatiotemporal control of scaffold mechanics and cell microenvironments from a single microgel formulation. Poly(ethylene glycol) microgels containing photolabile allyl sulfide moieties were synthesized via strain-promoted azide-alkyne cycloaddition and assembled into granular scaffolds capable of light-mediated remodeling through radical addition-fragmentation chain transfer. This chemistry afforded dynamic, on-demand, and spatially defined tuning of mechanical properties (G' = 0.7-3.7 kPa) while maintaining scaffold porosity (∼20%). High-resolution photopatterning across multiple length (6 µm-1 mm) and timescales enabled precise modulation of local microenvironments. Human mesenchymal stem/stromal cells embedded in these scaffolds responded to spatiotemporal modulation of matrix mechanics as observed by changes in morphology, yes-associated protein 1 (YAP) nuclear localization, and secretory profiles. Together, these results establish a versatile and broadly applicable strategy for programming mechanical heterogeneity and regulating cell behavior in granular hydrogels through photolabile moieties.

The Golgi complex serves as a critical hub for cellular homeostasis, yet its communication with the nucleus remains largely unexplored. By analyzing and siRNA-validating localization data from the Human Protein Atlas, we uncovered substantial proteome interconnectivity between the Golgi and nucleus, including an unexpected enrichment for DNA repair factors. We identify a cluster of DNA damage response (DDR) proteins occupying distinct sub-Golgi compartments that redistribute dynamically between the Golgi and nucleus in response to genotoxic stress, with the type of DNA lesion shaping the direction of redistribution. Focusing on the homologous recombination (HR) regulator RAD51C, we show that DNA damage triggers ataxia telangiectasia mutated (ATM)-dependent release of a giantin-tethered Golgi RAD51C pool, with subsequent importin-β-dependent nuclear import, where repair-associated foci form. Giantin depletion prematurely releases RAD51C, producing aberrant nuclear foci lacking key DDR markers, reducing ATM activation and HR efficiency, elevating genome instability, and accelerating proliferation. The Golgi thus acts as a spatiotemporal coordination node for DDR factors safeguarding genomic stability.

<h4>Introduction</h4>Translating cognitive test results into clinically meaningful measures of dementia severity remains challenging in real-world settings, where biomarkers are limited and functional assessment may occasionally be confounded by noncognitive factors. Although the Addenbrooke's Cognitive Examination-III (ACE-III) is widely used for detecting Alzheimer's disease (AD), its role in severity staging is less established. This study evaluated the usefulness of ACE-III for functionally anchored staging and compared it with the Mini-Mental State Examination (MMSE).<h4>Methods</h4>In a retrospective real-world cohort of 1,170 patients with AD dementia (late-onset, early onset, and mixed), severity was classified using the Functional Assessment Staging Tool (FAST). Receiver operating characteristic analyses were used to derive cut-offs for differentiating adjacent stages (FAST 4 vs. 5; FAST 5 vs. 6). Domain-level and subtype-specific analyses were conducted.<h4>Results</h4>ACE-III showed excellent discrimination between FAST 4 and 5 (area under the ROC curve [AUC] = 0.92; cut-off ≤56) and moderate discrimination between FAST 5 and 6 (AUC = 0.77; cut-off ≤42), outperforming MMSE in later stages (AUC = 0.63). Attention contributed most to earlier transitions, whereas later stages showed broader domain involvement. Cut-offs varied across AD subtypes.<h4>Conclusion</h4>ACE-III provides clinically meaningful estimates of dementia severity and may help overcome limitations of functional assessment in real-world settings, supporting its use for staging and monitoring in routine clinical practice.

<h4>Introduction</h4>Hepatocellular carcinoma (HCC) is a highly aggressive malignancy characterized by limited therapeutic options and significant cellular heterogeneity. This study aims to delineate the SUMOylation-mediated transcriptomic landscape and develop a robust prognostic signature to predict survival and immunotherapy response in HCC patients.<h4>Methods</h4>Malignant epithelial cells were classified into SUMOylation-related subtypes using nonnegative matrix factorization. A prognostic signature (SUMOylation.Sig) was constructed via machine learning and validated across multiple cohorts. Immune infiltration, immunotherapy response, and drug sensitivity were evaluated. Furthermore, we performed functional validation of a core signature gene, VPS72, through siRNA-mediated knockdown, colony formation and wound healing assays in HCC cells.<h4>Results</h4>We identified 10 prognostic SUMOylation-related genes (SRGs) and stratified malignant epithelial cells into five SUMOylation subtypes. The SUMOylation-C4 subtype was associated with favorable outcomes and enhanced immune infiltration. A 13-gene prognostic SUMOylation.Sig was developed and validated, effectively stratifying patients into low- and high-risk categories with distinct survival and responsiveness to immunotherapy. Low-risk patients exhibited increased immune cell infiltration and superior responses to immune checkpoint inhibitors. Experimental validation confirmed differential expression of signature genes in HCC cell lines. Notably, in vitro experiments demonstrated that VPS72 knockdown significantly suppressed the proliferation and migration of HepG2 and LM3 cells, confirming its oncogenic potential.<h4>Conclusions</h4>Our study establishes a SUMOylation-based cellular taxonomy and a robust 13-gene prognostic tool for HCC. The experimental validation of VPS72 underscores the functional importance of the signature, suggesting its potential involvement in tumor progression and highlighting potential therapeutic targets for personalized clinical management.

The 9p21 locus encodes critical cell cycle regulators p16^INK4A, p14^ARF, and p15^INK4B and contains a high density of SNPs associated with aging-related diseases. Progress toward understanding 9p21 regulatory mechanisms has been constrained by complex transcript structures and a shortage of cell models that retain physiologic locus control. We innovated tools to deconvolve the expression and regulation of 9p21 transcripts in single cells. We find that 9p21 transcripts exhibit highly divergent expression across tissues and cell types, with p15^INK4B predominating. Despite being frequently conflated under the term CDKN2A, p14^ARF and p16^INK4A are independently expressed across tissues, cell types, aging, and in a fibroblast model of replicative senescence. We annotate a network of cis-regulatory elements (CREs), whose activities and interactions underpin these complex regulatory dynamics. Our systematic characterization of 9p21 transcripts and CREs provides mechanistic insights and the tools for future studies of this vital locus.

Fibromyalgia is a common and debilitating chronic pain syndrome of poorly understood etiology. Here we conduct a multi-ancestry genome-wide association study meta-analysis across 2,563,755 individuals (54,629 cases and 2,509,126 controls) from 11 cohorts, identifying 26 risk loci for fibromyalgia. The strongest association was with a coding variant in HTT, the causal gene for Huntington's disease. Gene prioritization implicated the HTT regulator GPR52, as well as diverse genes with neural roles, including DCC, DRD2/NCAM1, MDGA2 and CELF4. Fibromyalgia heritability was exclusively enriched within brain tissues and neural cell types. Fibromyalgia showed strong, positive genetic correlation with a wide range of chronic pain, psychiatric and somatic disorders, including genetic correlations above 0.7 with low back pain, post-traumatic stress disorder and irritable bowel syndrome. Despite large sex differences in fibromyalgia prevalence, the genetic architecture of fibromyalgia was nearly identical between males and females. This study provides robust genetic evidence defining fibromyalgia as a central nervous system disorder, thereby establishing a biological framework for its complex pathophysiology and extensive clinical comorbidities.

Fertilization and early-cleavage failure are the major causes of assisted reproductive technology failure, but genotype-guided care is underused. We conducted whole-exome sequencing with Sanger confirmation in 20 men from couples with unexplained fertilization and cleavage failure after in vitro fertilization or intracytoplasmic sperm injection (ICSI). Then, further sperm transmission electron microscopy, immunofluorescence, and, where appropriate, embryo quantitative parental contamination test (qPCT) and kinship analysis were integrated in selected cases. Six novel variants were found in four families: two each in dynein axonemal heavy chain 10 ( DNAH10 ) and dynein axonemal heavy chain 12 ( DNAH12 ) and one in cilia and flagella associated protein 47 ( CFAP47 ) and phospholipase C zeta 1 ( PLCZ1 ), respectively. DNAH10/DNAH12 variants segregated with multiple morphological abnormalities of the sperm flagella and showed "9+2" axonemal disruption and reduced dynein signals. Hemizygous CFAP47 c.8006C>T is associated with cleavage failure and altered PLCZ1 localization. Homozygous PLCZ1 c.589C>T in a consanguineous pedigree coincided with multinuclear (≥3 pronuclei) zygotes, and qPCT/kinship indicated excess paternal contribution, consistent with suspected polyspermy in multinuclear zygotes. Management-matched genotype: ICSI restored fertilization in DNAH10/DNAH12 deficiency, whereas ICSI plus assisted oocyte activation (AOA) improved cleavage and yielded ongoing pregnancies in CFAP47/PLCZ1 deficiency. These findings outline potentially actionable clinical strategies-ICSI for axonemal dynein defects and ICSI plus AOA for activation pathway defects-and support targeted genetic testing plus embryo qPCT/kinship in selected cases.

<h4>Background</h4>Major depressive disorder (MDD) and amyotrophic lateral sclerosis (ALS) are usually treated as unrelated, yet depressive symptoms occur in a substantial minority of people with ALS and may appear early. These symptoms are heterogeneous and may reflect syndromal MDD, psychological and functional burden, fatigue, apathy, pseudobulbar affect, frontotemporal involvement, sleep or respiratory disturbance, medication effects, or shared affective vulnerability. A proposed pruning-continuum model suggests both disorders may share vulnerability in microglia-mediated synaptic pruning, with ALS amplified by autophagy and protein-quality-control failure and MDD by RNA-processing, stress, and immune dysregulation. We performed an exploratory secondary transcriptome-wide association study (TWAS)/pathway-integration analysis to test whether predefined nicotinamide mononucleotide (NMN)-nominated pathways map onto this vulnerability.<h4>Methods</h4>We integrated precomputed S-PrediXcan outputs for MDD and ALS across available brain-relevant tissues. Ten Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were predefined from a prior re-analysis of NMN-associated transcriptional programs in aged mouse metabolic tissues. Mouse-derived candidates were represented by human ortholog symbols before the human TWAS screen. The analysis tested nominated pathways rather than the 35-gene NMN-robust list as a standalone set. Cross-tissue screening used Stouffer Z aggregation, tissue-level Wilcoxon testing, competitive permutation testing, percentile bootstrap intervals, pairwise disease statistics, Levene variance tests, concordance measures, and leave-one-out sensitivity analysis. No analysis was treated as confirmatory or evidence of causal mediation.<h4>Results</h4>MDD showed the strongest Stouffer-based exploratory signal in the synaptic vesicle cycle pathway, with a meta-across-tissue Stouffer Z of 3.41 and a wide bootstrap 95% confidence interval of -0.46 to 7.40. This signal did not survive competitive permutation testing (p = 0.1222) or Wilcoxon testing (p = 0.1926). The strongest tissue-level result occurred in the amygdala (Z = 4.057; nominal Wilcoxon p = 0.0093), although tissue-level permutation testing was not performed in the multi-gene-set run. ALS showed no significant meta-across-tissue enrichment among the 10 nominated pathways but displayed candidate gene-level signals in autophagy, endosomal, and vesicle-related genes, including TBK1 and C9orf72. Exploratory Levene tests indicated variance heterogeneity in the regulation of the actin cytoskeleton, endocytosis, and neuroactive ligand-receptor interaction; the actin cytoskeleton and endocytosis remained significant in pooled global false discovery rate (FDR) analysis. Fourteen genes were influential in at least two focus pathways, including EGF, KNG1, FGF8, RAC1, PAK1, PAK2, RAF1, MAPK1, and FGFR1.<h4>Conclusions</h4>These findings are hypothesis-generating. MDD and ALS may stress overlapping cellular logistics processes while engaging largely different genes. MDD showed the strongest exploratory pathway-level signal in synaptic vesicle biology, whereas ALS showed candidate gene-level coherence in autophagy and endosomal processes without significant meta-pathway enrichment. NMN/NAD+ repletion is not established as a treatment for MDD, ALS, or their comorbidity. These findings generate hypotheses about NAD+-linked cellular stress pathways for future preclinical and clinical studies.

Large cell neuroendocrine carcinoma (LCNEC) is the rarest high-grade neuroendocrine carcinoma (NEC) of the urinary bladder. It can be observed in mixed neuroendocrine/non-neuroendocrine neoplasm (MiNEN), which is predominantly associated with urothelial carcinoma (UC) and less frequently with adenocarcinoma. Here, we report a primary MiNEN of the urinary bladder, composed of LCNEC and enteric-type adenocarcinoma. The patient was a 58-year-old male who initially presented with painless hematuria and dysuria. Transabdominal ultrasonography demonstrated a 4.8 × 2.4 cm hypoechoic lesion arising from the posterior bladder wall. Histologically, the tumor showed distinct yet closely apposed components of LCNEC and enteric-type adenocarcinoma. Immunohistochemically, the LCNEC component was positive for neuroendocrine markers, including CD56, chromogranin, synaptophysin, and INSM1, while the adenocarcinoma component was positive for CK20 and CDX2. Notably, both components showed aberrant P53 expression (P53-null pattern), supporting a shared clonal origin with divergent differentiation. After progression following initial platinum-based chemotherapy, the patient achieved a radiographic partial response (PR) of pulmonary metastases after treatment with enfortumab vedotin plus pembrolizumab administered with radiotherapy. Our case highlights the diagnostic importance of recognizing a high-grade neuroendocrine component in bladder tumors. Furthermore, antibody-drug conjugates and immune checkpoint inhibitors may merit further study in this rare entity.

Although chimeric antigen receptor T (CAR-T) cell therapy has achieved remarkable success in hematological malignancies, its therapeutic efficacy in solid tumors remains limited by several challenges, including insufficient tumor infiltration, T cell exhaustion and the immunosuppressive tumor microenvironment (TME). CRISPR/Cas, a third-generation gene editing technology developed in recent years, is characterized by its simplicity and high efficiency. This technology has demonstrated broad application potential across multiple fields and has emerged as a powerful tool for improving CAR-T cell therapy. In this review, we summarize recent advances in the application of CRISPR/Cas gene editing technology to enhance the antitumor activity of CAR-T cells against solid tumors. We also discuss the key challenges currently faced and systematically propose potential strategies for overcoming the limitations.

Spinal muscular atrophy (SMA) is a devastating autosomal recessive neuromuscular disorder characterized by progressive degeneration of lower motor neurons, leading to muscle weakness and atrophy predominantly manifesting in childhood. The disease is caused by homozygous deletion or loss-of-function mutations in the survival motor neuron 1 (SMN1) gene, with the nearly identical SMN2 gene acting as a critical modifier of disease severity. Since the initial clinical descriptions in 1891 and the identification of the causative gene in 1995, the field has witnessed transformative therapeutic progress. The FDA approval of nusinersen, an antisense oligonucleotide targeting SMN2 splicing, in 2016 marked the first disease-modifying therapy, followed by the gene replacement therapy onasemnogene abeparvovec in 2019 and the orally administered small-molecule splicing modifier risdiplam in 2020. These SMN-targeted therapies have substantially improved survival and motor outcomes, particularly when administered presymptomatically as enabled by increasingly widespread newborn screening programs. However, a significant proportion of patients, especially those with later-onset disease and adults, show limited responses, and existing therapies cannot reverse established neurodegeneration or muscle pathology. In this review, we trace the history of SMA research and therapy development, critically compare the three approved treatments, and discuss key unresolved challenges including treatment-switching criteria, management of late-diagnosed patients, and the emerging role of biomarkers such as neurofilament light chain and compound muscle action potential. We further explore non-SMN therapeutic strategies targeting PTEN, plastin 3, neurocalcin delta, myostatin, and neuroinflammation, and examine the rationale for combination approaches that integrate SMN restoration with muscle- and neuro-protective agents. Finally, we address future directions encompassing precision medicine, next-generation gene editing, and biomarker-driven trial design. While SMA has been transformed from a fatal childhood disorder to a treatable condition, a definitive cure for all patients remains the goal of ongoing and future research.

<h4>Background</h4>Hypopituitarism is associated with adverse body composition and steatotic liver disease, particularly when the growth hormone/insulin-like growth factor-1 (GH/IGF-1) axis is disturbed. Progression to decompensated cirrhosis is uncommon, and published reports do not establish a direct causal relationship.<h4>Case presentation</h4>A 37-year-old woman presented with jaundice, massive ascites, splenomegaly, thrombocytopenia, hypoalbuminemia, coagulopathy, and hyponatremia. She had undergone pituitary adenoma surgery at approximately 12 years of age, followed by amenorrhea and more than two decades without regular endocrine follow-up or hormone replacement. The clinical and biochemical profile supported long-standing panhypopituitarism. Possible central HPA-axis dysfunction was present; secondary adrenal insufficiency was clinically probable but not dynamically confirmed. Other features included central hypothyroidism, hypogonadotropic hypogonadism, low prolactin, central diabetes insipidus, and severe GH/IGF-1 axis disturbance. GH stimulation testing was not performed, and interpretation of the GH/IGF-1 axis was confounded by advanced liver disease. Common viral, autoimmune, alcohol-related, drug-related, and overt cardiac causes were not supported. However, transferrin saturation was 94.6%, and iron overload, Wilson disease, alpha-1 antitrypsin deficiency, and histological metabolic steatohepatitis were not fully excluded. Liver biopsy supported advanced fibrosis/cirrhosis, and explant pathology showed micronodular cirrhosis with hepatocellular cholestasis. She underwent liver transplantation in May 2026. Aminotransferase levels subsequently declined, but persistent thrombocytopenia, pulmonary infection, and acute kidney injury requiring hemodialysis developed. She progressed to septic shock and died more than 20 days after transplantation.<h4>Conclusion</h4>Long-standing untreated panhypopituitarism may have contributed to metabolic and fibrogenic risk in this patient, but incomplete etiological evaluation precluded a definitive causal link. Lifelong endocrine follow-up and attention to liver health are warranted after pituitary surgery.

<h4>Background</h4>Glioma is a highly aggressive brain tumor with poor prognosis, necessitating new biomarkers and therapeutic targets. This study aimed to explore the role of Pit-Oct-Unc class 3 homeobox 4 (POU3F4) in glioma using bioinformatics analysis and experimental verification.<h4>Methods</h4>Bioinformatics analysis was performed using the public databases The Cancer Genome Atlas (TCGA) and Human Protein Atlas database to explore POU3F4 expression and prognosis in gliomas. Samples were collected from clinical glioma tissues, paracancerous tissues, and plasma of patients and healthy candidates. POU3F4 expression was examined using molecular biological methods, and its potential diagnostic value was assessed. <i>In vitro</i>, normal human astrocytes (NHAs) and various glioma cells (U87MG, U251MG, T98G, A172, and LN229) were used to assess POU3F4 expression using quantitative polymerase chain reaction (qPCR) and western blotting (WB). U87MG and U251MG cells were transfected with specific small interfering RNAs (siRNAs) of POU3F4. The effects of POU3F4 siRNA on cell proliferation, apoptosis, and migration were assessed using the Cell Counting Kit-8 (CCK-8), clone formation, 5-ethynyl-2'-deoxyuridine (EdU), terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL), wound healing, and Transwell assays. The binding of KLF15 to the promoter region of POU3F4 was validated using chromatin immunoprecipitation (ChIP) assay. Finally, the molecular mechanism of KLF15-POU3F4 axis in regulating the PI3K/AKT signaling pathway was preliminarily explored by measuring the phosphorylation levels of phosphatidylinositol 3-kinase (PI3K) and protein kinase B (AKT).<h4>Results</h4>Bioinformatics analysis revealed that POU3F4 levels were increased in gliomas, and upregulated POU3F4 may be associated with poor prognosis. Additionally, clinical sample analysis confirmed that POU3F4 was overexpressed in glioma tissues and plasma samples, and its levels in glioma tissues were positively associated with those in the plasma. The receiver operating characteristic (ROC) curve demonstrated that POU3F4 levels in both tissue and plasma exhibited good diagnostic value for glioma. <i>In vitro</i> experiments demonstrated that POU3F4 was overexpressed in different gliomas. POU3F4 knockdown reduced the growth and metastasis of glioma cells and increased apoptosis. Mechanistic studies demonstrated that POU3F4 knockdown did not affect the messenger ribonucleic acid (mRNA) or protein expression of PI3K or AKT but significantly reduced their phosphorylation. In addition, JASPAR database prediction and ChIP experiments confirmed that KLF15 is an upstream transcription factor of POU3F4, which can directly bind to the POU3F4 promoter region and positively regulate its expression. Functional recovery experiments have shown that the KLF15-POU3F4 axis promotes the proliferation, migration, and invasion of glioma cells by activating the PI3K/AKT signaling pathway.<h4>Conclusions</h4>POU3F4 was upregulated in glioma and facilitated glioma cell growth and metastasis while suppressing apoptosis. KLF15 is its upstream transcription factor that positively regulates the expression of POU3F4. The KLF15-POU3F4 axis promotes glioma progression by activating the PI3K/AKT signaling pathway.

Parkinson's disease (PD) is defined pathologically by loss of dopamine-producing neurons in the substantia nigra pars compacta (SNc). Yet synaptic dysfunction emerges much earlier, making it essential to define the mechanisms that drive early nigrostriatal deregulation. In the SNc, molecularly distinct dopamine neuron subtypes show differential susceptibility to PD. Here, we used intersectional genetic mouse models to determine how the PD-linked kinase LRRK2 affects vulnerable dopamine subtypes. Immunofluorescence and proximity-labeling proteomics revealed enriched LRRK2 expression in vulnerable dopamine neuron subclusters. High-resolution imaging showed that pathogenic LRRK2 disrupts presynaptic release-site organization in vulnerable dopamine axons, leading to reduced spontaneous and evoked striatal dopamine release in vivo. Proteomic analyses further showed that mutant LRRK2 increases phosphorylation of RAB3 proteins, impairing their interaction with the active-zone effectors RIM1 and RIM2. Together, these findings highlight a subtype-specific, cell-autonomous mechanism by which pathogenic LRRK2 impairs PD-vulnerable nigrostriatal synapses and provide a framework for therapeutic strategies targeting early synaptic deficits in PD.

<h4>Background</h4>Coronary artery disease (CAD) is a leading cause of death worldwide. Evidence of genetic predisposition is derived mostly from studies of Europeans and East Asians.<h4>Objectives</h4>To test selected variants for association with early-onset CAD.<h4>Methods</h4>We performed a case-control study of 697 young angiography-defined CAD cases (women aged <55 years, men <45) and 911 age- and sex-matched controls in Iran. We tested 24 variants (12 established CAD loci, 12 candidates in hemostasis genes) using age- and sex-adjusted logistic regression, stratified by ethnicity. A 6-variant unweighted genetic risk score (GRS) was built from the top loci. Additive interaction with cardiovascular risk factors was assessed by relative excess risk due to interaction.<h4>Results</h4>Of 12 established loci, <i>HCG27</i>-<i>HLA-C</i>, <i>CDKN2A</i>/<i>CDKN2B</i>, and <i>SORT1</i> showed nominal or near-nominal replication and 4 were directionally aligned with prior literature, whereas 5 showed no association. Among hemostasis candidates, <i>F5</i> rs4524 was nominally associated with increased risk, and <i>TSPAN15</i> rs78707713 and <i>F11</i> rs2289252 suggested reduced risk. Ethnicity-stratified analyses indicated nominal or suggestive ancestry-specific effects at <i>SORT1</i>, <i>HCG27</i>-<i>HLA-C</i>, <i>CDKN2A</i>/<i>CDKN2B</i>, <i>LPA</i>, <i>VWF</i>, <i>MIA3</i>, and <i>CXCL12</i>. The GRS (range, 2-11 alleles/person) yielded a per-allele odds ratio of 1.20 (95% CI, 1.12-1.29), ie, a 5.2-fold higher risk across the observed range. Combining GRS with metabolic comorbidities showed super-additive effects (relative excess risk due to interaction, 6.66; 95% CI, 2.04-11.28).<h4>Conclusion</h4>Ten variants, including 3 previously linked to venous thrombosis, showed nominal or suggestive association with early-onset CAD in Iranians or specific ethnic groups. Genetic burden substantially amplified risk in the presence of metabolic comorbidities. Our findings underscore the need for ancestry-aware studies in multiethnic populations.

TP53 -mutated acute myeloid leukemia (AML) has dismal outcomes with current treatments and represents a critical unmet need. TP53 -mutated AML is proposed to be susceptible to immunotherapeutic approaches but, to date, there is no established immunotherapy for this sub-group. Expression of stimulator of interferon genes (STING), a key innate immune driver that activates interferon (IFN) signaling, is decreased by epigenetic silencing or mutation in many cancers, including those with TP53 mutations. Here, we report that response to the next-generation synthetic STING agonist C92 is potentiated in AML cell lines and primary cells with TP53- mutated versus wild-type (WT) cells, representing a previously undescribed vulnerability of these leukemia cells to STING small molecule therapies. Moreover, combining treatment with the DNA methyltransferase inhibitor (DNMTi) decitabine (DAC), significantly increases STING activation, with marked transcriptome-wide increase in repetitive elements (REs) and upregulation of a critical set of interferon-related genes. Cell death in TP53 KO versus WT AML is specifically dependent on innate immune zinc finger NFX1-type containing 1 (ZNFX1) and Z-DNA-binding protein 1 (ZBP1) driving increased cleavage and activation of Receptor-Interacting-Serine/Threonine-Protein Kinase 3 (RIPK3) and mixed lineage kinase domain-like protein (MLKL), suggesting mechanisms of necroptosis. Finally, C92 and DAC combination significantly reduces leukemia burden in humanized AML mouse models, accompanied by increased immune responses, including cytokines and cytotoxic T lymphocytes in the leukemia microenvironment. These results support development of clinical trial strategies combining STING agonists with DNMTis for patients with TP53 -mutated AML. <h4>Summary</h4> TP53-mutated AML potentiates effects of novel next-generation STING agonist C92, with unique allosteric and non-cyclic dinucleotide (non-CD) mechanism of action, inducing increased STING activation and cytokine release STING agonists and DNMTis, synergistically increase STING activation with marked transcriptome-wide increase in repetitive elements (REs) and upregulation of a critical set of interferon-related genes in TP53 -mutated AML STING agonists induce necroptosis via a STING-ZNFX1-ZBP1-necroptosis axis in TP53 -mutated AML. This drug combination reduces leukemia burden, activates immune responses in AML models and supports translation for high-risk AML patients. <h4>Statement of Translational Relevance</h4> This pre-clinical study identifies a novel therapeutic vulnerability in ( TP53 )-mutated acute myeloid leukemia (AML), a poor prognosis subtype with a critical unmet need. Novel next-generation STING agonist C92, with unique allosteric and non-cyclic dinucleotide (non-CD) mechanism of action, induces increased STING activation and cytokine release, compared with WT TP53 in AML cell lines and primary cells, and has superior STING activity with respect to several STING agonists currently in clinical studies. Combining C92 treatment with the DNA methyltransferase inhibitor (DNMTi) decitabine (DAC) synergistically increases STING activation, with marked transcriptome-wide increase in repetitive elements (REs) and upregulation of a critical set of interferon-related genes, driving ZNFX1-driven inflammatory necroptotic cell death. Utilizing humanized mouse models, C92 in combination with DAC significantly reduces leukemia burden and enhances cytotoxic T-cell responses in the tumor microenvironment, supporting clinical translation for high-risk AML patients.

Traumatic brain injury (TBI) is a prevalent cause of secondary neuropsychiatric disorders, with post-traumatic anxiety significantly worsening long-term outcomes. However, the key neural circuits and molecular mechanisms underlying post-TBI anxiety remain poorly understood. Here, we show that TBI patients with frontal lobe injuries and anxiety exhibited increased activity and enhanced functional connectivity in the anterior insular cortex (aIC) and lateral orbitofrontal cortex (lOFC). Mechanistically, we established a mouse model of moderate frontal lobe TBI, categorizing animals into anxious and non-anxious TBI groups. BOLD-fMRI and c-Fos expression analyses revealed that mice with post-traumatic anxiety also exhibited increased activity in the aIC and lOFC, as well as heightened activity in the aIC-lOFC glutamatergic circuit. Optogenetic and chemogenetic modulation of this circuit bidirectionally regulated post-traumatic anxiety. Single-cell RNA sequencing of aIC glutamatergic neurons identified a specific downregulation of KCNC3 in anxious TBI mice. Functional validation confirmed that KCNC3 downregulation mediates hyperactivity in the aIC-lOFC glutamatergic circuit and induces post-traumatic anxiety. Together, these findings unveil a critical role of TBI‑induced KCNC3 downregulation in aIC glutamatergic neurons in driving hyperactivity of the aIC-lOFC glutamatergic circuit and resulting post‑traumatic anxiety, and they suggest KCNC3 and the aIC-lOFC circuit as promising therapeutic targets for post‑TBI anxiety.

Myelination, driven by differentiation of oligodendrocyte precursor cells, is critical for metabolic and structural support and efficient axonal signal transmission in neurons. Loss of myelin is a hallmark of multiple sclerosis and other devastating demyelinating disorders. As demyelination persists, neurons become increasingly vulnerable, leading to neurodegeneration and chronic disability. Restoring myelin through endogenous repair mechanisms offers a promising therapeutic approach to mitigate progressive neuronal loss. One key regulator of myelination is the G protein-coupled receptor 17, GPR17, whose chronic upregulation in oligodendrocyte precursor cells is commonly seen with myelin injury. In line with single-nucleus transcriptomic data showing predominant expression of GPR17 in committed oligodendrocyte precursor cells, our postmortem immunohistochemical analyses of MS patient tissue revealed a significant upregulation of GPR17+/BCAS1+ oligodendrocyte precursor cells adjacent to and in demyelinated lesions. Importantly, remyelinated lesions lacked GPR17 immunoreactivity, consistent with a model in which sustained GPR17 expression is associated with demyelination and impaired oligodendrocyte precursor cell differentiation. To test the impact of pharmacological GPR17 inhibition on remyelination, we evaluated the effects of a novel, selective GPR17 antagonist in cuprizone-induced murine demyelination models. This toxin-induced approach has been widely used to study mechanisms of de- and remyelination, in the absence of the full inflammatory complexity of demyelinating diseases such as multiple sclerosis. We show that oral treatment results in robust functional recovery consistent with remyelination, as evidenced by improved spatial memory and recovery of visual evoked potential latency delays. GPR17 antagonism also accelerated structural remyelination in the corpus callosum and optic nerve. Together, these findings support a role for pharmacological GPR17 antagonism in promoting remyelination and highlight this G protein-coupled receptor as a promising therapeutic target for demyelinating disorders.

Kaposi sarcoma-associated herpesvirus (KSHV) is an oncogenic gammaherpesvirus that is associated with several human malignancies, and primary infection by KSHV can activate both DNA and RNA sensing pathways. Host innate immunity is a rapid, first-line defense against foreign pathogens, where pattern recognition receptors detect virus-associated antigens and activate the type I interferon response to prime the cell to an antiviral state to limit viral infections. However, viruses such as gammaherpesviruses, have evolved mechanisms to co-opt host factors to antagonize these pathways to promote persistent infection. The cellular vaccinia-related kinase (VRK) family is a trio of serine/threonine kinases, VRK1, VRK2 and VRK3, named for their homology to the vaccinia virus B1 kinase. The VRKs are involved in many cellular processes, including nuclear envelope assembly during mitosis and the DNA damage response. However, the role of the VRKs in KSHV infection and innate immunity has not previously been explored. We found that VRK3 is upregulated in response to KSHV primary infection of endothelial cells. VRK3 is also upregulated during KSHV reactivation from latency. Knockdown of VRK3 limited KSHV primary infection and lytic reactivation and resulted in increased activation of TBK1 and IRF3 and subsequent upregulation of interferon stimulated genes. Overexpression of VRK3 reversed this phenotype. Furthermore, we are also the first to demonstrate that VRK3 is a negative regulator of the interferon response even in the absence of viral infection. Collectively, our data highlight the importance of the host factor, VRK3, as a novel suppressor of the type I interferon response during the KSHV viral lifecycle.

Bronchopulmonary dysplasia (BPD), a lung disease associated with preterm birth, is characterized by arrested alveolarization and impaired pulmonary vascular growth. Exposure to early life hyperoxia is a disease-contributing factor, and endothelial determinants of injury versus repair remain incompletely defined. We hypothesized that endothelial-specific loss of <i>Hif-1α</i> during the saccular stage of development would adversely impact lung development and augment injury after hyperoxia exposure. Tamoxifen-inducible endothelial-specific <i>Hif-1α</i> knockout and <i>Hif-1α</i>^fl/fl littermates were exposed to room air or 95% O₂ from <i>postnatal day</i> (PND) <i>1</i>-<i>5</i>, and lungs were analyzed at PND7 and PND21 for morphometry, vascular density, and lung endothelial transcriptomic response. Endothelial <i>Hif-1α</i> deletion had no impact on baseline alveolarization but reduced vascular density under normoxia and significantly worsened hyperoxia-induced alveolar simplification, septal thickening, and vascular rarefaction. Hyperoxia increased endothelial proliferation in both genotypes after 48 h of recovery in normoxia. Bulk RNA sequencing of the lung endothelial cells revealed a shared hyperoxia injury program (e.g., <i>Inhba</i>, <i>Serpine1</i> induction; <i>Esm1</i>, <i>Gpihbp1</i>, <i>Car2</i> suppression). The transcriptomic response in <i>HIF1α</i>^ECKO endothelial cells was significantly muted compared with wild-type endothelial cells, lacking induction of <i>Hmox1</i>, <i>Prdx6</i>, and <i>Anxa1</i>. <i>Hif-1α</i>-deficient endothelial cells upregulated <i>Col18a1</i> and <i>Ackr3</i> and showed enrichment of TNF-NF-κB, TGF-β, and cell-cycle checkpoint pathways, consistent with maladaptive remodeling. Cross-referencing with human single-cell BPD data demonstrated that genes identified in the murine hyperoxia model exhibit conserved but heterogeneous endothelial expression patterns across BPD disease states. These findings identify endothelial <i>Hif-1α</i> as a central determinant of recovery after neonatal hyperoxia exposure.<b>NEW & NOTEWORTHY</b> This study identifies endothelial hypoxia-inducible factor-1α (HIF-1α) as a critical determinant of neonatal lung response to hyperoxia. Using an inducible endothelial-specific <i>Hif-1α</i> knockout model, we demonstrate that loss of endothelial <i>Hif-1α</i> exacerbates hyperoxia-induced impairments in pulmonary vascular development and alveolarization, key pathological features of bronchopulmonary dysplasia.

<h4>Background</h4>Intestinal ageing is characterized by progressive epithelial deterioration, barrier dysfunction and impaired regenerative capacity, which together contribute to age-related intestinal decline. Sulfated fucans derived from Saccharina japonica (SJ-FUC) are marine polysaccharides with potential bioactivities, although their role in intestinal ageing remains unclear. This study investigated the protective effects of SJ-FUC in a D-galactose-induced ageing mouse model.<h4>Results</h4>Administration of SJ-FUC significantly alleviated intestinal ageing-associated phenotypes by reducing oxidative stress, suppressing senescence-associated secretory phenotype-related inflammatory cytokines, and downregulating the senescence markers P53, P16 and P21. Histological analysis showed marked improvement in ileal architecture and epithelial barrier integrity. SJ-FUC also mitigated ageing-associated regenerative decline, as indicated by increased expression of intestinal stem cell-related markers Lgr5, Ascl2 and Olfm4, elevated numbers of Olfm4-positive and Ki67-positive cells in ileal crypts, and restoration of Wnt/β-catenin signalling. In addition, SJ-FUC promoted goblet and Paneth cell differentiation. 16S rRNA sequencing further showed that SJ-FUC remodelled gut microbial composition, characterized by enrichment of beneficial taxa such as Lactobacillus and reduction of potentially detrimental bacteria including Bacteroides.<h4>Conclusion</h4>Sulfated fucans from Saccharina japonica alleviate intestinal ageing by preserving epithelial integrity and restoring regenerative capacity, supporting their potential as functional marine polysaccharides for improving age-related intestinal dysfunction. © 2026 Society of Chemical Industry.

No abstract available.

<h4>Objectives</h4>To evaluate whether the spatial pattern of obstructive sleep apnea (OSA)-related regional magnetic susceptibility alterations colocalizes with the normative distribution of neurotransmitter receptors and transporters.<h4>Methods</h4>Regional magnetic susceptibility was assessed in 36 OSA patients and 30 screen-negative controls (SNC) using quantitative susceptibility mapping (QSM). The spatial colocalization between OSA-related QSM alterations and the normative distribution of receptors/transporters was examined using PET-derived neurotransmitter maps included in NiSpace. Clinical correlates of colocalization strength were also explored.<h4>Results</h4>Compared with SNC, patients with OSA showed increased magnetic susceptibility in the left insula, left middle temporal gyrus, and left inferior temporal gyrus (all P_FDR = 0.047), with similar trends in the right putamen and right transverse temporal gyrus that did not survive correction. These QSM alterations colocalized with the normative distribution of NMDA receptor (mean Rho = 0.177, P_Meff = 0.001), Dopamine transporter (DAT) (mean Rho = 0.149, P_Meff = 0.040), and 5-HTT (mean Rho = 0.153, P_Meff = 0.040). The strength of colocalization with NMDA receptors, DAT, and 5-HTT was significantly associated with disease severity.<h4>Conclusion</h4>This study provides convergent evidence linking OSA-related magnetic susceptibility alterations, consistent with iron-related changes, to the normative distribution of glutamatergic, serotonergic, and dopaminergic neurotransmitter systems. These spatial associations correlated with disease severity, highlighting the potential of spatial colocalization approaches for understanding neurochemical vulnerability in OSA.

<h4>Background</h4>Deer antler is the only mammalian organ capable of periodic complete regeneration. Its astonishing growth rate-reaching several centimeters per day-provides a unique model for research in tissue regeneration and developmental biology. This rapid growth relies on the protective and signaling functions of the skin, as well as the vigorous proliferation and differentiation capacity of mesenchymal cells. Although previous studies have identified some key factors involved in antler growth, systematically comparing the transcriptomic profiles of these two core tissues (skin and mesenchyme) to identify candidate genes regulating their synergistic growth is crucial for deciphering the molecular mechanisms underlying this "ultra-fast" growth.<h4>Methods</h4>The Illumina NovaSeq 6000 sequencing platform was used to perform transcriptome sequencing on the antler tissues of the <i>Cervus elaphus xanthopygus</i> (Dongbei red deer) with a growth period of approximately 60 days. Bioinformatics methods were used to identify the transcriptional expression of genes in the antler tissues, and the expression levels of some genes were tested by fluorescence quantitative PCR.<h4>Results</h4>A total of 109.82 Gb of transcriptome data was generated from the skin and mesenchymal tissues of antlers, and 40,023 transcripts were obtained. There were 3,530 genes with differential expression between the skin and the mesenchymal tissues of antlers. Genes with upregulated expression levels in the mesenchymal layer were enriched in phosphate-containing compound metabolic processes and multiple biological processes related to bone development such as skeletal system development, cartilage development, regulation of ossification, and enamel mineralization. Among the genes enriched in the cancer pathway and the PI3K-Akt signaling pathway, those that may play an important role in the rapid growth process of deer antlers were screened, including <i>MDM2</i>, <i>RPS6KB1</i>, <i>CASP3</i>, <i>BMP4</i>, <i>PTCH1</i>, <i>SOX-6</i>, <i>YWHAZ</i>, and <i>HSP90B1</i>.<h4>Conclusions</h4>The discovery of these genes provides a reference for a further in-depth understanding of the mechanism of the rapid growth of antlers.

<h4>Background</h4>The antler primary growth center, located at the distal tip of the growing antler, comprises five consecutive tissue zones beneath the velvet skin. Because the outermost reserve mesenchyme contains blastema progenitor cells with multipotent differentiation capacity, antler regeneration recapitulates embryonic skeletal development through endochondral ossification (ECO). The molecular mechanisms governing cell fate decisions and tissue morphogenesis across these zones remain poorly understood.<h4>Methods</h4>We profiled the proteome of the sika deer (<i>Cervus nippon</i>) antler growth center across all five tissue zones using the Orbitrap Astral platform in data-independent acquisition (DIA) mode. Protein identification and quantification were performed with DIA-NN. Proteome dynamics were analyzed by integrating three complementary clustering approaches including weighted gene co-expression network analysis (WGCNA), Fuzzy C-means clustering, and monotonic feature selection. Differentially expressed proteins were annotated by gene ontology (GO) enrichment analysis. Transcriptomic data from the same tissue zones were reanalyzed to assess mRNA-protein concordance.<h4>Results</h4>A total of 8,173 proteins were identified across the five tissue zones, representing the most comprehensive proteomic coverage of the antler growth center. Clustering analyses resolved distinct protein expression modules corresponding to sequential ECO stages from mesenchymal proliferation in the reserve mesenchyme to cartilage mineralization in the innermost zone. Deep proteome coverage enabled detection of low-abundance chondrogenic transcription factors SOX9, SOX6, and RUNX3, whose spatial expression matched their known roles in chondrogenesis. At the protein level, the reserve mesenchyme co-expressed 11 embryonic and 23 mesenchymal stem cell markers, indicating that antler stem cells represented a unique mesenchymal stem cell (MSC) population possessing partial embryonic stem cell (ESC)-like features.<h4>Conclusions</h4>This study provides a spatially resolved proteomic atlas of the complete antler growth center and identifies key regulatory proteins at each stage of endochondral ossification. The findings offer new insights into progenitor cell characteristics, transcription factor activity, and protein dynamics during bone development, with potential relevance to bone repair and regenerative medicine.

Fe(0) dinitrogen complexes [SiP₃ ^R]Fe-N[triple bond, length as m-dash]N-Fe[SiP₃ ^R] (1-R; [SiP₃ ^R] = PhSi(CH₂PR₂)₃; R = ^<i>i</i> Pr, Ph) are accessed by one- or two-electron reduction of Fe(i) and Fe(ii) chloride complexes [SiP₃ ^{^<i>i</i> Pr}]FeCl or κ²-[SiP₃ ^R]FeCl₂ (R = ^<i>i</i> Pr, Ph), respectively, and fully characterized by means of multinuclear NMR spectroscopy, single-crystal X-ray diffraction (SC-XRD), and Mössbauer spectroscopy. Complexes 1-R represent rare examples of bimetallic Fe(0) compounds with dinitrogen as a bridging ligand and have an overall spin of <i>S</i> = 0. Heating 1-R in the presence of SiH₄ gas resulted in four-fold Si-H activation to furnish diiron complexes [SiP₃ ^R](H)₂Fe[double bond, length as m-dash]Si[double bond, length as m-dash]Fe(H)₂[SiP₃ ^R] (2-R; R = ^<i>i</i> Pr, Ph), which were characterized by multinuclear NMR spectroscopy, SC-XRD, Mössbauer spectroscopy, and density functional theory calculations. Complexes 2-R exhibit the shortest iron-silicon bonds to date and nearly linear Fe[double bond, length as m-dash]Si[double bond, length as m-dash]Fe cores. This work extends a previously developed strategy for SiH₄ activation from cobalt to iron, while offering insight into the silicon-atom extrusion from SiH₄ in molecular systems.

Using single-cell transcriptomics of bronchoalveolar lavage cells from <i>Mtb</i>/SIV co-infected rhesus macaques on cART, we reveal profound immune dysregulation during early SIV co-infection of latent tuberculosis. SIV induces a sharp decline in CD4⁺ T cells, NK, and NKT cells, with incomplete recovery of <i>Mtb</i>-specific T_H1 effector responses despite viral suppression. Instead, a persistent T_H17-skewed environment emerges, alongside sustained myeloid inflammation driven by Type I interferon signaling and pro-inflammatory regulators such as <i>KLF6</i> and <i>NFKB1</i>. Ligand-receptor network analyses demonstrate expanded CD4⁺ T cell-macrophage crosstalk and loss of immune homeostasis that cART fails to fully restore. These findings expose how SIV remodels the pulmonary immune landscape to impair protective immunity against <i>Mtb</i>, providing a transcriptomic framework to explain TB reactivation in HIV infection. Our work highlights the urgent need for adjunctive immunotherapies to complement cART, aiming to rebalance immune responses and improve TB control in co-infected individuals.

Males are more commonly diagnosed with autism spectrum disorder (ASD) than females with a ratio of about 4-1. However, the neural mechanisms underlying the sex differences in ASD are unknown. Social deficits are the core symptoms of patients with ASD. Previous studies showed that diminished activity-dependent brain-derived neurotrophic factor (BDNF) signaling induced differential severity of autism-like social preference deficits in male and female mice by using a mouse model with genetic knock-in of human BDNF methionine (Met) allele, which significantly decreased activity-dependent BDNF release without affecting basal BDNF secretion. Here, we investigated the synaptic mechanisms for diminished activity-dependent BDNF-induced differential severity of social preference deficits in males and females. The prefrontal cortex (PFC) is a critical brain region for social behaviors. Whole-cell patch-clamp brain slice recordings showed that diminished activity-dependent BDNF signaling differentially increased the frequency of spontaneous action potentials (sAPs) of pyramidal neurons in the PFC of male and female BDNF^+/Met mice. The frequency of sAPs in male BDNF^+/Met mice was higher than in female BDNF^+/Met mice. Diminished activity-dependent BDNF signaling differentially enhanced excitatory synaptic transmission and dampened inhibitory synaptic transmission of pyramidal neurons at pre- and post- synapses in males and females, which were mediated by dysregulated transcriptional levels of key synaptic genes. Chemogenetic inhibition of pyramidal neurons in the PFC of BDNF^+/Met mice was sufficient to ameliorate autism-like social preference deficits in males and females. This study reveals synaptic mechanisms underlying the differential severity of social preference deficit in male and female BDNF^+/Met mice, which provides a potential neural basis for sex differences in male and female ASD patients with and without the BDNF Val66Met SNP.

<b>Purpose:</b> To characterize the messenger RNA (mRNA) and microRNA (miRNA) expression profiles of the normal human cornea and conjunctiva using impression cytology (IC) samples and to investigate their molecular characteristics and regulatory networks. <b>Methods:</b> Corneal and conjunctival IC samples were collected from healthy subjects. Whole-transcriptome and miRNA sequencing were performed, followed by differential expression and bioinformatics analyses. Regulatory networks, protein interaction networks, and functional enrichment analyses were constructed. Selected genes and miRNAs were validated by RT-qPCR. <b>Results:</b> A total of 1676 differentially expressed genes and 175 differentially expressed miRNAs were identified between the cornea and conjunctiva. Functional analyses revealed that genes showing higher expression in the cornea were mainly associated with epithelial structure, barrier function, and antiviral immune responses. In contrast, genes showing higher expression in the conjunctiva were primarily involved in immune regulation, secretion, metabolic detoxification, and tissue remodeling. PPI network analysis showed that hub genes in the cornea were predominantly interferon-stimulated genes related to antiviral responses, while those in the conjunctiva were mainly involved in cell cycle regulation and metabolic detoxification. GO and KEGG analyses further supported these functional distinctions. RT-qPCR validation generally supported the expression patterns identified by RNA sequencing. <b>Conclusions:</b> Our findings reveal that the cornea is characterized by gene expression programs supporting epithelial homeostasis, barrier function, and antiviral immunity, whereas the conjunctiva exhibits transcriptional signatures related to immune surveillance, secretion, metabolic processing, and tissue remodeling. The miRNA-mRNA regulatory networks constructed in this study provide new insights into the molecular regulatory mechanisms of the ocular surface and offer a theoretical basis for future research into disease mechanisms and targeted therapeutic strategies.

<h4>Background</h4>Nasopharyngeal carcinoma (NPC) is a malignant tumor of the nasopharyngeal mucosal epithelium. The aryl hydrocarbon receptor (AhR)-a widely distributed nuclear receptor regulating adaptive adverse responses-is confirmed carcinogenic upon activation. Thus, this study aims to identify novel biomarkers for NPC treatment.<h4>Methods</h4>Relevant data were obtained from public databases. Biomarkers were developed through the application of differential expression analysis, univariate Cox regression, and machine learning algorithms. Molecular mechanisms were then explored through immune infiltration and molecular regulatory networks. Additionally, the drug sensitivity analysis revealed potential differences in sensitivity to chemotherapy drugs. The experimental verification was conducted using reverse transcription quantitative polymerase chain reaction (RT-qPCR).<h4>Results</h4>Cholinergic receptor nicotinic beta 2 subunit (CHRNB2), adenylate cyclase 4 (ADCY4), and cell division cycle 6 (CDC6) were identified as biomarkers. CHRNB2 and CDC6 were highly expressed, while ADCY4 was lowly expressed in NPC. ADCY4 showed a significant positive correlation with plasmacytoid dendritic cells [correlation coefficient (cor) =0.48, P<0.001]. In contrast, CDC6 exhibited a strong negative correlation with central memory CD4 T cells (cor =-0.75, P<0.001), while CHRNB2 was significantly negatively correlated with activated B cells (cor =-0.50, P<0.001). A total of 9 microRNAs (hsa-miR-26a-5p) and 31 long non-coding RNAs (lncRNAs) (GAS5) associated with biomarkers were predicted. Patients with high ADCY4 expression demonstrated a higher half-maximal inhibitory concentration (IC₅₀) value for gemcitabine compared to those with low expression. However, the opposite trend was observed for CDC6 and CHRNB2.<h4>Conclusions</h4>This study identified CHRNB2, ADCY4, and CDC6 as candidate biomarkers and provided preliminary clues for further exploration of potential therapeutic strategies for NPC.

<b>Background:</b> Among natural products, vanillin (<b>Van</b>), a major component of <i>Vanilla planifolia</i>, exhibits multiple bioactivities, including antimicrobial effects. <b>Methods:</b> In this study, <b>Van</b>, its analogues <i>o</i>-vanillin (<b>oVan</b>), <i>iso</i>-vanillin (<b>iVan</b>), ethylvanillin (<b>eVan</b>), and a library of newly synthesized derivatives were evaluated against <i>Helicobacter pylori</i> strains with distinct antibiotic susceptibilities. Time-kill kinetics, antibacterial spectrum, and viability in a normal gastric cell line GES-1, were also assessed. <b>Results: Van</b> showed minimal or no activity (MIC and MBC > 128 µg/mL), whereas structural modifications markedly improved anti-<i>H. pylori</i> activity, with MIC values as low as 4 µg/mL. Compounds <b>16V</b>, <b>20oV,</b> and <b>29eV</b> were among the most potent (MIC₉₀ = 4-16 µg/mL). Activity depended on both the vanilloid core and substituent type. The compounds were inactive against representative Gram-negative and Gram-positive bacteria (MIC > 128 µg/mL). Selected compounds preserved viability in GES-1 cells. Hierarchical clustering, artificial neural clustering, and principal component analysis identified potency-related architectural motifs and strain-specific activity. Docking against <i>H. pylori</i> urease suggested that several compounds, particularly <b>16V</b>, may interact with the enzyme, providing preliminary support for a possible involvement of this target. <b>Conclusions:</b> Systematic modification of the vanilloid scaffold generated selective and relatively non-cytotoxic anti-<i>H. pylori</i> hit compounds and confirmed the value of natural metabolites in antibacterial drug discovery.

The Huntingtin gene ( HTT ) contains a conserved, yet expandable CAG repeat within exon 1. While the pathogenic expansion in Huntington’s Disease (HD) is well studied, the role of surrounding domains remains unclear. Using genome-edited mini-organoids and neurons, we dissected HTT exon 1 and found species-specific toxicity: the human variant caused more severe deficits than the mouse. Swapping the proline-rich domain (PRD) - the most divergent region - revealed its key role: the mouse PRD mitigated, while the human PRD worsened neuronal phenotypes. Omics profiling showed that pathogenic human exon 1 induced broad protein dysregulation, largely reversed by mouse PRD replacement. Bioinformatics implicated the actin cytoskeleton and transcriptional coactivator MKL2/MRTFB. We validated MKL2/MRTFB dysregulation in HD models and showed that restoring its expression rescued neuronal abnormalities. These findings highlight the PRD’s contribution to HD toxicity and point to MKL2/MRTFB and the cytoskeleton as candidate mediators.

Oxidative stress is a well-established driver in the pathogenesis and progression of various kidney diseases. The peroxiredoxin (PRDXs) family, comprising typical 2-Cys, atypical 2-Cys, and 1-Cys members, serves as essential thiol-dependent peroxidases that maintain cellular redox balance. Unlike classical antioxidant enzymes that solely scavenge reactive oxygen species, PRDXs are unique in their ability to undergo redox-sensitive structural transitions, functioning as molecular chaperones and intracellular signaling hubs. In the kidney, PRDXs exert multifaceted protective roles by preserving mitochondrial integrity and attenuating inflammatory and fibrotic signaling. Interestingly, beyond these canonical functions, several PRDX members exhibit context-dependent detrimental effects that paradoxically aggravate renal injury. Such dual functionality is regulated through sophisticated mechanisms, including specific post-translational modifications, molecular chaperone switching, and extracellular release as damage-associated molecular patterns. Although PRDXs represent promising therapeutic targets and biomarkers, their functional duality poses considerable challenges for drug development. Future efforts must focus on spatiotemporally precise regulation to selectively augment PRDXs-mediated defense while mitigating their injury-promoting effects.

Leukodystrophies (LDs), a group of heterogeneous genetic disorders, are characterized by selective involvement of cerebral white matter, including abnormal white matter development and/or progressive degeneration. Oligodendrocytes, astrocytes, microglia, axons, and the neurovascular unit collectively contribute to white matter homeostasis and disease progression. Recently, genomic sequencing has identified pathogenic variants in the alanyl-tRNA synthetase 1 (<i>AARS1</i>) and alanyl-tRNA synthetase 2, mitochondrial (<i>AARS2</i>) genes in LD-related phenotypes. Dysfunction of AARS1 and AARS2 proteins may impair cytosolic or mitochondrial tRNA aminoacylation, compromise editing fidelity, and disrupt mitochondrial homeostasis, which may lead to disruption of protein homeostasis, cellular stress responses, and energy failure. Alanyl-tRNA synthetase (AlaRS) impairments play an important role in the pathological processes of cytosolic and mitochondrial alanyl-tRNA synthetase-related disorders. These molecular defects are associated with characteristic neuroimaging patterns and diverse clinical manifestations observed in <i>AARS1</i>/<i>AARS2</i>-related disorders. This review summarizes current knowledge on the genetic basis, clinicopathological features, and molecular mechanisms of <i>AARS1</i>- and <i>AARS2</i>-related leukodystrophies, and discusses emerging therapeutic perspectives, with the aim of facilitating precision diagnosis and future targeted interventions.

Artificial Intelligence (AI) has been increasingly applied to investigate genetic irregularities associated with Alzheimer's Disease (AD). However, its potential to uncover deeper, more detailed molecular and cellular mechanisms remains underexplored, primarily due to limitations in integrating large-scale data and capturing the complex, cell-type-specific dynamics involved in AD pathology. Single-cell RNA sequencing (scRNA-seq) has emerged as a powerful tool in transcriptomics, offering high-resolution, cell-specific insights into complex biological systems. Despite this advancement, a significant gap remains in identifying both common and cell-type-specific transcriptomic signatures that define AD-related cellular and molecular processes. To address this, we propose a deep learning framework leveraging a Multi-Layer Perceptron (MLP) to classify AD versus control nuclei using scRNA-seq data from the Religious Orders Study/Memory and Aging Project (ROSMAP). We focus on microglial subclusters, particularly those representing homeostatic and activated states, to train the MLP model for optimal classification performance. We utilize the predicted embeddings from the MLP to model a disease progression trajectory for each of the datasets. Our model demonstrates strong performance in both classification and disease trajectory inference. To enhance interpretability, SHapley Additive exPlanations (SHAP) are applied to identify key AD-associated genes. Based on the most salient genes implicated in AD, we built transcription gene regulatory networks, revealing novel transcription factors and regulons for AD pathogenesis. These regulons highlight profound impacts of dysregulations of proteostasis, endoplasmic reticulum (ER) stress responses, and circadian rhythm on synaptic plasticity and neuronal survival in AD, offering a more holistic approach to drug target discovery compared to conventional single-target strategies, potentially leading to greater efficacy in slowing or reversing disease progression. This work demonstrates the transformative potential of AI in elucidating the molecular mechanisms of AD, offering improvements over traditional methods and uncovering novel insights into disease pathogenesis and potential therapeutic targets.

<h4>ABSTRACT</h4> Huntington’s disease is a severe neurodegenerative condition arising from an abnormal CAG repeat expansion in the HTT gene, which leads to the production of a mutant Huntingtin protein carrying an extended polyglutamine stretch. Although the field has largely centred on the toxic effects gained by this mutant protein, growing evidence points to the loss of normal wild-type Huntingtin function as an additional contributor to disease progression. Despite this, the cell-intrinsic roles of wild-type Huntingtin in neuronal biology remain poorly defined, in part because disentangling its specific contributions from broader network-level effects has proven technically challenging. To address this knowledge gap, we took advantage of the Drosophila huntingtin homolog ( htt ) and selectively manipulated its expression in the small lateral ventral neurons (sLNvs), a discrete cluster of just eight circadian pacemaker neurons that govern behavioral rhythmicity and sleep. Through targeted genetic knock-down, we show that reducing htt levels in sLNvs weakens the robustness of free-running circadian rhythms and substantially increases sleep in female flies. These behavioral changes are not rooted in developmental abnormalities, as restricting htt knock-down to adult flies reproduces the sleep phenotype across both beam-crossing and video-based locomotion assays. At the cellular level, htt loss disrupts dense core vesicle trafficking along sLNv axons, altering the fraction of motile vesicles and their velocity, and abolishing the time-of-day-dependent fluctuations in vesicle dynamics observed in these neurons. Complementary electrophysiological recordings using whole-cell patch-clamp further reveal that htt knock-down lowers action potential firing rates without perturbing resting membrane potential. Together, these results identify huntingtin as a cell-autonomous regulator of neuropeptide trafficking, neuronal excitability and circadian output. Beyond advancing our understanding of wild-type huntingtin physiology, this work carries direct relevance for HD therapeutic strategies, particularly those involving huntingtin-lowering approaches, by highlighting functions that may be unintentionally compromised.

Stem cells are critical for the development and maintenance of tissue integrity. An important example is intestinal stem cells (ISCs) that generate all epithelial cell types necessary for formation of the intestinal lining. HAT1 is a histone acetyltransferase that acetylates newly synthesized histone H4 molecules on lysine residues 5 and 12 during replication-coupled chromatin assembly. Within the intestine, HAT1 is specifically expressed in intestinal stem and progenitor cells. We generated an inducible deletion of the Hat1 gene in intestinal epithelial cells. Following loss of Hat1, intestinal crypts became elongated, with an increase in stem and progenitor cell proliferation and an increase in the population of OLFM+ cells. Loss of Hat1 also resulted in alterations in ISC differentiation, including an increase in the number of Goblet cells and the mislocalization of Paneth cells into villi. HAT1 is specifically responsible for the acetylation of histone H4 lysine 5 (H4K5ac) in ISCs. Genome-wide characterization of HAT1-dependent H4K5ac in intestinal crypt cells indicates that the most significant loss of H4K5ac occurs regions of the genome that correspond to in lamina-associated domains, as defined in mouse embryonic fibroblasts. Loss of H4K5ac is accompanied by an increase in histone H3 K9 tri-methylation, indicating that HAT1 regulates genome-wide histone modification patterns in intestinal crypt cells. A direct role for HAT1 in ISC function was demonstrated using organoids in culture. HAT1 is required for differentiation in organoids and for the maintenance of Lgr5+ stem cells. These results indicate that HAT1 is required for the proper regulation of ISC renewal and differentiation.

Improving adipose tissue (AT) metabolism through exercise might increase production of lipokines from the fatty acid esters of hydroxy fatty acids (FAHFA) family. Although exercise training can increase insulin sensitivity and levels of specific FAHFA in AT and the circulation in humans, the effects of acute exercise on FAHFA levels remain unclear. Previously untrained lean, obese and aged male C57BL/6J mice were subjected to acute treadmill exercise. AT lipolysis, metabolomic profile and FAHFA levels were analysed at baseline and 0 and 3 h postexercise. Baseline AT levels of FAHFA, particularly those containing mono- and polyunsaturated fatty acids, were lower in obese and aged mice compared with lean mice. AT-specific expression of genes related to de novo lipogenesis and lipolysis was reduced, particularly in obese mice, whereas expression of FAHFA hydrolysis-related genes was higher in aged mice. In lean mice, acute exercise increased plasma non-esterified fatty acids and their release from AT explants, which was suppressed in obese and aged counterparts. Only in lean animals, acute exercise led to a time-dependent increase in total FAHFA levels in AT, with the most pronounced increase observed in 13/12-regioisomers from different FAHFA subfamilies. The effect of acute exercise on AT FAHFA levels was stronger than that of fasting alone, revealing palmitoleic acid-containing FAHFA as the most differentiating regioisomers. In contrast, plasma FAHFA levels were minimally affected in all groups. In conclusion, acute exercise increases FAHFA levels in AT but not in plasma, and this effect appears to be negatively affected by obesity and ageing.

Acute exposure to high-altitude hypobaric hypoxia (HH) disrupts physiological homeostasis. While gastrointestinal and neurological disturbances are its main manifestations, muscle function is also affected. However, its short-term effects on the gut-muscle axis remain poorly understood. The present study investigated the impact of a 4-hour exposure to simulated altitude (5,000 m) on intestinal morphology, gut microbiota composition, short-chain fatty acid (SCFA) levels, and skeletal muscle mitochondrial function in male Sprague-Dawley rats. Despite the severity of the hypoxic stimulus, biometric and hematological parameters remained largely unchanged, suggesting preserved systemic stability. Histological analysis revealed a significant reduction in intestinal crypt depth, indicating early structural alterations, while villus morphology and goblet cell counts remained unchanged. Microbiota profiling showed a decrease in Enterobacteriales abundance, while other bacterial groups and SCFA concentrations remained stable, pointing to limited microbial shifts under acute conditions. In skeletal muscle, reactive oxygen species (ROS) production increased under complex I-supported respiration while mitochondrial respiration was maintained across all respiratory states. This suggests a qualitative shift in redox balance. Collectively, these findings indicate that acute HH triggers early localized responses in intestinal and muscular tissues without inducing widespread systemic disruption, highlighting the resilience of the gut-muscle axis to short-term hypoxic stress. Longer or repeated exposures may be required to elicit broader physiological adaptations. This study contributes to understanding the early effects of altitude-related hypoxia and underscore the importance of exposure duration in shaping host responses.

<h4>Background</h4>Non-typeable <i>Streptococcus pneumoniae</i> (NT-Sp) strains lack detectable capsules and are often missed by traditional serotyping. While associated primarily with asymptomatic carriage, NT-Sp has emerged as a key player in antimicrobial resistance (AMR) gene dissemination and serotype replacement in the post-vaccine era. This systematic review and meta-analysis synthesize global evidence on NT-Sp carriage, its prevalence, antimicrobial resistance patterns, and genetic diversity.<h4>Methods</h4>A systematic search of PubMed, Scopus, Web of Science, Google Scholar, and ScienceDirect was conducted up to May 2025. Eligible studies reporting NT-Sp carriage, molecular typing, or AMR profiles were included. Data extraction and quality assessment followed PRISMA guidelines. A meta-analysis was performed to estimate pooled prevalence of NT-Sp carriage and resistance, with subgroup and meta-regression analyses exploring heterogeneity. Methodological quality was assessed using the Newcastle-Ottawa Scale adapted for cross-sectional studies. Only open-access and free full-text articles were included, which may have introduced selection bias.<h4>Results</h4>Forty studies from 23 countries were included. The pooled NT-Sp carriage prevalence was 4.64% (95% CI: 2.78%-6.92%), with substantial heterogeneity (<i>I</i> ² = 97.1%, 95% CI: 96.6%-97.5%). Subgroup analyses revealed geographic variation, with Oceania showing the highest prevalence (7.24%, 95% CI: 1.39%-17.04%) and North America the lowest (0.44%, 95% CI: 0.05%-1.11%). Age-stratified analysis showed that studies, including all ages had the highest estimate (29.88%, 95% CI: 0.00%-100.00%), while children had a pooled prevalence of 4.05% (95% CI: 0.00%-15.80%). Vaccination era did not significantly influence NT-Sp carriage (<i>p</i> = 0.312). Resistance was highest for co-trimoxazole (67.21%, 95% CI: 49.92%-82.73%), penicillin (44.57%, 95% CI: 24.45%-65.50%), and erythromycin (39.85%, 95% CI: 19.58%-61.77%). Meta-regression identified the "all ages" participant category as a significant predictor of lower NT-Sp prevalence (<i>p</i> = 0.034). Molecular tools revealed substantial genomic diversity, with dominant clonal complexes varying by region.<h4>Conclusion</h4>NT-Sp is a globally distributed and genetically diverse pneumococcal subset with considerable AMR potential. Its persistence post-PCV introduction and resistance gene carriage highlights the need for improved detection, molecular surveillance, and consideration in next-generation vaccine development. Specifically, clinical laboratories in high-prevalence settings should adopt molecular methods (e.g., PCR or WGS) to detect NT-Sp in carriage surveillance; empirical antibiotic guidelines for pediatric respiratory infections should consider NT-Sp resistance patterns, particularly the high prevalence of co-trimoxazole resistance (67%) and next-generation PCV trials should include NT-Sp carriage as an exploratory endpoint. Integrating NT-Sp into global pneumococcal control strategies is essential.

Progressive Familial Intrahepatic Cholestasis (PFIC) is a rare liver disorder that, although typically present in childhood, can also occur in adulthood. Early diagnosis is crucial for appropriate clinical management, prognostic assessment, and therapeutic decision-making; however, it remains challenging due to phenotypic variability and the frequent identification of genetic variants of uncertain significance (VUS). Advances in next-generation sequencing (NGS) and whole-exome sequencing (WES) have improved the detection of disease-associated variants, revealing that the same genetic variants-often in a heterozygous state-may lead to adult-onset cholestatic disease, with milder or atypical presentations, while still conferring a significant risk of progressive liver injury and related complications. To assess the diagnostic yield and clinical utility of NGS panels and WES in adults with suspected PFIC or unexplained cholestatic liver disease, focusing on variant interpretation, emerging disease-associated genes, and clinical significance of VUS. A literature review was conducted to assess molecular diagnostics in adult cryptogenic cholestasis, with a focus on studies employing targeted NGS panels and WES. Selected studies were examined for diagnostic yield, variant classification, interpretive challenges, including VUS, and integration with clinical data. Data on patient demographics, sequencing techniques, and variant interpretation strategies were extracted to evaluate the current capabilities and limitations of genomic testing. From an initial search of 211 publications, 15 studies met the inclusion criteria, comprising five large adult cohorts and ten single-patient or trio-based reports. Across 72 patients, sequencing technologies identified 75 pathogenic or likely pathogenic variants, predominantly missense mutations, demonstrating high genetic heterogeneity. The most implicated genes included <i>ABCB4</i>, <i>ABCB11</i>, <i>ATP8B1</i>, <i>TJP2</i>, and <i>USP53</i>, with diagnostic yields ranging from 13.2% in large heterogeneous cohorts to substantially higher rates in carefully selected individual cases. VUS were identified in up to 67% of cases, highlighting the need for multidisciplinary interpretation integrating clinical, biochemical, and genetic data. Emerging evidence also implicated ciliopathy-associated genes such as <i>DCDC2</i>, <i>NPHP3</i>, <i>PKHD1</i>, <i>TULP3</i>, and <i>TTC21B</i>, expanding the genetic spectrum of adult cholestasis. Clinically, presentations ranged from mild biochemical abnormalities to progressive cholestasis, with pruritus being a common feature across studies. Comprehensive genetic testing significantly enhances diagnostic accuracy, informs prognosis, and guides individualized management in adults with cryptogenic cholestasis. Emerging evidence suggests that ciliopathy-associated genes may contribute to the genetic architecture of adult cholestatic disorders, further expanding the spectrum of disease-associated genes. However, the high prevalence of VUS remains a major challenge, highlighting the need for functional studies, longitudinal follow-up, and improved variant interpretation frameworks. As genomic technologies and analytical approaches continue to evolve, genetic testing is expected to play an increasingly central role in precision hepatology.

Ferroptosis is an iron-dependent form of regulated cell death characterized by iron dyshomeostasis, glutathione depletion, glutathione peroxidase 4 dysfunction, and excessive lipid peroxidation. Exercise is a safe and accessible non-pharmacological intervention with broad neuroprotective potential, but the evidentiary basis linking exercise specifically to ferroptosis is uneven. Only a limited subset of studies directly combines an exercise intervention with ferroptosis-related outcomes in neurodegenerative models; much of the proposed pathway architecture is inferred from pharmacological, cellular, observational, or acute neurological injury studies. This review therefore separates direct exercise evidence from exercise-related supporting evidence and non-exercise mechanistic evidence. The most directly relevant findings, concentrated largely in aerobic exercise models, show exercise-associated changes in brain iron handling, the cystine/glutamate antiporter-glutathione peroxidase 4 antioxidant system, and lipoxygenase-dependent lipid peroxidation. Supporting studies suggest additional peripheral-to-central mechanisms involving muscle-derived exosomes, exercise-associated changes in systemic and cerebral iron handling, and inflammatory regulation. Bone marrow hematopoiesis, adult neurogenesis, synaptic plasticity, and astrocyte-controlled iron traffic are incorporated as biologically plausible but incompletely tested links. Evidence for resistance training, high-intensity interval training, mind-body exercise, and human disease remains insufficient. The central limitation is therefore not pathway plausibility but the scarcity of exercise-specific causal experiments demonstrating that ferroptosis suppression is required for neuroprotection.

Distant metastasis, characterized by organotropism, is a major cause of mortality in lung adenocarcinoma (LUAD). In this study, digital spatial profiling (DSP), multiplex immunofluorescence (mIF), and clinical data from 52 LUAD patients were integrated to develop organ-specific metastasis risk models, and the molecular mechanisms underlying metastatic organotropism were investigated. Random forest models based on primary tumor spatial transcriptomics accurately predicted metastasis to the brain (AUC = 0.974), liver (AUC = 0.975), adrenal gland (AUC = 0.929), and bone (AUC = 0.907). Key compartment-specific gene expression signatures associated with organotropic metastasis were identified, including those expressed in tumors (e.g., FKBP1A for the brain and MOCOS for the liver), immune cells (e.g., ADAMTSL2 for the liver), and stromal cells (e.g., CKAP2 for the brain). Pathway analyses revealed distinct biological processes associated with organotropism, such as enriched cell death pathways in brain metastasis and extracellular matrix (ECM) remodeling in liver metastasis. Postmetastasis survival models highlight stromal gene expression (e.g., PKM for OS and VCAM1 for PFS) and immunosuppressive microenvironments (e.g., M2 macrophage infiltration) as critical prognostic factors. The high-precision prediction models and key molecular signatures identified in this study enhance our understanding of "seed-soil" interaction dynamics and offer promising biomarkers and therapeutic targets for future clinical use.

The signaling mechanisms and developmental dynamics that govern the divergence of myocardial and epicardial lineages during human heart development remain poorly understood. Here, we developed a human pluripotent stem cell-based cardiac development model and employed time-course single-cell RNA sequencing to delineate cardiac lineage specification trajectories. We identified retinoic acid (RA) as a critical fate switch at the cardiac mesoderm stage. RA instructs epicardial lineage commitment of cardiac mesoderm through a primed-epicardium to proepicardium-like population and finally to epicardium, a process requiring precise BMP modulation. Conversely, RA absence directs cardiac mesoderm along a default myocardial pathway, yielding developing and mature cardiomyocytes. Both trajectories are governed by the hierarchical activation of key transcription factors. Our study integrates signaling and dynamics to elucidate the temporal regulatory network of the RA-BMP axis in human cardiac fate determination. These findings provide fundamental insights into human cardiogenesis and a crucial roadmap for modeling heart disease and advancing regenerative strategies.

Microarray and Next Generation Sequencing studies offer insight into gene regulation in Parkinson's disease (PD). However, analysing vast numbers of genes can make the interpretation of data difficult when considering the platforms and techniques used across different studies. In this study, transcript expression, restricted to genes related to G-protein coupled receptors, activating agonists, agonist precursors, synthesis enzymes and transduction processes, as well as stress-related chaperones and solute carriers, was assessed across nine microarray platforms and two RNAseq studies of the substantia nigra (nigral volume). Changes in gene expression associated with PD were assessed by differential expression analysis while RNAseq studies were also used to calculate transcript per million values for each of the genes within the nigral volume. This analysis showed extensive changes in nigral volume signalling for several robustly expressed signal-related transcripts including tyrosine hydroxylase (TH), WNT signalling components (SFRP1, RSPO2 and DKK3), Kallikrein Related Peptidase 6 (KLK6), neurexins (NRXN1, NRXN3), prostaglandin synthases (PTGES2, PTGES3) and fractalkine (CX3CL1). The nigral volume signalling ligand data was then cross-referenced to the most abundant G-protein coupled receptor and signal transduction transcripts in the pigmented neurons using two RNAseq and two microarray studies. There were 32 significant changes in G-protein coupled receptor and associated signalling genes in pigmented neurons from PD tissue. Of these, 30 genes were upregulated. Analysis of the transcription factors likely regulating the 30 upregulated genes indicates a stimulation of cell stress pathways, particularly the JNK-MAP kinase pathway. Following this pathway back to changes in nigral signalling transcripts indicates that deficits in at least two autocrine/paracrine signalling systems; dopamine, via Gαo-coupled dopamine D2 receptors and Dickkopf-3 (DKK3) appear likely to contribute to pigmented neuron stress in PD.

The OX40-OX40L co-stimulatory pathway has emerged as an important regulator of T-cell persistence and immune memory. While traditionally associated with type 2 inflammation, its role extends across multiple barrier tissues, including the skin and respiratory mucosa, where it integrates epithelial signals with adaptive immune responses. To provide a comprehensive and integrative overview of the OX40-OX40L axis as a shared driver of immune persistence across skin and airway inflammatory diseases and to discuss its therapeutic implications. A narrative review was conducted based on a focused literature search of PubMed, Scopus, and Web of Science up to February 2026. Experimental, translational, and clinical studies addressing OX40-OX40L biology and therapeutic targeting were included. OX40 signaling acts as a late co-stimulatory pathway that sustains effector T-cell survival, promotes tissue-resident memory T-cell formation, and modulates regulatory T-cell function. In barrier tissues, epithelial-derived alarmins such as TSLP and IL-33 induce OX40L expression on antigen-presenting cells, reinforcing type 2-polarized immune responses. This axis contributes to chronic inflammation in atopic dermatitis, asthma, allergic rhinitis, and nasal polyposis, supporting the concept of shared mechanisms of immune persistence across type 2 inflammatory diseases. Beyond type 2 immunity, OX40-OX40L signaling also participates in Th1-, Th17-, and Th22-mediated responses, as well as in autoimmune, fibrosing, and lymphoproliferative disorders. Therapeutic targeting of this pathway, particularly with monoclonal antibodies such as amlitelimab, has demonstrated clinically meaningful efficacy and suggests potential for durable disease modification. The OX40-OX40L axis represents an important immunological pathway linking epithelial activation to adaptive immune memory across barrier tissues. Its role in sustaining immune persistence provides a conceptual framework for understanding chronic inflammatory diseases and supports continued investigation of therapies targeting upstream co-stimulatory pathways.

Obsessive-compulsive disorder (OCD) remains difficult to treat in a substantial minority of patients, despite the established first-line use of exposure and response prevention, selective serotonin reuptake inhibitors, and pharmacological augmentation. This treatment gap has encouraged renewed interest in biological axes that sit outside a narrow monoamine-centered model. In this study, we asked whether transcriptional genetic signals for OCD show enrichment in cellular maintenance pathways motivated by nicotinamide mononucleotide (NMN)-associated aging biology and whether any such pathways outperform a curated antidepressant-related gene comparator. Using OCD transcriptome-wide association study (TWAS)-derived gene-level Z statistics and an absolute Z-score ranking framework, ten NMN-motivated Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway expansions were compared against AntiDep_Genes. The strongest and most defensible finding in the OCD analysis was selective enrichment of KEGG_AUTOPHAGY_ANIMAL. Autophagy nominally outperformed the antidepressant comparator in the primary head-to-head test, with normalized enrichment score (NES) = 1.166, p = 0.041979, and difference in normalized enrichment score (dNES) = +0.2097. The signal persisted after removal of overlapping antidepressant-related genes, with cleaned NES = 1.1784 and p = 0.042479, and the autophagy set also showed a higher absolute Z-score distribution than AntiDep_Genes by Mann-Whitney U testing, p = 0.04717. Leading-edge overlap with the antidepressant comparator was minimal, limited to mechanistic target of rapamycin (MTOR). The OCD autophagy signal was driven by a mixture of core autophagy machinery and upstream signaling genes, including MAPK3, MAP1LC3A, PPP2CB, MAP2K1, GORASP2, BAD, ATG2A, ATG10, MAPK8, and CALCOCO2. This pattern suggests a signaling-execution module rather than a simple enrichment of canonical ATG genes alone. The result is biologically consistent with recent OCD genetic findings implicating cortical and hippocampal excitatory neurons and D1/D2 striatal medium spiny neurons, which are cell types with high synaptic and metabolic demands. These findings are best viewed as hypothesis-generating. They do not support NMN, nicotinamide adenine dinucleotide (NAD+) precursors, or autophagy modulators as OCD treatments, but they do identify autophagy and mitochondrial quality-control biology as plausible targets for future functional and stratification studies. The autophagy enrichment was nominally significant in the primary analysis but did not survive Benjamini-Hochberg false-discovery rate correction across the ten tested pathways (false discovery rate (FDR)-adjusted p = 0.41979); the findings should therefore be considered hypothesis-generating.

<h4>Introduction</h4>The present study analyzed the specific function of MARVEL domaincontaining protein 3 (MARVELD3) in esophageal cancer (EC).<h4>Methods</h4>MARVELD3 expression and its association with clinicopathological features in EC samples were analyzed using data from the TCGA and GEO databases. Enrichment analysis was performed on differentially expressed genes (DEGs) in samples with high and low MARVELD3 expression. Furthermore, the relationships of MARVELD3 with immune infiltration, response to immune therapy, N6-methyladenosine (m6A) modification, cuproptosis, and ceRNA regulatory network were explored.<h4>Results</h4>In EC, MARVELD3 exhibited high expression, which was significantly correlated with the T-stage and histological type. ROC curve analysis demonstrated a high prediction accuracy of MARVELD3 for tumor prognosis. Moreover, immune infiltration analysis showed a close relationship between MARVELD3 and the immune microenvironment and immune cells in EC. Additionally, MARVELD3, the m6A modification-related gene YTHDF1, and the cuproptosis-related gene CDK5RAP1 were closely correlated. Furthermore, the predicted ceRNA regulatory network THRBIT1/ hsa-miR-23a-3p/MARVELD3 and LINC00261/hsa-miR-23a-3p/MARVELD3 axes may be involved in the pathogenesis of EC and its progression.<h4>Discussion</h4>This study analyzed the involvement of MARVELD3 in EC and revealed its relationship with m6A modification and cuproptosis and its relevant ceRNA regulatory network.<h4>Conclusion</h4>MARVELD3 may be a potential biomarker for EC and was closely linked to immune infiltration, response to immune therapy, m6A modification, cuproptosis, and ceRNA regulatory networks in EC.

Vaccination relies on innate and adaptive responses initiated at the injection site followed by activation in draining lymph nodes. However, high-plex, spatially resolved maps that couple injection site signals with transcriptomic programs in draining lymph nodes over time are still lacking. Here, we used spatial transcriptomics to map early responses to an adjuvanted recombinant haemagglutinin (rHA) protein vaccine formulated with Sanofi's proprietary oil-in-water adjuvant AF03. Mice received intramuscular rHA-protein or PBS. Injection site muscle and draining lymph nodes were collected at Days 1 and 7 post-prime for GeoMx™ Digital Spatial Profiling (DSP) and CosMx™ Spatial Molecular Imaging (SMI) single-cell imaging. In lymph nodes, rHA-protein vaccination induced early innate pathway activation in non-follicular regions at Day 1. By Day 7, adaptive pathways were enriched within follicles, with differentially expressed genes (DEGs) linked to germinal center (GC) formation, transcriptional regulation and signaling. These changes coincided with expansion of a proliferative GC-like B-cell cluster within the follicular area. At the injection site, 20x20 µM spatial grids were used rather than segmented cells. Clusters, enriched for fibroblast and muscle cells showed Day 1 induction of chemokines and interferon stimulated genes, concentrated in fibroblast rich stromal layers. These spatially organized tissue programs were accompanied by transient systemic cytokine elevations and increased antigen-specific IgG titers. Together, these data provide a previously unresolved view of what, when and where local inflammation and lymph node organization are spatially and temporally associated during early responses following vaccination with an adjuvanted rHA-protein vaccine.

<h4>Objective</h4>To investigate whether Ginkgolide B (GB) protects retinal pigment epithelial cells (RPEs) from hydrogen peroxide (H₂O₂)-induced oxidative stress and apoptosis, and to explore the potential involvement of the adenosine monophosphate-activated protein kinase <i>α</i> (AMPKα)/nuclear factor erythroid 2-related factor 2 (Nrf2) pathway.<h4>Methods</h4>Human RPEs were divided into the following groups: control, model (200 μmol/L H₂O₂ for 24 h), GB + H₂O₂, GB + H₂O₂ + Compound C (an AMPK inhibitor), H₂O₂ + Compound C, and GB alone. Cell viability, apoptosis, antioxidant parameters, and AMPKα/Nrf2 expression were assessed using the Cell Counting Kit-8 (CCK-8) assay, flow cytometry, colorimetric biochemical assays, transmission electron microscopy (TEM), quantitative polymerase chain reaction (PCR), and Western blot.<h4>Results</h4>TEM showed that GB attenuated H₂O₂-induced mitochondrial swelling and cristae disruption, which were reversed by Compound C. Compared with the model group, the GB + H₂O₂ group exhibited increased cell viability, decreased apoptosis, elevated activities of catalase (CAT), glutathione peroxidase (GPx), and superoxide dismutase (SOD), as well as increased glutathione (GSH) content, and reduced malondialdehyde (MDA) (all <i>p</i> < 0.05), though protein carbonyl (PC) levels showed no significant amelioration. All protective effects were attenuated by Compound C. In the GB-alone group, the percentage of apoptotic cells, Nrf2 mRNA expression, and all other parameters did not differ significantly from those in the control group.<h4>Conclusion</h4>GB was associated with protection of RPEs against H₂O₂-induced oxidative stress and apoptosis. Because these protective effects were attenuated by pharmacological inhibition of AMPK, the findings suggest that AMPK/Nrf2 signaling may contribute to the cytoprotective actions of GB. However, additional mechanistic studies are required to confirm the involvement of this pathway.

Caspase-8 and the NF-κB essential modulator (NEMO, also referred to as IKKγ) play critical roles in controlling TNF-α-induced cell death and survival in hepatocytes. The aim of this study was to generate a hepatocyte-derived cell line in which the <i>Casp8</i> and <i>Nemo</i> genes can be conditionally inactivated simultaneously to investigate the significance of the corresponding signaling pathways. To this end, we induced hepatocellular carcinoma in <i>Casp8</i>^f/f<i>Nemo</i>^f/f mice using diethylnitrosamine and established an immortalized hepatoma cell line from explanted liver tumors, which is subsequently referred to as ΔCN60. ΔCN60 cells still retain floxed <i>Casp8</i> and <i>Nemo</i> alleles, allowing for efficient Cre-mediated deletion to generate <i>Casp8</i>^Δ<i>Nemo</i>^Δ derivatives. Loss of both Caspase-8 and NEMO inhibits cell proliferation, increases the expression of tumor and progenitor markers (AFP, CD133), reduces albumin expression, and blocks TNF-α-induced NF-κB p65 nuclear translocation. ΔCN60 <i>Casp8^ΔNemo^Δ</i> cells display altered sensitivity to prolonged TNF-α exposure, suggesting a potential shift in necroptosis-associated signaling pathways, although necroptotic cell death was not directly demonstrated. ΔCN60 serves as a versatile hepatoma model for investigating Caspase-8/NEMO-dependent TNF-α signaling and hepatocyte plasticity and may help prioritize future in vivo experiments by enabling selected mechanistic questions to be addressed first in vitro.

This review covers published data (mostly from 2019 to 2025) on the synthesis and biological activity of azolo[<i>a</i>]quinoxalines, including pyrazolo-, imidazo- and triazolo-annelated systems. We highlight that most research efforts are directed toward the design of anticancer agents, with additional applications as Toll-like receptor antagonists, monoamine oxidase inhibitors, opioid receptor modulators, PI3Kα inhibitors, tubulin polymerization inhibitors, GABAᴀ receptor modulators, VEGFR-2 kinase inhibitors, BRD9 binders, and anti-inflammatory, antimicrobial, and antifungal agents. Recent synthetic strategies include Cu-catalyzed oxidative annulations, I₂-mediated C-H functionalization, metal-free cascade cyclization, and multicomponent reactions, often employing eco-friendly catalysts and reductants. A growing number of studies integrate virtual screening, molecular docking, and pharmacophore-based in silico approaches to guide lead discovery and optimization. Innovative drug delivery systems, such as nanogels and hybrid molecules combining azoloquinoxalines with pharmacophores like thalidomide, have also been explored. This review emphasizes both the medicinal chemistry aspects of azolo[<i>a</i>]quinoxalines and the synthetic methodologies for their preparation from the perspective of drug development and discovery.

Aging is driven by progressive epigenetic alterations-DNA methylation drift, aberrant histone modifications, chromatin remodeling, and non-coding RNA dysregulation. Vitamin D, acting through its nuclear receptor vitamin D receptor (VDR), modulates the epigenetic landscape to potentially counteract these age-related changes. This review first describes age-related epigenetic alterations, then outlines vitamin D signaling and its interface with the epigenetic machinery. Next, tissue-specific epigenetic actions of vitamin D in the immune, musculoskeletal, and nervous systems are discussed. Finally, clinical trial evidence is examined, interindividual variability is highlighted, and future research directions are proposed. However, large randomized controlled trials (RCTs) consistently show limited benefits of vitamin D monotherapy, with measurable anti-aging effects observed when combined with exercise and nutritional interventions. Its efficacy is constrained by interindividual variability, J-shaped dose-response, and tissue-specific barriers. For deficient individuals (serum 25-hydroxyvitamin D (25(OH)D) < 50 nmol/L), guided supplementation-typically 800-2000 international units (IU)/day-is warranted to achieve tentative target serum concentrations of 75-125 nmol/L, the range linked to favorable epigenetic and immune effects. For those already sufficient (e.g., serum 25(OH)D ≥ 50 nmol/L), indiscriminate supplementation without biochemical indication is not supported. Therefore, promoting healthy aging through vitamin D requires serum-monitored, individually titrated, and multimodal regimens, with supplementation reserved primarily for documented deficiency and integrated with lifestyle interventions.

Histone-modifying enzymes and histone post-translational modifications (PTMs) play key roles in the organization (euchromatin versus heterochromatin) and function (active versus silenced genes) of chromatin. The abundance and activity of these enzymes, along with their associated histone PTMs, are often altered in cancer cells, leading to deregulated gene expression. The expression of the KMT2A-MLLT3 protein, resulting from a chromosomal translocation in mixed-lineage leukemia (MLL), a subtype of acute myeloid leukemia, augments transcription elongation, promoting the expression of <i>HOXA9</i> and <i>MEIS1</i>, genes that play critical roles in MLL development. Bortezomib, a proteasome inhibitor, has been effective at treating various cancers. In this study, we compared the impact of bortezomib on histone PTMs in the MLL cell line MOLM-13 and the chronic myeloid leukemic (CML) cell line K562. We report that MOLM-13 had a greater level of histone H2B monoubiquitinated at lysine 120 (H2BK120ub) and histone H3 dimethylated at lysine 79 (H3K79me2) (modifications involved in elongation) and similar levels of histone H2A monoubiquitinated at lysine 119 (H2AK119ub). Bortezomib treatment resulted in significant reductions in H2BK120ub and H2AK119ub levels, as well as in transcript levels of genes involved in MLL development.

<h4>Background</h4>Autoimmune diseases (AIDs) are associated with increased cardiovascular risk. However, specific protein mediators linking AIDs to major adverse cardiovascular events (MACE) and cardiovascular death (CV death) remain unexplored. This study identifies proteomic mediators linking AIDs to MACE via high-dimensional mediation analysis in the UK Biobank.<h4>Methods</h4>We used UK Biobank data with proteomic profiling by Olink platform. Participants with prevalent myocardial infarction (MI), stroke, and heart failure at baseline were excluded. AIDs were categorized into musculoskeletal (MSK), vasculitis, gastrointestinal (GI), neurologic, and rheumatic fever subsets. Fine-Gray models assessed associations between AIDs and MACE and CV death. Proteome-wide association studies identified proteins associated with both AIDs and cardiovascular outcomes. High-dimensional mediation analysis (HIMA) explored protein-mediated pathways. All models adjusted for age, sex, lipids, BMI, smoking, hypertension, diabetes, chronic kidney disease, atrial fibrillation, and coronary artery disease.<h4>Results</h4>Among 400,633 participants (median follow-up 14.5 years, 44.8% male), AIDs were present in 28,754 (7.2%). All AID categories were associated with increased MACE (sHR: MSK 1.34, vasculitis 1.67, GI 1.20, neurologic 1.33, rheumatic fever 1.38; all <i>p</i> < 0.001). For CV death, MSK, vasculitis, and rheumatic fever showed increased risk (sHR 1.34, 1.78, 1.51; all <i>p</i> ≤ 0.004), but not GI or neurologic AIDs. In 43,599 participants with proteomic data, HIMA identified 66 and 32 unique potential mediators linking AIDs to MACE and CV death, respectively. Four proteins (Growth Differentiation Factor 15, Interleukin-15, urokinase plasminogen activator receptor, and Tenascin C) mediated the AID-MACE relationship across multiple AID categories. Growth Differentiation Factor 15 and Interleukin-15 were shared mediators for CV death.<h4>Conclusions</h4>This proteomic analysis identifies specific proteins that may mediate the association between AIDs and adverse cardiovascular outcomes, offering mechanistic insights into immune-related cardiovascular risk. These findings are hypothesis-generating and require replication and validation before the identified proteins can be considered causal mediators or adopted for clinical risk stratification.

<b>Purpose:</b> Hypertension in women is a dynamic, hormone sensitive condition shaped by cumulative physiological changes across the life course. This review summarises current evidence relating vascular ageing, hormonal regulation, metabolic dysfunction, and reproductive history to blood pressure regulation in women. <b>Materials and Methods:</b> A narrative review of the literature was conducted using PubMed, Scopus and Google Scholar. Clinical, epidemiological, and mechanistic studies were synthesised to evaluate factors influencing hypertension in women. Reports in which menopausal status was not defined, or previous reproductive milestones were not documented, were excluded or interpreted with caution. <b>Results:</b> Evidence suggests that menopause, vascular ageing, metabolic dysfunction, androgen to oestrogen balance, and reproductive history interact to influence endothelial function, neurohormonal regulation, renal sodium handling, and vascular resistance. Ageing-related mechanisms, including cellular senescence, chronic low-grade inflammation, and genetic susceptibility, may contribute to increased cardiovascular risk. Hypertensive disorders of pregnancy identify women who are at higher risk of developing cardiovascular disease later in life and provide an opportunity for earlier risk assessment and prevention. Emerging therapies, including GLP-1 receptor agonists and SGLT2 inhibitors, may offer additional options for improving cardiovascular risk management, although their role in sex-specific prevention remains an evolving area of research. <b>Conclusions:</b> Hypertension in women is best understood within a life-course framework. Incorporating reproductive history, menopausal status, metabolic health, and emerging risk markers may improve cardiovascular risk assessment and support earlier intervention.

Peroxiredoxin 6 (PRDX6) is a multifunctional enzyme with peroxidase, calcium independent phospholipase A 2 (aiPLA₂) and lysophospatidylcholine acyltransterase (LPCAT) activities. Although PRDX6 can repair peroxidised phospholipids and help prevent ferroptosis, its role in the adaptive response to physiological oxidative stress remains unclear. In this study PRDX6 function was investigated using myoblasts exposed to an acute low concentration of H 2 O 2 and Caenorhabditis elegans subjected to a swimming intervention. Mild oxidative stress promoted myogenesis and mitochondrial turnover in myoblasts, while exercise enhanced activity and longevity in C. elegans . The adaptive responses were associated with increased mitochondrial localisation of PRDX6. In contrast, loss of PRDX6 under mild stress conditions resulted in increased mitochondrial lipid peroxidation, enhanced mitochondrial ER contact sites (MERCS), mitochondrial calcium accumulation, release of mitochondrial DNA and activation of innate immune signalling pathways. Similar phenotypes were induced by the ferroptosis activator erastin and rescued by the lipid peroxyl scavenger Ferrostatin-1, indicating lipid peroxidation was the key triggering event. Furthermore, inhibition of mitochondrial calcium uptake in C. elegans prevented calcium overload and attenuated inflammatory signalling. Together, the results identify PRDX6 as a conserved regulator of mitochondrial adaptation to physiological oxidative stress, functioning to limit mitochondrial lipid peroxidation and prevent excessive MERCS assembly, mitochondrial calcium dysregulation and inflammatory activation.

<h4>Background</h4>Primary cilia are essential for skeletal development by coordinating key signalling pathways in osteoblasts and chondrocytes. While pathogenic variants in approximately 40 genes have been linked to skeletal ciliopathies, additional causative genes remain to be identified. Although DNAH10 is well characterised in motile cilia, its role in primary cilia remains unclear.<h4>Methods</h4>Whole-exome sequencing was performed in two individuals with skeletal developmental abnormalities and identified biallelic <i>DNAH10</i> variants. In vitro assays were used to assess the effects of these variants on DNAH10 protein abundance. <i>Dnah10</i> knockout mice were generated to investigate the associated skeletal phenotypes and underlying mechanisms.<h4>Results</h4>Biallelic <i>DNAH10</i> variants were identified in two individuals with skeletal developmental abnormalities. In vitro, these missense variants were associated with reduced DNAH10 protein abundance. <i>Dnah10</i>-deficient mice recapitulated key skeletal features, including polydactyly, impaired cartilage development and abnormal ossification. Mechanistically, <i>Dnah10</i> loss was associated with abnormal primary cilia morphology, attenuated Hedgehog signalling, impaired osteoblast differentiation and defective chondrocyte maturation.<h4>Conclusion</h4>Our findings support <i>DNAH10</i> as a candidate gene associated with skeletal developmental abnormalities and ciliary dysfunction and suggest a previously underappreciated role for DNAH10 in primary cilia-related skeletal development beyond its established role in motile cilia.

<h4>Background</h4>The transcription process is controlled by non-coding regulatory elements, more than 70% of which are putative enhancers. These enhancers comprise over 600,000 regions and are marked by histone modifications. However, the mechanisms by which altered enhancers in cancer cooperate within the three-dimensional chromatin architecture to drive oncogenic programs remain poorly understood.<h4>Results</h4>By integrating 201 H3K27ac ChIP-seq datasets from prostate, we identify 3,216 high-confidence prostate cancer-specific putative enhancers. Ultra-high-resolution chromatin interaction profiling by Region Capture Micro-C at a representative chr6q24.1 locus reveals that these enhancers form cancer-specific, highly nested interactions with promoters that coalesce into a multi-connected hub absent in normal prostate cells. CRISPR/Cas9 perturbations of these enhancers, examined one by one, distinguish enhancer classes within the hub. Deletion of a central enhancer collapses hub-wide enhancer activities and architecture, leading to the downregulation of target genes, impaired proliferation, and reduced clonogenic growth. In contrast, deletion of a redundant enhancer results in minimal transcriptional changes, as neighboring enhancers rescue cancer signaling through compensatory architectural rewiring that strengthens alternative enhancer-promoter interactions. We also observe that FOXA1, a pioneer transcription factor activated in prostate cancer, directly binds to these enhancers and regulates distinct enhancer classes, leading to varying degrees of chromatin accessibility and gene expression changes.<h4>Conclusions</h4>These findings suggest that enhancers function in a coordinated manner, forming multi-connected cancer-specific chromatin interaction hubs, with distinct enhancer classes contributing differently to gene regulation. This study advances our ability to modulate gene expression in a cell type-specific manner, opening new avenues for precision therapies.

Ischemic stroke induces oxidative stress, neuroinflammation, neuronal death, and synaptic dysfunction, leading to persistent motor and cognitive deficits. The human dental pulp stem cell (hDPSC) secretome is a promising cell-free therapeutic candidate containing neurotrophic, antioxidant, and immunomodulatory factors. Here, we investigated its therapeutic effects in a photothrombotic mouse model of ischemic stroke and CoCl₂-induced hypoxic BV2 microglial cells. Proteomic profiling identified antioxidant-associated proteins, including SOD2, GSR, and GSTP1, and microglial phenotype-related candidates, including GRN, CSF1, and LRP1. hDPSC secretome treatment reduced stroke-induced infarct volume and attenuated stroke-increased neuronal apoptosis, neuronal ROS accumulation, and NF-κB-associated inflammatory signaling in the cortex and hippocampus. It also shifted microglial marker expression toward an M2-associated profile and improved stroke-impaired hippocampal neurogenesis, vascular remodeling, and synaptic organization. Proteomic analyses further identified coordinated changes in pathways related to oxidative phosphorylation, inflammatory responses, calcium signaling, SNARE-associated vesicular transport, and ROBO-Rho-associated cytoskeletal remodeling. These molecular and cellular changes were associated with improved motor coordination, spatial learning and memory, contextual memory, and anxiety-like behavior. These findings support the hDPSC secretome as a cell-free therapeutic candidate for post-stroke functional recovery linked to redox, inflammatory, neurovascular, and synaptic remodeling.

Transcription is spatially organized in the nucleus, concentrating transcription factors, coactivators, and RNA polymerase II into dynamic, membraneless compartments. Such assemblies have been described in many ways, from small complexes of defined stoichiometry to membraneless biomolecular condensates. We argue that these are not competing descriptions but positions along a single continuum, and propose "transcriptional compartment" as an umbrella term for it. We then turn to the functional literature and ask what these compartments have been shown to do, and what they have not. The clearest evidence for a functional role of condensation may emerge in contexts where transcription must generate switch-like responses, such as cell-fate decisions, stress responses, environmental sensing, and disease. Much of the remaining biology, however, sits in the small, transient assemblies between the two extremes. These are the hardest to study, in part because they fall below the diffraction limit of light microscopy and because the interactions that drive their formation usually also carry out their function, so the two cannot be perturbed independently. We survey the experimental and computational toolkit available for this problem and outline the open methodological challenges that remain.

Inflammatory osteolysis arises from pro-inflammatory cytokine-driven osteoclast activation and disrupted bone remodeling equilibrium. The IRE1-XBP1s axis, a major unfolded protein response pathway, regulates cellular homeostasis, but its role in inflammatory osteoclastogenesis remained unexplored. Single-cell RNA-seq showed increased osteoclast precursor cells and activated IRE1-XBP1s in LPS-induced osteolysis. Inhibition of this axis reduced osteoclastogenesis and bone loss in vitro and in vivo. RNA-seq indicated that blocking IRE1-XBP1s suppressed Slc6a4 transcription, with gene set enrichment analysis confirming its role in 5-HT transport. Dual-luciferase assays and ChIP-PCR demonstrated XBP1s' direct transcriptional regulation targeting the Slc6a4 promoter. The 5-HT transporter inhibitor escitalopram also inhibited osteoclastogenesis, highlighting the IRE1-XBP1s-Slc6a4 axis's importance. Notably, untargeted metabolomics suggested 5-HT inhibited intracellular 3-methyladenine (3MA) metabolism, a compound previously considered unnatural. HPLC-MS confirmed the presence of 3MA metabolism in inflammatory osteoclasts, and 3MA supplementation attenuated 5-HT-induced autophagy and osteoclast differentiation. Blocking the IRE1-XBP1s-Slc6a4 axis reduced pro-osteoclastogenic effects in inflammatory bone disease patient-derived PBMCs. This study demonstrates that IRE1-XBP1s inhibition alleviates inflammatory osteoclastogenesis and osteolysis via a 5-HT-dependent anti-autophagy mechanism, proposing this pathway as a therapeutic target for inflammatory bone loss.

Bromodomain-containing protein 4 (BRD4) is an important therapeutic target for anticancer, antiviral, and anti-inflammatory responses. Although its role in transcription, chromatin dynamics, and epigenetic programs is well recognized, recent studies have expanded the functions of BRD4 and its related bromodomain and extra-terminal (BET) family members (BRD2, BRD3, and BRDT) to other molecular and biological processes. How a universal epigenetic regulator is tailored for context-specific gene/pathway regulation is mechanistically intriguing. In this review, we highlight the importance of protein isoforms and posttranslational modifications, particularly phosphorylation, in generating the protein diversity necessary for selective factor recruitment and context-dependent regulation. Three BRD4 protein isoforms have been identified, including the universally expressed BRD4-L and BRD4-S(a) (simplified as BRD4-S), representing the long and short isoform a, and cell-/stress-specific short isoform b, BRD4-S(b). A phospho-switch mechanism controlling the open/closed state of the bromodomain and BRD4 interaction with partner proteins will also be discussed. Mechanistic understanding of BET protein action has led not only to the development of diverse bromodomain-binding compounds currently used in clinical trials but also to the discovery of a new class of small-molecule inhibitors targeting an intrinsically disordered region (IDR) of phospho-BRD4 to alter specific protein-protein interaction (PPI) networks without globally perturbing transcription programs and chromatin landscapes, thus significantly reducing off-target effects and providing new directions for therapeutic drug development.

Cancer arises from genetic and epigenetic alterations that reshape chromatin, transcriptional regulation, and malignant cell states. To chart cancer-intrinsic regulatory programs, we build a pan-cancer single-cell atlas of 60 cancer cell lines spanning 16 tissue origins and 20 cancer types, comprising 240,957 snRNA-seq and 223,347 snATAC-seq profiles. Integrative analyses reveal cell-state heterogeneity, core gene-regulatory networks, and a conserved EMT axis transcending tissue of origin; copy-number analysis identifies transcription factor amplification and hyperactivation as drivers of state reprogramming. Comparing cutaneous melanoma with acral melanoma, a rare subtype underrepresented in previous studies, uncovers a universal inflammation-suppressive program in acral and an inflamed landscape in cutaneous melanoma, with JAK-STAT activity as the central discriminator. Integrating data across models and patient cohorts links tumor-intrinsic regulation to microenvironmental composition and therapeutic response. By profiling rare alongside common subtypes, this atlas offers a resource for mapping pan-cancer and subtype-specific regulatory programs shaping cell-state plasticity.

In contrast to the ubiquitin (Ub)-proteasome-system, which only degrades individual proteins, macroautophagy can eliminate protein complexes or aggregates, organelles and even pathogens. Terms such as mitophagy, aggrephagy, lysophagy and xenophagy have been coined based on the targeted substrate. In Ub-dependent selective macroautophagy, cargo selectivity is specified by E3 Ub ligases that append Ub chains that in turn are recognized by selective autophagy receptors (SARs), driving sequestration into autophagosomes. While several Ub-dependent SARs have been identified and characterized, the E3 Ub ligases that ultimately decide target fate remain poorly studied. In this review, we summarize what is known about the E3 Ub ligases involved in selective macroautophagy, with a particular emphasis on the degradation of mitochondria, protein aggregates, lysosomes and pathogens. A better characterization of these enzymes could improve therapeutic strategies for targeted degradation in acute and chronic diseases.

Alzheimer's disease (AD) and Parkinson's disease (PD) are the two most common age-related neurodegenerative disorders. Allen Human Brain Atlas (AHBA) provides high-resolution transcriptomic data across 102 brain regions with multi-site sampling from healthy controls, promoting the use of brain-wide transcriptomic data for imaging transcriptomics and cross-modal model construction. Increasingly, researchers are utilizing brain-wide transcriptomic datasets to investigate the transcriptome correlates of the neuroimage phenotypes in AD and PD. Leveraging the AHBA, researchers have analyzed the transcriptomic correlations of regional susceptibility to Aβ deposition, tau deposition, α-synuclein propagation, and disease-related multiple-dominal neuroimage phenotypes. These studies revealed that transcriptomic pathways related to metabolism, immunity, neurotransmission, and synaptic function play critical roles in the neuroimage phenotype of AD and PD. By incorporating transcriptomic data modeling, subsequent analyses further confirmed that transcriptomic differences provide the molecular basis for the varying susceptibility observed across brain regions. The analytical approaches of imaging transcriptomics, multimodal data integration strategies, and model construction methods used in AD and PD provide a novel perspective for exploration and can be extended to other neurodegenerative diseases. Future research is expected to utilize brain-wide transcriptomic data to uncover the gene expression mechanisms driving neurodegenerative disease phenotypes.

The mesenchymal homeobox transcription factor MEOX2 is recognized for its roles in development and homeostasis, but the full landscape of its complex and often contradictory functions in human diseases, along with its potential unifying regulatory logic, remains to be systematically elucidated. This Review synthesizes current evidence to provide a comprehensive overview of MEOX2's widespread roles in both neoplastic and non-neoplastic diseases. MEOX2 is remarkably context-dependent: it acts as an oncogene in glioblastoma and lung cancer, but as a tumour suppressor in breast cancer, hepatocellular carcinoma and other tumour types. Its dysregulation is also linked to neurovascular deficits in Alzheimer disease (AD), cardiovascular disorders, metabolic fibrosis and developmental malformations. This functional versatility stems from MEOX2's role as a key signalling integrator, subject to fine-tuned regulation by epigenetic mechanisms, non-coding RNA networks and core pathways including PI3K/AKT, ERK and Hedgehog (Hh). Given the strong association between MEOX2 expression and clinical outcomes, it has emerged as a potential diagnostic and prognostic biomarker for several diseases. Intervention strategies targeting MEOX2 and its regulatory networks show translational promise. This Review proposes a conceptual framework placing MEOX2 as a 'cross-disease core regulatory node', systematically delineates its complex disease associations and molecular mechanisms, and charts a course for the future development of precision medicine strategies targeting MEOX2.

The development of the dentate gyrus (DG) of the hippocampus is protracted over time in comparison with other brain regions such as hippocampal cornus ammoni or neocortex, extending over the first postnatal weeks. During DG postnatal development, neural stem cells (NSCs) will remain to generate the adult neurogenic niche that will sustain granule neuron (GN) production throughout life. NSCs in the DG divide to generate intermediate progenitor cells (IPCs), whose highly regulated dynamics of self-renewal or cell cycle exit decisions still remain poorly understood. Sox5 is a transcription factor (TF) essential for the establishment of adult NSCs, however, its potential role in Sox5 expressing IPCs during DG development remains unexplored. In this study, we demonstrate that conditional loss of Sox5 during embryonic development leads to critical alterations in cell proliferation and survival in IPCs. Specifically, following Sox5 loss, IPCs exhibit a shortening of S-phase duration in late postnatal and juvenile adult stages. Furthermore, these alterations in IPC cell cycle dynamics could be behind the defects in GN differentiation observed in Sox5-defective mice that ultimately leads to subtle morphological changes in DG architecture. Finally, we demonstrate that the additional loss of one Sox6 copy, a closely related TF to Sox5, lead to more profound disruptions in DG morphology than the one observed upon Sox5 loss. Overall, these findings point to a prominent role for Sox5 in combination with Sox6 in IPC cell cycle progression and GN maturation during postnatal DG development.

Ferroptosis is an iron-dependent form of regulated cell death driven by the accumulation of peroxidized phospholipids in cellular membranes. In cancer, susceptibility to ferroptosis is not fixed but instead reflects a dynamic metabolic state shaped by lipid remodeling programs that determine membrane composition, oxidative liability, and the capacity to detoxify lipid peroxides. Tumor cells rewire fatty acid synthesis, desaturation, esterification, storage, sterol metabolism, and ether phospholipid remodeling to alter the abundance and distribution of oxidizable phospholipids and thereby shift their ferroptotic threshold. Polyunsaturated fatty acid-rich membrane phospholipids and di-polyunsaturated phospholipids promote lipid peroxidation and ferroptosis sensitivity, whereas monounsaturated fatty acids, lipid-droplet sequestration, 7-dehydrocholesterol, membrane-bound O-acyltransferase domain-containing 1 and 2, and antioxidant defense systems including glutathione peroxidase 4, ferroptosis suppressor protein 1, and the GTP cyclohydrolase 1-tetrahydrobiopterin pathway suppress ferroptotic death. Therapy-resistant, mesenchymal-like, and drug-tolerant persister states often display elevated oxidative stress together with increased dependence on lipid peroxide detoxification, whereas cancer stem cell-like states can remain either buffered or vulnerable depending on context. Here, we synthesize how lipid-state remodeling, tumor genotype, cell-state plasticity, and microenvironmental cues position tumors along a functional ferroptotic threshold, and we discuss how integrated lipidomic, transcriptional, and state-associated biomarkers may support biomarker-guided ferroptosis-based strategies in precision oncology.

Barrier-to-Autointegration Factor 1 (BANF1) is a self-associating protein encoded within the q13.1 locus of chromosome 11. It is integral to multiple cellular processes, including cell cycle regulation, chromatin organization, gene expression modulation, nuclear envelope (NE) repair, DNA damage repair (DDR), innate immune function, and viral infection control. While prior investigations have provided preliminary phenotypic and bioinformatic characterizations of BANF1 across various disease states, comprehensive analyses detailing its functional roles and mechanistic underpinnings in diverse pathological contexts remain scarce. In cancer, BANF1 is frequently upregulated, with cancer cells leveraging its functions to preserve NE integrity, inhibit activation of the cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) immune pathway, and facilitate epithelial-mesenchymal transition (EMT). These activities contribute to enhanced genomic stability and promote tumor cell proliferation and migration, thereby conferring oncogenic properties. In neurodegenerative diseases, BANF1 is implicated in disease pathogenesis, exemplified by its mediation of glutamate-induced oxidative stress and apoptosis in neuronal cells, as observed in Alzheimer's disease (AD). Additionally, rare mutations in BANF1 are associated with hereditary conditions: the recessive Ala12Thr (A12T) variant causes Néstor-Guillermo progeria syndrome (NGPS), whereas the dominant Gly16Arg (G16R) mutation results in dominant motor neuronopathy. This review further synthesizes recent progress in exploring BANF1 as a therapeutic target, encompassing the development of small-molecule inhibitors, immunomodulatory approaches, and its potential applications in cancer treatment. Overall, this article provides a comprehensive overview of BANF1's structural features, cellular functions, and involvement in disease initiation and progression, with particular emphasis on its expression profiles in malignancies and the therapeutic promise of BANF1-directed interventions.

Ubiquitin-specific protease 25 (USP25), a pivotal deubiquitinating enzyme, has attracted increasing attention in cardiovascular and cerebrovascular research due to its regulatory function in maintaining ubiquitination homeostasis. Considerable advances have been achieved in elucidating the molecular mechanisms through which USP25 participates in the pathological progression of diverse cardiovascular and cerebrovascular disorders. From the perspective of translational medicine, notable progress has also been observed in the development of USP25-targeted therapeutics, with several inhibitors having advanced to preclinical and early-phase clinical trials, thereby offering promising targets for precision treatment. Nevertheless, a systematic synthesis of fundamental biological properties, the trajectory of research development, and the intricate upstream-downstream regulatory networks of USP25, together with its mechanistic roles and therapeutic potential in these diseases, remains absent. In response, this review provides an in-depth summary of the essential biological features of USP25, elucidates the interplay between its upstream regulators and downstream substrates, and highlights its specific molecular mechanisms in cardiovascular and cerebrovascular disease pathogenesis. Additionally, the feasibility and strategic approaches of targeting USP25 for therapeutic purposes are analyzed, while the limitations and potential breakthroughs in the development of USP25 inhibitors are discussed. Finally, priority research directions are proposed to guide future exploration, with the ultimate aim of contributing theoretical foundations and novel perspectives for clinical management of cardiovascular and cerebrovascular diseases.

VISA/MAVS is a central signaling hub that links viral RNA sensing to type I interferon production and inflammatory responses. Because of its potent signal-amplifying capacity, VISA activity must be precisely controlled: insufficient activation compromises antiviral defense, whereas excessive or spontaneous activation can drive chronic inflammation and autoimmune disease. Recent studies have revealed a complex regulatory network governing VISA signaling, involving post-translational modifications, dynamic protein interactions, selective degradation pathways, metabolic cues, and intrinsic inhibitory mechanisms that collectively determine its activation threshold, signaling duration, and downstream output. In this review, we discuss the molecular mechanisms that regulate VISA activation, signal propagation, signal termination, and quiescence maintenance under resting conditions. We propose that immune homeostasis is achieved not through a simple on-off switch, but through continuous regulation of the VISA signalosome life cycle. This framework provides an integrated view of VISA and offers new insights into the molecular mechanisms underlying immune homeostasis.

Primary mitochondrial diseases (PMDs) are one of the most common genetic disorders with an estimated prevalence of 1 in 4300. This review article summarises the latest updates in the field of mitochondrial medicine over the last decade. The availability of exome and genome sequencing in clinical practice has empowered clinicians to unravel the phenotypic heterogeneity of PMD and to end the diagnostic odyssey experienced by many patients and families. In unresolved cases, the detection of variant(s) of unknown significance by next-generation sequencing creates diagnostic and clinical uncertainties, and integrating a multi-omics approach can improve diagnostic yield. Alongside breakthroughs in genomic technologies, there is growing interest in using fluid biomarkers to guide diagnosis, monitor disease progression, and potentially serve as clinical trial endpoints. However, the clinical application of these fluid biomarkers in unselected patient cohorts with different disease onset and phenotypes would require more robust evidence. Natural history studies derived from national and international collaborations have provided insights into genotype-phenotype relationships and prognostic factors across several genotypes, including m.3243A>G, <i>MT-ATP6</i>, <i>POLG</i>, and <i>TK2</i>. Advances in therapeutic discoveries and clinical trials are challenging the obsolete dogma that PMDs are untreatable and bringing hope to patients; four compounds have been licensed, and many trials are in progress. Many barriers and challenges to translating laboratory discoveries into clinical therapy in PMD remain, including preclinical models for efficacy and safety testing, sample size, trial design, and the selection of outcome measures and trial endpoints.

<b>Background/Objectives:</b> Whole exome sequencing (WES) has emerged as a clinically valuable second-tier test following abnormal biochemical newborn screening (NBS). However, population-specific data on diagnostic yield, secondary findings (SFs), and exome-wide carrier burden remain scarce in East Asian neonates, particularly since the release of the ACMG SF v3.3 gene list. We aimed to characterize these metrics in a Taiwanese neonatal cohort. <b>Methods:</b> We retrospectively analyzed 118 consecutive neonates referred to Taipei Veterans General Hospital between August 2021 and August 2022 following abnormal biochemical NBS. WES was performed on the Illumina NovaSeq 6000 platform; variants were classified per the 2015 ACMG/AMP framework and re-evaluated under ACMG SF v3.3. Referral categories comprised lysosomal storage diseases (<i>n</i> = 61), amino acid disorders (<i>n</i> = 38), fatty acid oxidation disorders (<i>n</i> = 13), and organic acid disorders (<i>n</i> = 6). <b>Results:</b> Fifty neonates (42.4%) received confirmed molecular diagnoses and 45 (38.1%) were carriers (combined molecular resolution 80.5%). Five participants (4.2%) harbored pathogenic/likely pathogenic variants in ACMG SF v3.3 genes (<i>TTN</i>, <i>LDLR</i>, <i>PTEN</i>, <i>RYR1</i>, <i>TP53</i>). Incidental findings occurred in 51.7%, and at least one recessive-carrier variant was detected in 99.2% (mean 4.15 per individual; median 4). The Taiwanese-specific c.639+919G>A cardiac Fabry variant accounted for 24/25 confirmed male Fabry cases, and the p.Gly576Ser pseudodeficiency allele confounded all suspected Pompe cases. <b>Conclusions:</b> Second-tier WES substantially improves diagnostic precision, discriminating confirmed diagnoses from carrier, pseudodeficiency, and biochemical false-positive states. The high carrier burden and incidental-finding rate underscore the importance of comprehensive pre- and post-test genetic counseling in East Asian neonatal genomic programs.

The most common symptom of endometriosis is pain. The pathogenesis of endometriosis-associated pain remains unclear. In this review, we illustrate the pathogenesis of endometriosis-associated pain and the mechanisms of dienogest acts on endometriosis-associated pain, as well as therapeutic applications targeting these processes, by searching the literature in PubMed and Web of Science until May 2026. Abnormal distribution of nerve fibers, central sensitization, imbalance of inflammatory and immune environment, and hypoxic stress contribute to the generation of endometriosis-associated pain. Dienogest, as a fourth-generation selective progesterone receptor agonist, plays a key role in the treatment of endometriosis-associated pain by reducing serum estrogen levels, alleviating inflammatory responses, regulating nerve growth factor, modulating control central nervous system allergies, and influencing the biological processes of endometriotic stromal cells. Moreover, its long-term safety and tolerability are relatively satisfactory according to clinical studies, making it an effective therapeutic agent for endometriosis.

<b>Background:</b> Glutamatergic neuron-to-glioma signaling mediated by α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors (AMPARs) has emerged as an important mechanism in glioma progression. <b>Objectives/Methods:</b> We analyzed the expression of the AMPAR subunit genes <i>GRIA1</i>, <i>GRIA2</i>, <i>GRIA3</i>, and <i>GRIA4</i> in lower-grade glioma (LGG). <b>Results:</b> Expression of <i>GRIA1</i>-<i>GRIA4</i> was highest in IDH-mutant/1p19q-codeleted tumors and lowest in IDH-wildtype tumors across both The Cancer Genome Atlas (TCGA) and the Chinese Glioma Genome Atlas (CGGA) cohorts. High expression of each <i>GRIA</i> gene was associated with longer overall survival (OS). Transcriptome-wide analyses identified positive correlations between an AMPAR score and genes involved in synaptic organization, neuronal connectivity, and neurotransmission. Co-expression analyses demonstrated coordinated expression between <i>GRIA1</i>-<i>GRIA4</i> and genes encoding AMPAR auxiliary proteins. Gene Ontology (GO) enrichment revealed overrepresentation of synaptic signaling, trans-synaptic communication, and synapse organization. Although the AMPAR score was associated with favorable survival in univariate analyses, it did not retain independent prognostic significance after adjustment for key clinicomolecular variables. Elevated expression of AMPAR subunit genes in LGG was associated with favorable molecular subtypes and a synaptic transcriptional program. <b>Conclusions:</b> These findings suggest that <i>GRIA1</i>-<i>GRIA4</i> expression is associated with a synaptically enriched transcriptional program in LGG, although its cellular origin remains uncertain.

<title>Abstract</title> <p>Background Digital cognitive testing and plasma biomarkers have emerged as key players for population screening and measuring drug efficacy in clinical trials for Alzheimer’s Disease (AD). Understanding test-retest reliability of these tools is essential to estimate meaningful effects of an intervention over time. Whilst some data exist on variability of plasma and digital biomarkers at very short time frames or annually, studies conducted at intermediate time scales are lacking. Data on impact on this variability on clinical decision-making are also rare. Methods Paired testing of plasma biomarkers and digital cognition was performed at 0, 3 and 6 months in a cohort of 92 individuals (55 cognitively unimpaired healthy controls (HC) and 37 patients with early AD dementia (AD)). Plasma biomarkers examined included pTau217, pTau181, NfL, GFAP and the Aβ42/40 ratio. OCTAL (Oxford Cognitive Testing Portal) digital cognitive testing platform was used to measure cognition. Results pTau181 emerged as the plasma biomarker with worst test-retest reliability and was the only measure where repeating the sample improved accuracy in group discrimination. None of the digital measures showed further improvement in diagnostic accuracy if repeated over time, reflecting high reliability. Individual metrics showed different reliability profiles based on the index used to measure variability (Coefficient of variation or Intraclass Correlation Coefficient) and varied within groups (HC and AD). Conclusion Test-retest reliability measures are essential for interpretation of meaningful effects over time for plasma and digital outcomes.</p>

<b>Aim:</b> Metabolic dysfunction-associated steatohepatitis (MASH) is a progressive form of metabolic dysfunction-associated steatotic liver disease, linked to hepatic and extra-hepatic complications and substantial healthcare costs. Despite its clinical and economic impact, real-world evidence on disease progression and costs in Germany is limited. <b>Materials & methods:</b> We conducted a retrospective cohort study using statutory health insurance claims from the InGef database (2016-2023), covering 4.7% of the German population. Patients with MASH were identified using ICD-10-GM code K75.8, in absence of the more specific code in the German coding system. Baseline characteristics and comorbidities were assessed over 2 years prior to index diagnosis. Progression was defined by transitions through end-stage liver disease (ESLD) stages: compensated cirrhosis, decompensated cirrhosis, hepatocellular carcinoma and liver transplantation. Healthcare costs were analyzed descriptively and via regression models. <b>Results:</b> Among 4710 patients with MASH (prevalence: 0.15%), 39.4% had documented ESLD during follow-up (mean follow-up in days 1530). Disease progression to ESLD occurred in 26.0% of patients without baseline ESLD (n = 922/3490), whereas 4.0% of patients with baseline ESLD (n = 48/1188) progressed to a more severe ESLD stage during follow-up, with a mean time to first progression of approximately 25.3 months. Patients with ESLD incurred annual costs of €12,737 versus €4928 for those without ESLD. Progression to hepatocellular carcinoma and liver transplantation resulted in predicted costs of €12,948 and €75,719 per patient-year, respectively. Mortality was significantly higher among patients with ESLD (incidence rate ratio: 4.65; 95% CI: 3.76-5.79). <b>Discussion:</b> Over a quarter of identified patients with MASH already had advanced liver disease at baseline, suggesting potential underdiagnosis or late recognition in routine clinical practice. Early detection, proactive management and targeted therapies are essential to reduce progression and economic burden.

Dual-specificity protein kinase CLK4 plays a pivotal role in regulating alternative mRNA splicing, DNA repair, and various cellular processes through precise phosphorylation. In this study, we analyzed 3825 global human cellular phosphoproteome studies, identifying 430 qualitative profiles and 55 quantitative differential datasets featuring high-confidence Class-1 phosphosites (localization probability ≥ 75%; A-score ≥ 13). Notably, S136 and S138 emerged as predominant phosphorylation sites outside the kinase domain. These sites showed frequent detection and differential expression in liver, lung, and head and neck cancers, as documented in PhosphositePlus. We identified a high-confidence set of co-regulated phosphoproteins, including SQSTM1, SRRT, RPS6, TP53BP1, TNKS1BP1, THUMPD1, OTUD4, and TCEA1. These proteins link CLK4 to critical pathways, including RNA splicing, autophagy, DNA damage response, and cancer progression. Binary interactors, including SRRM2, Interacts with SPT6 1 (IWS1), RBBP6, ZC3H18, BUD13, and DYRK1A, further connect CLK4 to RNA processing and splicing. Predicted downstream substrates, such as CCNL2, CDK11B, PRDX6, YTHDC1, RBM15, SRRM1, SFSWAP, HNRNPU, and DOCK7, highlight CLK4's broad regulatory scope. Upstream kinases were also predicted for S136 and S138. Site-resolved analyses revealed tumor-specific dysregulation of CLK4 phosphorylation at key residues. Co-occurring phosphosite alterations and nearby somatic mutations suggest disrupted CLK4 regulation in cancer. Overall, this study provides a comprehensive phosphoproteomic resource that maps CLK4's co-regulatory networks, paving the way for mechanistic investigations and targeted cancer therapies.

Mitochondrial gene expression is essential for oxidative phosphorylation that generates the bulk of the cellular ATP, and mitochondrial dysfunction is a common cause of human metabolic diseases. Recently, the first pathogenic variants in the only known mitochondrial RNA polymerase (POLRMT) were described in patients presenting with a wide variety of clinical manifestations, including hypotonia, short stature, and developmental delay. Here, we modeled two human pathogenic POLRMT variants by creating the corresponding substitutions in mice: the dominant S582F and the recessive R984C variant. Mice homozygous for the R984C variant showed perinatal lethality without apparent embryonic developmental defects, a finding consistent with a failure to adapt to the metabolic transition to oxidative metabolism at birth. Mice carrying the S582F variant were viable and exhibited decreased mitochondrial transcript levels due to impaired de novo transcription. However, mtDNA levels and in organello mtDNA replication remained normal, which recapitulates the molecular phenotypes observed in patients. Altogether, our findings indicate that the conserved arginine near the active site is essential for POLRMT function, while the serine in the intercalating hairpin of the N-terminal domain is required for near-genome length transcription but not primase activity. This study highlights genotype-phenotype differences and provides new insights into POLRMT function.

<h4>Background</h4>Centrosome-related genes (CRGs) regulate cell division and genomic stability and may influence tumor progression, but their prognostic and functional roles in bladder cancer (BLCA) are not fully defined.<h4>Methods</h4>We curated 698 CRGs and identified 12 prognostic genes to construct a centrosome-related signature score (CRSS). The prognostic value of CRSS was evaluated in TCGA and GSE13507 cohorts. Functional enrichment, immune landscape, and drug sensitivity were analyzed through pathway analysis, ESTIMATE, and IC50 prediction. Machine learning and Mendelian randomization were used to identify the most promising CRGs. Furthermore, we performed single-cell RNA sequencing analysis to uncover the expression patterns of candidate CRGs. Expression and functional validation were performed using RT-qPCR, CCK-8, wound-healing assays, and immunohistochemistry.<h4>Results</h4>CRSS stratified patients into high- and low-risk groups with significant differences in overall survival. High-risk patients exhibited upregulated Aerobic glycolysis (AC), oxidative phosphorylation (OxPhos) and partial epithelial-mesenchymal transition (pEMT) pathways, higher immune/stromal scores, and distinct immune infiltration, including increased M0/M2 macrophages and decreased CD8⁺ T and follicular helper T cells. High-risk tumors showed altered sensitivity to multiple drugs, including AZD8186 and Staurosporine. Integrative analyses identified SPHK1 as a key candidate CRG. SPHK1 expression was associated with tumor grade, stage, poor survival, and the wound-healing molecular subtype. Functional assays showed that SPHK1 knockdown inhibited bladder cancer cell proliferation and migration. Immunohistochemistry further confirmed higher SPHK1 expression in tumors with more aggressive clinicopathological features.<h4>Conclusions</h4>We established a CRG-based model for BLCA and identified SPHK1 as a key gene associated with tumor progression and clinical outcomes.

The population of Vietnam remains underrepresented in global genomic databases. Here, we present VN1K, a resource of multi-omics and phenotypic information for 1011 unrelated Vietnamese individuals. We present high-depth short-read whole-genome sequencing data for all samples along with various -omics datasets. Using a high-sensitivity variant detection pipeline, which includes a pangenome graph reference and a deep-learning framework, we identify approximately 42 million variants with 7 million short insertions/deletions and 90 thousand structural variants. VN1K also features a whole-genome methylation profile based on long read sequencing. We create a genotype imputation panel with high accuracy on the Vietnamese population, allowing us to identify variants with significantly different allele frequencies in the Vietnamese population compared to other populations. We establish the functional relevance of some of these variants, particularly those in genes associated with genetic disorders, immune diseases, and drug responses, by integrating the allele frequency differences with known genotype-phenotype associations and clinical annotations. Further, we map various loci related to hepatitis B virus infection, triglyceride levels, LDL-C levels, serum glucose levels, HbA1c levels, and levels of two liver enzymes (ALT and AST). The VN1K dataset is accessible via genome.vinbigdata.org, an integrated platform with both linear and graph-based genome browsers.

The nuclear genome is spatially organized into a three-dimensional architecture by physical association of large chromosomal domains with subnuclear compartments including the nuclear lamina at the radial periphery and nuclear speckles within the nucleoplasm^1-5. However, how higher-order spatial genome architecture regulates human development has been overlooked, and the interplay between chromatin state and subnuclear genome compartmentalization is poorly understood. Here we generate high-resolution maps of genomic interactions with the lamina and speckles in cells of the neurogenic lineage isolated from mid-gestational human cortex, identifying an intimate association between subnuclear genome compartmentalization, chromatin state and transcription. During cortical neurogenesis, subnuclear genome compartmentalization is extensively remodelled, relocating hundreds of neuronal genes from the lamina to speckles, including key neurodevelopmental genes bivalent for trimethylation of histone H3 at Lys27 (H3K27me3) and Lys4 (H3K4me3). At the lamina, bivalent genes have exceptionally low expression, and relocation to speckles enhances resolution of bivalent chromatin to H3K4me3 monovalency and increases transcription more than eightfold. We further demonstrate that proximity to the nuclear periphery-not the presence of H3K27me3-maintains the lowly expressed, poised state of bivalent genes embedded in the lamina. We find that the repressive environment of the lamina is associated with spatial segregation of the transcriptional elongation machinery from the nuclear periphery. Our results establish a paradigm in which knowing the spatial location of a gene is necessary for understanding its epigenomic regulation.

Accurate single-molecule optical genome mapping remains challenging due to the limited information content of barcode-only or profile-only strategies. Here, we present Dual Optical Mapping (DOM), which integrates sequence-specific barcode markers with AT frequency-dependent intensity profiles on the same DNA molecules to enhance mapping accuracy. Conventional optical mapping approaches rely either on sequence-specific barcodes generated by restriction endonucleases, nicking enzymes, or methyltransferases, or on dense profile mapping using DNA-binding molecules. By combining these complementary sources of information within a unified dual-channel framework, DOM increases positional specificity and alignment confidence. As a model system, we applied DOM to the E. coli genome (4.6 Mbp), where the combined dual-channel scoring metric (cc_rg2) ranked 172 of 182 molecules (95%) at the correct genomic locus as the top match, while 179 of 182 molecules (98%) could be validated after quality-control review. We further extended DOM to the human genome through genome-wide cross-correlation scanning coupled with placement score ranking. This quantitative framework enables objective evaluation of alignment distinctiveness across the entire genome, demonstrating scalability to large and complex genomic contexts. These results establish DOM as a scalable dual-channel framework for high-accuracy single-molecule genome mapping across both bacterial and human genomes.

Classical Major Histocompatibility Complex Class I (MHC-I) molecules are traditionally viewed as stable peptide-presenting structures expressed on the surface of all nucleated cells. Their expression by professional antigen-presenting dendritic cells (DCs) enables CD8+ T-cell activation, differentiation, and immune surveillance. However, accumulating evidence indicates that cell-surface MHC-I molecules exist in three major conformational states: (1) β2m-associated, peptide-loaded conformers that originate in the endoplasmic reticulum and pass through the Golgi apparatus after binding proteasome-generated cytosolic peptides (hereafter referred to as closed conformers); (2) β2m-free, peptide-empty conformers that arise following β2m dissociation from closed conformers either at the plasma membrane or after internalization and recycling (hereafter referred to as open conformers); and (3) β2m-associated, peptide-empty conformers that represent an intermediate state between closed and open conformers. Here, we propose a conceptual framework, supported by computational predictors of intrinsically disordered regions, in which transitions between closed and open MHC-I conformers are coupled to intracellular regulatory processes, including post-translational modifications of conserved motifs, intracellular trafficking, and signaling. Although direct experimental evidence linking these processes remains limited, we integrate independent observations into a working model that may guide future investigations into MHC-I-mediated cell-cell communication in both immune and non-immune contexts, in health and disease. For clarity, in this article we define "open conformers" as structurally competent, β2m-free, and peptide-deficient MHC-I molecules.

Salivary adenoid cystic carcinoma (SACC) is an immunologically cold malignancy with limited response to current immunotherapies. Integrated transcriptomic profiling of peripheral blood, primary tumors, lung metastases, and a two-donor single-cell dataset revealed compartment-specific expression signatures and, through computational inference, systemic immune dysregulation marked by hematopoietic suppression, T-cell exhaustion, compensatory myelopoiesis, and an immature B-cell expansion. To explore the transcriptional basis of this peripheral immune aberration, blood-derived RNA-seq was interrogated, identifying only 32 unique genes meeting |log2FC| > 1 and <i>q</i> < 0.05 among 34,999 transcripts; qPCR confirmed concordant upregulation of <i>IL33</i> and <i>CCL14</i>, providing directional rather than confirmatory support, suggesting peripheral immune molecular aberrations that still require validation through broader differential gene expression validation. Complementing this transcriptomic signature, detection of <i>MYB-NFIB</i> fusion transcripts matching tumor tissue in one patient's blood suggested that tumor-derived signals may access the circulation, although cohort validation remains necessary. Extending these peripheral observations to tissue compartments, we applied expression stratification, correlation networks, ligand-receptor mapping, and a virtual gain-loss model to computationally predict regulatory associations involving <i>IL17RB/OLIG1/NOTCH1</i> in primary tumors and a <i>CD24/IL17RB/MYB/MYBL2/CXCL13/CXCR5</i> module in lung metastases. At the single-cell level, cluster 10 emerged as a cell-cycle-high tumor population with transcriptional overlap with proliferating immune progenitors, providing a potential cellular basis for tumor cell entry into the circulation. Collectively, these computational inferences generate testable hypotheses for multicompartment immune dysregulation in SACC, positioning <i>IL17RB</i> as a candidate molecule that warrants prospective validation in SACC-specific preclinical models.

<h4>Introduction</h4>Sickle cell disease is a monogenic blood disorder prevalent in Africa, where malaria poses a health risk for affected individuals. Individuals with sickle cell disease are susceptible to malaria, which can be complicated by vaso-occlusive episodes, severe anaemia and death. Understanding the intersection of malaria and sickle cell disease in African countries with high co-prevalence is essential for developing effective management and prevention strategies, including clinical guidelines on the use of malaria chemoprophylaxis in this population. This study will determine the monthly incidence of malaria in children living with sickle cell disease in each of two clusters of African countries.<h4>Methods and analysis</h4>This 1-year observational study will enrol 2808 children with sickle cell disease, aged 6 months to 17 years, at Sickle Pan-African Research Consortium clinical sites in Ghana, Mali, Nigeria (West), Tanzania, Uganda, Zimbabwe and Zambia (East-South). Malaria infection will be assessed at monthly visits and unscheduled sick visits over 12 months. Data collected will include socio-demographic characteristics, beta-globin genotype, medical history, clinical symptoms and malaria blood smear or rapid diagnostic test results. The primary outcome is monthly incidence of clinical malaria; a secondary outcome is incidence of asymptomatic malaria.<h4>Ethics and dissemination</h4>This study was developed with input from patients with sickle cell disease and their caregivers. The study will be conducted according to the principles of the Declaration of Helsinki. Written informed consent from guardians and assent from child participants age 7 or above will be provided before enrolling in the study. This protocol has been reviewed and approved by local and national ethics committees at each study site. The findings of this study will be provided to participants and the general public through presentations at the annual Sickle Cell Day community events at each study site, local media and open access publication in peer-reviewed journals.The full names of the ethics committees are as follows. Ghana: Komfo Anokye Teaching Hospital Institutional Review Board; Mali: Ethics Committee of the University of Sciences, Techniques and Technologies of Bamako; Nigeria: National Health Research Ethics Committee of Nigeria; Tanzania: Muhimbili University of Health and Allied Sciences Research Ethics Committee and National Health Research Ethics Committee; Uganda: Makerere School of Medicine Research and Ethic Committee; Zambia: National Health Research Authority and Excellence in Research Ethics and Science Converge Institutional Review Board; and Zimbabwe: Joint Research Ethics Committee and the Medical Research Council of Zimbabwe.

Langer mesomelic dysplasia is an exceptionally rare skeletal dysplasia caused by complete or functionally complete deficiency of the <i>SHOX</i> (short stature homeobox) gene located within the pseudoautosomal region 1 (PAR1) of the sex chromosomes. Clinically, the disorder is characterized by severe disproportionate short stature and marked mesomelic shortening of the limbs, particularly involving hypoplasia or aplasia of the ulna and fibula, while cognitive development and life expectancy are generally preserved. This narrative review summarizes current knowledge regarding the molecular genetics, developmental biology, clinical manifestations, radiographic findings, prenatal diagnosis, and differential diagnosis of Langer mesomelic dysplasia. The SHOX protein functions as a homeodomain-containing transcription factor essential for chondrocyte proliferation, differentiation, and growth plate organization. Pathogenic mechanisms include biallelic <i>SHOX</i> deletions, enhancer-region defects, missense variants affecting the homeodomain and nuclear localization signal, as well as splice-site variants leading to severe reduction of functional protein dosage. The article also discusses the broad phenotypic spectrum of <i>SHOX</i> deficiency, genotype-phenotype variability, and the relationship between Langer mesomelic dysplasia and related disorders such as Léri-Weill dyschondrosteosis and Turner syndrome. Understanding the molecular basis of this condition is essential for accurate diagnosis, genetic counseling, and prenatal assessment in affected families.

<h4>Background</h4>Sepsis and acute kidney injury (AKI) are life-threatening conditions often coexisting as sepsis-associated AKI (S-AKI). However, their shared molecular mechanisms and immune heterogeneity remain unclear. This study aims to identify robust diagnostic biomarkers applicable to both conditions and to elucidate their diverse immune microenvironments using integrated transcriptomic approaches.<h4>Methods</h4>Transcriptomic datasets for sepsis and AKI were analyzed, with multiple cohorts used for training and external validation. Differential gene expression and WGCNA identified key modules, while LASSO and Random Forest algorithms screened shared hub genes. A diagnostic nomogram was constructed and evaluated using ROC and decision curve analyses. Single-cell RNA sequencing data were further analyzed to determine cellular localization, functional pathways, and intercellular communication.<h4>Results</h4>Four hub genes (<i>FBXO21</i>, <i>FLOT1</i>, <i>TMC6</i>, and <i>KLRB1</i>) were identified as robust diagnostic biomarkers for both sepsis and AKI, demonstrating strong predictive performance across validation cohorts. Single-cell analysis revealed that these genes were enriched in specific immune cell populations and injured renal cells, and were closely associated with T-cell activation and immune signaling pathways. Cell-cell communication analysis further inferred distinct ligand-receptor interactions that may underlie immune crosstalk in both conditions.<h4>Conclusion</h4>We identified a reliable four-gene diagnostic signature shared by sepsis and AKI and characterized their shared immune heterogeneity and inferred intercellular communication networks. These findings provide potential targets for early diagnosis and therapeutic intervention in sepsis-associated renal injury.

<h4>Introduction</h4>Duolang sheep is a meat-fat dual-purpose breed originating from Xinjiang, China, and the genetic basis of body size variation in this breed remains unclear.<h4>Methods</h4>Whole-genome resequencing was performed on 224 Duolang sheep with complete records for six body size traits. After variant calling, imputation, and quality control, 19.65 million high-quality SNPs were retained. After linkage disequilibrium pruning, 2.1 million relatively independent SNPs were used for a mixed linear model GWAS in GEMMA. Tajima's D and integrated haplotype score analyses were performed in a kinship-filtered subset of 60 animals, and SheepGTEx data were used to characterize tissue expression patterns.<h4>Results</h4>The GWAS identified 24 significant SNPs for height, body length, chest girth, shoulder width, and hip width. Integration of GWAS intervals with selection-signal regions prioritized 31 candidate genes, including <i>CA10</i>, <i>WDR45B</i>, <i>FADS2</i>, <i>LGALS12</i>, and <i>CTNNA2</i>. Multi-tissue expression profiles provided functional context for the prioritized genes.<h4>Discussion</h4>These findings provide insight into the genetic architecture of body size traits in Duolang sheep and identify candidate loci for future validation and genetic improvement studies.

Cancer remains a major global health burden, with its incidence and mortality rates persistently high despite advances in treatment. Despite therapeutic innovations, malignant tumors continue to pose a formidable challenge to global health. Against this backdrop, the crosstalk between long non-coding RNAs (lncRNAs) and the Notch signaling pathway has emerged as a pivotal driver of tumorigenesis and progression. However, the complex regulatory network and a comprehensive mechanistic framework of this axis await systematic elucidation. This review systematically consolidates recent advances in understanding how lncRNAs precisely modulate Notch pathway activity through diverse mechanisms, including acting as competing endogenous RNAs, direct protein binding, epigenetic regulation, and exosome-mediated intercellular communication. The discussion encompasses various malignancies, spanning the digestive, respiratory, urogenital, nervous, and hematologic systems. The lncRNA-Notch regulatory axis is identified as a ubiquitous and functionally central oncogenic network. It orchestrates critical malignant phenotypes-such as such as stemness maintenance, epithelial-mesenchymal transition, metabolic shifts, drug resistance, and immune evasion-through intricate bidirectional crosstalk. Functional studies confirm that targeting key nodes of this axis can effectively reverse drug resistance and suppress tumor growth. Although challenges remain in its clinical translation, future research integrating single-cell multi-omics, nanotechnology, and other innovative strategies will undoubtedly open new avenues for precision diagnosis and cancer therapy.

<h4>Introduction</h4>Lactylation, an emerging post-translational modification, modulates tumor metabolism and gene expression, thereby influencing the initiation and progression of cervical squamous cell carcinoma (CESC). This study aims to identify lactylation-associated prognostic features in CESC through the integration of transcriptomic data and single-cell RNA sequencing (scRNA-seq).<h4>Methods</h4>Publicly available datasets were utilized for this study. Lactylation-related genes (LRGs) linked to CESC prognosis were found through analysis. Key prognostic genes include PGK1, PFKP, DDX39A, RFC4, WAS, and PFKM, with PFKP identified as a risk factor.<h4>Results</h4>The risk model demonstrated strong predictive performance, and immune infiltration analysis revealed elevated levels of immune cells, with CD8⁺ T cells negatively correlating with risk. ScRNA-seq analysis identified three distinct cell types, with CD8⁺ T cells showing a distinct developmental trajectory, marked by a reduction in early-stage cells and an accumulation of late-stage cells within the CESC microenvironment. Moreover, the expression of prognostic genes was elevated during the later stages of CD8⁺ T cell differentiation.<h4>Conclusion</h4>LRGs demonstrated significant prognostic value in CESC and accurately predicted patient outcomes. Furthermore, the study underscored the pivotal role of CD8⁺ T cells in the progression of CESC.

<title>Abstract</title> <p>Exosomes play critical roles in prostate cancer (PRAD) progression. This study aimed to identify and validate exosome-related diagnostic biomarkers for PRAD using integrated bioinformatics and machine learning approaches. Transcriptomic data from TCGA-PRAD and three GEO cohorts were intersected with curated exosome-related gene sets to obtain exosome-related DEGs (Exo-DEGs). Core diagnostic genes were selected through a consensus strategy integrating LASSO, SVM-RFE, and random forest. Diagnostic performance was assessed using ROC analysis, nomogram, and decision curve analysis, while model interpretability was examined using SHAP. Immune infiltration was estimated by ssGSEA. Drug prediction used DSigDB enrichment and molecular docking to AKR1B1. Twelve Exo-DEGs were identified, yielding an eight-gene exosome-related diagnostic signature (CAV1, CLU, FGFR2, APOE, AKR1B1, OLFM4, ALB, and GATA4). The signature demonstrated excellent diagnostic accuracy (AUC = 0.966), outperforming single-gene classifiers. Among ten machine-learning classifiers, random forest achieved the best test-set performance (AUC = 0.953), with SHAP highlighting FGFR2, APOE, CAV1, and CLU as top contributors. Core genes were positively associated with multiple immune cell signatures. Molecular docking suggested stable AKR1B1 binding with cholesterol (Vina score − 8.2 kcal/mol) and warfarin (− 8.0 kcal/mol). Bulk analyses showed decreased AKR1B1 expression in tumors, while single-cell analyses revealed prominent expression in myeloid populations, particularly macrophages, linked to inflammasome signaling, EMT, and immune responses. AKR1B1 represents a promising diagnostic biomarker with functional relevance to tumor immunity and metabolism, offering potential for targeted drug development.</p>

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease characterized by the selective degeneration of motor neurons with a substantial genetic contribution to disease risk. Distinct genetic etiologies converge on key pathogenic processes in ALS, including aberrant RNA metabolism, proteostatic stress, dysregulated vesicular transport, and mitochondrial dysfunction. Here, we investigate whether perturbed transcriptional programs associated with definitive ALS genes harbor elusive genetic contributors to disease. We analyzed RNA sequencing data profiling induced pluripotent stem cell-derived motor neurons from individuals with ALS ( N = 438) and controls ( N = 187). Leveraging an integrative analytical framework, we identified transcriptional pathways linking variants in definitive ALS genes to widespread coordinated co-expression activity driven by gene-specific transcriptional programs. We subsequently subjected all genes comprising significant pathways to genetic analyses to uncover novel targets and characterized these genes using single-nuclei transcriptomics of the motor and prefrontal cortices of individuals with ALS and controls. The identified transcriptional programs associated with definitive ALS genes showed strong concordance to established biology, including dysregulated proteostasis and mitochondrial stress in SOD1 ALS, protein aggregation and degradation pathways in FUS ALS, and altered vesicle dynamics, synaptic transmission, autophagy, and trafficking programs in GRN ALS. These pathways encompassed both well-established ALS genes, as well as six target genes— PTPRN2 , UNC13C , TTC3 , USP10 , PSMD4 , and RUFY3 —of which USP10 , PSMD4, and RUFY3 have not yet been described in the context of ALS genetic association studies. Single-nuclei analyses demonstrated that target gene expression was enriched in the baseline architecture of neuronal populations, supporting neuron-intrinsic mechanisms of vulnerability. PSMD4 showed region-specific dysregulation in selectively vulnerable Layer 5 motor neurons of individuals with ALS, implicating regionally divergent proteasome activation as a neuron-intrinsic feature of ALS vulnerability. Collectively, this work advances the molecular understanding of distinct genetic ALS etiologies, provides mechanistic insight into emerging ALS target genes, and establishes a generalizable framework that is readily applicable across heritable diseases for the identification of disease-relevant molecular pathways and target genes.

Induced pluripotent stem cells (iPSCs) are prone to genomic instability during prolonged culture, with recurrent chromosomal aberrations conferring selective advantages. Replication stress is a major driver of this instability, yet the repertoire of replication stress‑sensitive loci in iPSCs remains largely unexplored. Here, we mapped aphidico-lin‑sensitive fragile sites (asFS) in three independent iPSC lines using classical cytogenetic break analysis combined with Monte Carlo simulation and MiDAS mapping directly on banded metaphase chromosomes. We identified 28 asFS, which segregated into a highly active Major cluster (8 sites, accounting for 59% of breaks among asFS) and a less active Minor cluster (20 sites). Five universal asFS (9p21, 6q25-26, 20p11-12, 10q22, Xq25) were present in all three lines, representing a tissue‑specific fragility signature of pluripotent stem cells, with Xq25 shifting into the Major cluster after correction for X chromosome dosage. Minor asFS showed preferential co‑localization with physical breakpoints or minimal overlapping regions of recurrent culture‑acquired aberrations, including 20q11.21 (BCL2L1), 1q32 (MDM4), 8q24 (MYC), 17q21 (WNT3‐WNT9B), and 18q21 (DCC/FRA18B). MiDAS mapping validated most asFS and revealed additional replication stress‑sensitive loci in pericentromeric and subtelomeric regions that are difficult to score by conventional G‑banding. Comparison with fragile site maps from other cell types con-firmed tissue specificity, showing that the iPSC asFS repertoire is distinct in rank order and relative activity. Collectively, our findings indicate that the asFS repertoire in iPSCs is hierarchically organized into a stable universal core and a variable peripheral component, and suggest that Minor asFS may serve as initial substrates for adaptive culture‑acquired rearrangements. This work provides a framework for understanding how replication stress and clonal selection shape the mutational landscape of pluripotent stem cells

Huntington’s disease (HD) is a progressive neurodegenerative disorder caused by an expanded CAG repeat in the huntingtin ( HTT ) gene. The disease is characterized by movement disorders, and it also presents with personality changes, including apathy and aggression, along with cognitive decline. While most animal models for HD have been validated for motor deficits, less is known about alterations in other behavioral functions. Here, we performed a longitudinal study to analyze the behavior of a knock-in mouse model of HD with a chimeric mouse/human exon 1 containing 140 CAG repeats inserted in the murine huntingtin gene. We specifically inquired about the onset of cognitive impairments in knock-in mice and whether changes in various behavioral functions such as locomotion, anxiety, and cognition correlate at the individual level. Our data indicate that female and male knock-in mice exhibit reductions in body weight, novelty-induced locomotion, and remote spatial memory retrieval. However, social behavior, working, and short-term memory remain unaffected. Within knock-in mice, lower open-field activity correlated with poorer remote memory performance. Moreover, CAG repeat length negatively correlated with locomotor activity and spatial memory, indicating that greater repeat expansion predicts more severe behavioral impairment. These findings identify early affective changes, followed by selective long-term memory and locomotor deficits, in knock-in mice, supporting this model as a useful platform for studying prodromal HD and repeat-length-dependent disease variability. <h4>Highlights</h4> CAG140 knock-in mice show early anxiety, later reduced locomotion and memory deficits Long-term and remote memory are impaired while short-term and working memory are spared Lower locomotion at the age of 8 months correlates with poorer memory at 14 months of age in individual CAG140 knock-in mice Greater CAG repeat length predicts worse locomotion and memory

Endometrial injury remains a leading cause of female infertility, yet current clinical interventions exhibit limited regenerative efficacy. In this study, we developed a micro-nano composite system (G-H/ADSC@PDA/PFD) combining adipose-derived mesenchymal stem cell (ADSC)-laden double-network hydrogel (G-H/ADSC) microspheres with pirfenidone (PFD)-loaded polydopamine (PDA/PFD) nanoparticles to achieve synergistic stem cell and drug delivery. The microspheres were fabricated from gelatin methacryloyl (GelMA) and hyaluronic acid methacryloyl (HAMA) via droplet microfluidic technology, exhibiting favorable mechanical strength and cytocompatibility. They provide a 3D microenvironment that supports ADSC growth, proliferation, and paracrine secretion, thereby enhancing angiogenesis and modulating the inflammatory milieu. Moreover, the incorporated PDA/PFD nanoparticles enhance the adhesion of the composite system to the injured endometrial site, while efficiently scavenging reactive oxygen species. The sustained release of PFD downregulates TGF-β expression, inhibits collagen deposition and fibrotic progression, and further optimizes the regenerative niche for ADSCs. In a rat model of endometrial injury, the G-H/ADSC@PDA/PFD system significantly promoted endometrial regeneration, reduced fibrosis, restored the local microenvironment, and successfully restored fertility. This study introduces a promising combinatorial strategy for the treatment of endometrial injury and infertility.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by amyloid-β (Aβ) deposition. The liver-brain axis underscores the central role of the liver in modulating cognitive function through multidimensional regulatory mechanisms. As a core metabolic and detoxification organ, the liver also helps maintain cerebral homeostasis via pathways including the urea cycle, antioxidant systems, ketone body metabolism, and bile acid regulation. Dysfunction of these processes may lead to ammonia accumulation, exacerbated oxidative stress, and Aβ clearance, thereby accelerating the pathological progression of AD. Liver-derived factors such as apolipoprotein E (APOE), C-reactive protein (CRP), fibroblast growth factor 21 (FGF21), and insulin-like growth factor 1 (IGF-1) significantly increase the risk of AD through dual mechanisms-inhibiting Aβ clearance and activating neuroinflammation, thereby directly affecting cognitive function via modulation of inflammation, metabolism, and blood-brain barrier (BBB) integrity. Neural interfaces formed by the hypothalamic-pituitary-target gland axis and the vagus nerve enable communication from the liver to the brain, with emerging evidence also supporting a reverse influence from the brain to the liver. Emerging technologies such as molecular tracing and nanocarriers provide new tools for deciphering dynamic interactions within the liver-brain axis. Liver-targeted metabolic interventions show potential for reversing cognitive impairment. Unlike previous reviews that mainly focused on single pathways, this review conceptualizes the liver-brain axis as a multidimensional regulatory network in AD. By clearly linking network nodes to potential therapeutic interventions, it provides us with a novel framework that not only describes the various mechanisms but also focuses on identifying actionable targets for disease prevention and treatment.

Germline variants in leukemia predisposition genes are increasingly detected in pediatric patients. Nevertheless, the rare variants identified in diagnostic samples may be misinterpreted without variant-level functional evaluation and cautious clinical interpretation. Whole-exome sequencing was performed on 28 individuals from five kindreds in which at least two children developed acute leukemia. Results identified a rare ETV6 variant, c.604C > G (p.Arg202Gly), in monozygotic twins with ETV6::RUNX1-positive B-cell precursor acute lymphoblastic leukemia. To support variant interpretation, HeLa-cell populations stably expressing FLAG-tagged wild-type ETV6 and p.Arg202Gly and the known loss-of-function control p.Pro214Leu were established. Subcellular localization was assessed via immunofluorescence microscopy with quantitative scoring. Transcriptional repression was evaluated using luciferase reporter assays driven by ETV6 target promoters (MMP3 and PF4). p.Arg202Gly predominantly showed nuclear localization comparable to WT, whereas p.Pro214Leu was enriched in the cytoplasm. In the reporter assays, similar to WT, p.Arg202Gly retained repression activity. Meanwhile, p.Pro214Leu failed to repress both reporters. The in-silico prediction of nuclear export sequences suggested an additional export signal in p.Pro214Leu, but not in p.Arg202Gly. Collectively, these findings indicate that p.Arg202Gly behaves as WT-like in the assays performed and do not support a loss-of-function effect. Our study emphasizes the importance of variant-level functional assessment for rare ETV6 variants to inform clinical interpretation and avoid overestimation of pathogenicity.

Airway structural remodeling and hyperresponsiveness (AHR), hallmarks of asthma, are influenced by genetic variations and adverse exposures. While intrauterine perturbations in lung development have been linked to adult pulmonary disease, the developmental origins of these abnormalities remain poorly understood. Here, we provide evidence of genetic background playing a key role in this process. Using A/J and C57BL/6J mice known for their distinct susceptibility to AHR, we show that A/J embryos selectively develop an aberrant airway smooth muscle (SM) program and AHR in adulthood when exposed transiently to a vitamin A/retinoic acid (RA)-disrupted intrauterine environment in vivo by maternal BMS493 administration. Single-nucleus multiomics identified a mesenchymal cell population overactivating TGF-β targets in response to BMS493 selectively in A/J lungs. These cells, localized to sites of airway SM initiation and p-SMAD2- and -3, exhibited robust BMS493-mediated upregulation of SMAD2/3 targets, including regulators of SM program Pdgfra and Tnc. Functional analyses in vivo and cultured lungs showed aberrant SM formation in areas of overactive TGF-β of BMS493-exposed lungs. These abnormalities were prevented by inhibiting TGF-β signaling in utero in RA-deficient embryos. These findings underscore how distinct genetic backgrounds respond to intrauterine perturbations that program airway structure and function, with potential lasting consequences in postnatal pulmonary function.

In recent years ERphagy, the selective autophagic degradation of the endoplasmic reticulum (ER), has emerged as a key selective autophagic pathway involved not only in the recycling of the ER, but also in preventing the replication of viruses and bacteria. The mechanisms by which ERphagy achieves this do not seem immediately related to canonical xenophagy pathways and could provide a new avenue for therapeutic targets to combat pathogenic infections. In this editor's corner we briefly summarize the ways ERphagy is involved in pathogen infection, highlighting the potential ERphagy has as an understudied innate immune response pathway.<b>Abbreviation</b>: IFN-I: type I interferon; LPS: lipopolysaccharide; STING1: stimulator of interferon response cGAMP interactor 1.

<h4>Background</h4>Primary liver cancer, predominantly hepatocellular carcinoma (HCC), has limited therapeutic options. While mutation-derived neoantigen vaccine holds promise, its success is hindered by low antigen availability. This study explores transcriptome-derived neoantigens (neoantigen-encoding tumor-specific transcripts, neoTSTs) in HCC, characterizing their features, generation mechanisms, and therapeutic potential.<h4>Methods</h4>We developed a computational pipeline integrating STAR/StringTie-based transcript assembly with multiexon/single-exon reference datasets (23,972 human control samples) for tumor-specific transcripts (TSTs) identification. A custom sliding-window algorithm compared TST-encoded peptides against UniProt, with neoTSTs predicted using netMHCPan. This framework was applied to 1,013 patients with liver cancer. NeoTSTs were validated through proteomics, immunopeptidomics, and HLA-transgenic models. Multiomics analyses characterized splicing patterns, transposable elements, and transcription factor regulation. Single-cell RNA-seq and Hep53.4 murine models assessed tumor coverage and immunotherapeutic efficacy.<h4>Results</h4>We analyzed RNA-seq data from 1,013 patients with liver cancer and constructed a multilayered reference dataset. Using a customized pipeline, we identified an average of 60 neoTSTs per patient, significantly surpassing mutation-derived neoantigens (neoMuts). NeoTSTs exhibited higher population frequencies, with 73.1% providing multiple epitopes, and were validated through mass spectrometry and HLA transgenic mouse models. Mechanistically, neoTSTs were generated via retained introns, transposable element activation, HNF4A-regulated alternative promoters, and de novo transmembrane domain generation. Single-cell analysis revealed neoTSTs cover >75% of tumor cells and identified antigen-presenting cancer-associated fibroblasts that enriched in immunotherapy responders and amplified CD4⁺ T-cell responses. In murine HCC models, neoTST vaccination outperformed neoMuts, inducing dual major histocompatibility complex-I/II activation and significant tumor growth inhibition.<h4>Conclusions</h4>NeoTSTs represent a superior neoantigen source in HCC, compensating for the limitations of mutation-derived targets. The remarkable abundance and patient-to-patient sharedness of neoTSTs underscore their dual potential: (1) as personalized immunotherapeutic targets, and (2) as broadly applicable antigens for low-TMB tumors. These findings provide a transformative framework for expanding treatment options in HCC immunotherapy.

Neurodegenerative diseases (NDDs), including Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, and hereditary ataxias, remain major global health challenges with limited therapeutic options. Although clinically and genetically diverse, these diseases share extensively studied cellular and molecular hallmarks, including protein aggregation, impaired proteostasis, cytoskeletal abnormalities, altered energy metabolism, nucleic acid damage, and chronic inflammation. Emerging evidence indicates that dysfunction of the centrosome-cilium-satellite axis intersects with these established pathways in disease- and cell type-specific contexts. This axis, composed of centrosomes, primary cilia, and centriolar satellites, coordinates cytoskeletal organization, ciliary signaling, trafficking, proteostasis, and stress responses and acquires specialized functions in neurons that support polarity, connectivity, and long-term maintenance. In this review, we outline the structure, function, and neuronal specializations of the centrosome-cilium-satellite axis, then examine how its dysfunction has been reported in neurodegenerative disease models. We also discuss centriolar satellites as regulators of centrosome and cilium biology whose disease-specific roles in classical NDDs remain comparatively underexplored, with insights from Huntington's disease and schizophrenia. Finally, we discuss therapeutic strategies aimed at restoring axis structure and dynamics, modulating ciliary signaling, and correcting disease-linked genetic or transcript-level defects, emphasizing mechanism-based approaches that require validation in disease-relevant models. Together, the centrosome-cilium-satellite axis provides an emerging framework for understanding context-dependent organelle dysfunction in neuronal vulnerability and neurodegeneration.

<h4>Background</h4>Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal malignancy that is often diagnosed after curative treatment is no longer feasible. Existing biomarkers, particularly CA19-9, have limited sensitivity and specificity. Plasma proteins that capture tumor-associated biological alterations may therefore provide useful signals for earlier detection.<h4>Methods</h4>Plasma samples from 99 patients with PDAC and 30 healthy controls were analyzed using data-independent acquisition (DIA) proteomics. Differentially expressed proteins were identified using predefined statistical thresholds and further examined by functional enrichment analysis. Selected candidate biomarkers were validated by ELISA in an independent subset.<h4>Results</h4>Among 565 quantified plasma proteins, 52 were differentially expressed between PDAC and controls. These proteins were enriched in extracellular processes, cholesterol metabolism, complement and coagulation cascades, and pancreatic secretion pathways. ELISA validation confirmed higher plasma levels of Cathepsin S, CTRB2, MARCO, PIGR, PRDX6, REG1A, Trypsin-2, and PEP-FAP in patients with PDAC compared with healthy controls. ROC analyses showed moderate-to-good discriminatory performance for several candidates, and the MARCO + PEP-FAP model improved classification compared with either marker alone.<h4>Conclusion</h4>These findings reveal circulating proteins linked to key PDAC-related biological processes and identify eight candidates for further evaluation in multi-protein diagnostic panels. Larger validation studies incorporating clinically relevant disease control groups are warranted to determine their diagnostic specificity and clinical utility.

Aminoacyl-tRNA synthetases (ARSs) are a family of enzymes that attach amino acids to tRNAs. To date, all 37 human ARS genes have been implicated in genetic diseases, affecting over a thousand patients worldwide. At the 2025 Federation of European Biochemical Societies (FEBS) Special Meeting 'Expanding Frontiers in Aminoacyl-tRNA Synthetase Research', patients, families, and advocacy groups communicated the need for a unified approach for reporting on ARS gene names and associated conditions in the scientific literature. This request stemmed from the current use of multiple nomenclature systems and the desire of these individuals to rapidly identify published data on specific ARS genes. Here, we summarize ARS gene nomenclature and request that the scientific community adhere to a single nomenclature system.

<h4>Background</h4>Chronic pancreatitis (CP) progressively destroys pancreatic parenchyma and predisposes patients to pancreatogenic type 3c diabetes mellitus (T3cDM).<h4>Objectives</h4>We conducted a systematic review and meta-analysis of cohort studies enrolling adults with clinically and/or instrumentally confirmed CP and no pre-existing diabetes at baseline.<h4>Design</h4>Systematic review and meta-analysis of cohort studies.<h4>Data sources and methods</h4>Scopus, Web of Science Core Collection, and PubMed were searched from inception to August 2025. Random-effects models estimated pooled incidence of new-onset diabetes and the proportion requiring insulin. Prespecified subgroup analyses were stratified by follow-up duration (<60 months, 60-120 months, >120 months). Meta-regression explored demographic, behavioral, and clinical moderators.<h4>Results</h4>Nineteen studies met the inclusion criteria and contributed to the primary meta-analysis of incident diabetes after CP. The pooled incidence of diabetes was 30% (95% CI: 27-34; <i>I</i>² = 94.8%), with a prediction interval of 15%-49%. Exploratory follow-up-stratified analysis suggested a time-dependent increase: 18% (95% CI: 14-22) at <60 months, 25% (95% CI: 15-35) at 60-120 months, and 44% (95% CI: 24-66) at >120 months (<i>p</i> for subgroup differences = 0.0184). Among patients who developed diabetes, 59% (95% CI: 46-71; 7 studies; <i>I</i>² = 91.7%) required insulin therapy. Meta-regression did not identify statistically significant study-level moderators, although pancreatic calcification showed a non-significant positive trend.<h4>Conclusion</h4>CP is associated with a substantial and time-dependent incidence of diabetes; however, the pooled estimates should be interpreted cautiously because the included studies were observational and highly heterogeneous. These findings support long-term glycemic monitoring and careful classification of pancreatogenic diabetes in patients with CP, while optimal screening intervals and treatment timing require further prospective evidence.<h4>Trial registration</h4>PROSPERO CRD420251136931.

Neurodegenerative diseases represent a major and growing global health burden. Although these disorders are often clinically defined by symptoms and affected brain regions, many are mechanistically linked to abnormal protein accumulation, misfolding, impaired proteostasis, RNA dysregulation, mitochondrial dysfunction, and neuroinflammation. In this Perspective article, I discuss major neurodegenerative diseases, including Alzheimer's disease, Parkinson's disease, dementia with Lewy bodies, multiple system atrophy, amyotrophic lateral sclerosis, frontotemporal dementia, Huntington's disease, prion diseases, spinocerebellar ataxias, and spinal muscular atrophy, through the lens of disease-associated proteins and experimental modeling. I argue that a protein-centered framework provides a useful approach for understanding disease mechanisms and selecting transgenic mouse models, while recognizing that aging, cellular context, neuroinflammation, mitochondrial dysfunction, vascular dysfunction, and other disease modifiers also shape neurodegeneration. Transgenic and genetically engineered mouse models have been essential for dissecting the pathogenic roles of amyloid-β, tau, α-synuclein, TDP-43, SOD1, FUS, <i>C9ORF72</i>-associated dipeptide repeat proteins, mutant huntingtin, prion protein, ataxins, and SMN deficiency. However, these models have important limitations, including artificial overexpression, familial mutation bias, species differences, and incomplete representation of aging-related sporadic diseases. Rather than seeking a single "best" model, a more productive strategy is to adopt model portfolios tailored to specific biological questions and to integrate mouse studies with human cellular models, postmortem tissue, omics approaches, and biomarker-based validation. Such an approach may improve mechanistic insight, strengthen translational relevance, and enhance the predictive value of preclinical neurodegenerative disease research.

Thermoresponsive polymeric prodrugs represent a promising strategy for localized and sustained in vivo drug delivery. In this work, two polyethylene glycol (PEG)-based dexamethasone (Dex) prodrugs with different Dex contents were synthesized using a four-arm PEG scaffold. Prodrug <b>1</b>, containing four Dex molecules, showed high aqueous solubility but no thermoresponsive gelation behavior. In contrast, eight-Dex Prodrug <b>2</b> exhibited temperature-dependent aggregation and formed hydrogels in aqueous media. The viscosity of the Prodrug <b>2</b> hydrogel was reduced by introducing 10% ethanol as a cosolvent, enabling an injectable formulation that rapidly forms a hydrogel depot upon contact with aqueous media. The hydrogel provides gradual Dex release via the cleavage of the acid-labile hydrazone bond linking Dex to the PEG. In a monosodium iodoacetate (MIA)-induced osteoarthritis (OA) pain model, intra-articular (IA) injection of Prodrug <b>2</b> produced rapid and sustained pain relief for up to 28 days. These findings indicate that the hydrogel-forming four-arm PEG-based Dex prodrug offers a potentially effective approach for prolonged local corticosteroid (CS) delivery, applicable to the treatment of many local pathologies, including OA and OA pain.

Autoimmune myocarditis is an inflammatory disease of the myocardium driven by immune dysregulation and genetic predisposition. Recent advances in genomics, including genome-wide association studies (GWAS), have revealed key loci and pathways involved in disease susceptibility. This review synthesizes current knowledge on genetic determinants, from classical candidate genes to emerging GWAS findings, and explores their clinical implications for risk stratification and precision medicine.

Inherited liver diseases are predominantly caused by monogenic mutations, and the vast majority of these conditions currently lack curative treatment options. Although liver transplantation may be used for patients with end-stage disease, it faces numerous challenges, including donor organ shortage, immune rejection, and the need for lifelong immunosuppression. In recent years, CRISPR-Cas9-based gene editing technology has advanced rapidly, offering transformative hope for the treatment of these diseases. This review systematically elucidates the working principles and technical advantages of the CRISPR-Cas9 system and its derived tools (base editing and prime editing), summarizes recent applications of these technologies in the treatment of hereditary liver diseases, and discusses the prospects and challenges of their clinical translation, aiming to provide a theoretical reference for future research in this field.

Oral squamous cell carcinoma (OSCC) accounts for most head-and-neck cancers, and effective biological adjuvants remain limited. Gingival mesenchymal stem cells (GMSCs) exhibit anti-tumour paracrine activity, but the underlying molecular mechanisms and their relevance in patient cohorts remain incompletely understood. Consensus apoptosis-reactive oxygen species (ROS) effectors were identified through integrated transcriptomic analyses of TCGA-HNSC and three GEO cohorts. Candidate genes were evaluated in primary OSCC cells exposed to GMSC-conditioned medium or indirect Transwell co-culture. Findings were further examined using patient-cohort validation, single-cell ligand-receptor analysis, pathway and transcription-factor activity inference, and drug-repurposing approaches. Computational analyses identified an apoptosis-ROS network centred on <i>BAX</i>, <i>BCL2</i>, <i>CASP3</i>, <i>CASP9</i>, <i>NOX1</i>, and <i>GPX1</i>. Indirect GMSC co-culture reduced intracellular ROS, increased early apoptosis, and induced G2/M accumulation, whereas conditioned medium produced inconsistent effects, suggesting a requirement for live bidirectional paracrine signalling. <i>BAX</i> was the only consistently up-regulated effector. The axis demonstrated concordant differential expression across independent HPV-negative OSCC cohorts but was not independently prognostic under leakage-free cross-validation or external validation. Pathway analyses supported ROS suppression, apoptosis activation, and altered stromal-tumour communication. Drug-repurposing analyses identified HSP90 inhibitors and the FDA-approved TOP2 inhibitor mitoxantrone as candidate therapeutic agents. GMSC paracrine activity targets a biologically interpretable apoptosis-ROS axis in OSCC that is reproducibly expressed across patient cohorts but does not constitute an independent prognostic biomarker. The identified therapeutic candidates warrant further experimental investigation.

Respiratory medicine is transitioning from symptom-driven, standardized care to a more precise, patient-specific approach guided by molecular profiling. This evolution is being enabled by advances in liquid biopsy, multiomics, and artificial intelligence (AI) analytics. Fractional exhaled nitric oxide (FeNO) and blood eosinophils, the two commonly used markers in asthma, are now being joined by more precise airway markers such as galectin-10, which could aid clinicians in making more informed decisions for biological treatments. In chronic obstructive pulmonary disease (COPD) similar progress is underway, with treatment now emphasizing inflammation endotypes, especially eosinophilic patterns, to direct therapeutic choices. Alongside these developments, routine blood-based ratios (e.g., platelet-to-lymphocyte and neutrophil-to-lymphocyte) are being explored as predictors of exacerbation risk, and forced oscillation testing (FOT) is proving useful for picking up early disease shifts. In more severe conditions, biomarkers are linked to an early and better prognosis, enabling timely intervention. Markers like Matrix metalloproteinase-7 (MMP-7) and CC chemokine ligand 18 (CCL18) have proven to be reliable indicators of mortality and disease progression in idiopathic pulmonary fibrosis. Meanwhile, in lung cancer, liquid biopsies, especially those measuring circulating tumor DNA and micro-RNA (miRNA) panels, are enhancing screening accuracy while helping to cut down on the high false-positive rates seen with low-dose computerised tomography (CT). Other respiratory conditions such as bronchiectasis, pulmonary embolism, pneumonia, and acute respiratory distress syndrome (ARDS) are also benefiting from biomarker advances. At the same time there is a growing push to standardize how these biomarkers are measured. AI-based clinical decision support systems are also playing an increasingly important role in the translation of all these complicated data into actionable clinical insights. Together these developments pave the way for improved respiratory care that is precise and responsive to individual patient needs.

Keratins (KRTs) are intermediate filament proteins expressed in epithelial cells and serve as diagnostic cancer biomarkers. They serve pivotal roles in tumor progression and metastasis, but their prognostic value in lung adenocarcinoma (LUAD) and relationship with the tumor immune microenvironment remain unclear. Gene expression of KRTs in The Cancer Genome Atlas (TCGA)-LUAD was analyzed; candidate genes were selected using LASSO and random forest. An XGBoost classifier with SHAP interpretation distinguished tumor from normal tissues. Prognostic models were developed using 101 algorithms with 10-fold cross-validation and validated in two independent GEO cohorts. Immune cell composition and pathway activity were assessed by CIBERSORT, MCP-counter, ssGSEA and GSEA; the TIDE score estimated potential immunotherapy response. Functional validation of KRT81 was performed via shRNA knockdown in LUAD cell lines, followed by proliferation, apoptosis and migration assays. A KRT-based diagnostic and prognostic signature was established. The XGBoost model achieved high accuracy (TCGA AUC=0.996; GSE31210 AUC=0.860); SHAP identified KRT81 as a key contributor. A four-gene prognostic model (KRT27, KRT80, KRT16, KRT81) stratified patients into high- and low-risk groups. The risk score was associated with advanced tumor stage and independently predicted overall survival (multivariate HR=2.16, 95% CI 1.43-3.28, P=2.0×10-4). High-risk tumors enriched proliferation/stroma pathways (cell cycle, DNA repair, ECM-receptor interaction, focal adhesion, p53 signaling); low-risk tumors enriched immune pathways. Immune profiling revealed reduced T/B cell infiltration, increased endothelial cells and higher T-cell exclusion scores in high-risk patients, indicating an immunosuppressive microenvironment. Higher risk scores also associated with chemotherapy resistance. Functional validation confirmed that shRNA-mediated KRT81 knockdown reduced LUAD cell proliferation, migration and invasion, while promoting apoptosis. These findings link KRT expression to clinical outcomes, the tumor microenvironment and therapeutic response in LUAD, suggesting roles for KRTs in cancer progression, chemotherapy resistance and predictive potential for immunotherapy response. This study provides new insights for prognostic evaluation and therapeutic decision-making in LUAD.

Mitochondria are central regulators of cellular metabolism, redox balance, calcium signaling, and cell survival, making them essential for neuronal function. Because neurons rely heavily on mitochondrial oxidative phosphorylation to meet their high energetic demands, mitochondrial dysfunction has emerged as a key pathogenic driver in major neurodegenerative diseases, including Alzheimer's disease, Parkinson's disease, Huntington's disease, and amyotrophic lateral sclerosis. Defects in mitochondrial bioenergetics, excessive reactive oxygen species production, impaired mitochondrial dynamics, disrupted mitophagy, and dysregulated calcium handling collectively contribute to neuronal damage, synaptic dysfunction, and neuroinflammation. These insights have prompted growing interest in therapeutic strategies that directly target mitochondria to restore organelle homeostasis. Recent advances in chemical biology and nanomedicine have enabled the development of mitochondria-targeted ligands, peptide-based targeting systems, and carrier or nanotechnology-enabled delivery platforms designed to overcome biological barriers and selectively deliver therapeutic cargos to mitochondria within the central nervous system. In this Review, we summarize mitochondrial pathological mechanisms in neurodegenerative diseases and discuss emerging mitochondria-targeted therapeutic strategies, highlighting delivery technologies, therapeutic modalities, and translational challenges. Although most strategies remain at the preclinical or proof-of-principle stage, these advances are beginning to shape a conceptual framework for precision mitochondrial medicine, with the longer-term goal of developing disease-modifying interventions for neurodegenerative disorders.

<h4>Background</h4><i>CACNA1E</i> mutations cause developmental and epileptic encephalopathy. We report a Chinese pediatric patient with epilepsy and congenital heart disease who carries a novel CACNA1E variant.<h4>Methods</h4>Whole-exome sequencing (WES) was performed on the proband and his unaffected parents. The variant was validated by Sanger sequencing and screened in 200 healthy controls. Bioinformatic tools and ACMG criteria were used for pathogenicity assessment.<h4>Results</h4>A <i>de novo</i> heterozygous missense variant, NM_000721.4: c.1256G > A (p.R419Q), was identified. It was absent in both parents and in 200 controls. The variant lies in a highly conserved and intolerant region. ACMG classification: likely pathogenic (PS2 + PM2 + PP3). The patient showed marked short-term response to adrenocorticotropic hormone (ACTH).<h4>Conclusion</h4>This novel variant expands the mutational spectrum of CACNA1E and provides a potential treatment clue for ACTH responsiveness in CACNA1E-related spasms.

<title>Abstract</title> <p> <bold>Background</bold> Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a debilitating multisystem disorder characterised by profound fatigue, post-exertional malaise, muscle pain and weakness. Although skeletal muscle abnormalities and mitochondrial alterations have been reported in people with ME/CFS, the underlying biological processes remain poorly understood, and no validated disease models, biomarkers or effective treatments are currently available. This study used patient-derived induced pluripotent stem cell (iPSC) myogenic progenitor (MP) cells to investigate intrinsic transcriptomic abnormalities in ME/CFS and identify potential drug repurposing candidates. <bold>Methods</bold> MP cells were generated from iPSCs derived from people with ME/CFS and healthy controls, and successfully characterised. Global mRNA sequencing was performed to compare transcriptomic profiles between ME/CFS and control MP cells (n = 6 ME/CFS; n = 7 controls). Differentially expressed genes and pathways were identified, followed by drug repurposing analysis using the Library of Integrated Network-based Cellular Signatures drug database (LINCS2) and supporting literature review. <bold>Results</bold> RNA sequencing identified seven differentially expressed genes at adjusted p < 0.05, including <italic>MIR205HG</italic> , <italic>FEZF1</italic> , <italic>HLA-DMB</italic> , <italic>SLC1A2</italic> , <italic>SYT13</italic> , <italic>GALNT4</italic> , and <italic>SLC2A14</italic> . Pathway analysis identified 73 differentially expressed pathways, of which 96% were downregulated in ME/CFS MP cells. Downregulated pathways included those involved in cell cycle regulation, DNA replication, mismatch repair, immune response and muscle cytoskeleton regulation-related pathways. In contrast, upregulated pathways were associated with metabolic reprogramming, including increased reliance on branched-chain amino acids for energy production. These findings suggest intrinsic abnormalities in ME/CFS-derived MP cells, particularly involving metabolic regulation, mitochondrial function and muscle-related cellular processes. Based on the differentially expressed genes, LINCS2-based drug repurposing analysis and literature review identified 22 candidate drugs with potential relevance to ME/CFS. These included safe and widely available agents such as leflunomide, melatonin and midodrine, which target pathways related to antiviral defence, immunomodulation, neurotransmitter modulation, autonomic regulation and vascular function. <bold>Conclusions</bold> Patient-derived iPSC-MP cells provide a useful model for investigating intrinsic skeletal muscle-related abnormalities in ME/CFS. Transcriptomic profiling revealed altered pathways related to cell cycle regulation, immune function, cytoskeletal organisation and energy metabolism, supporting the involvement of mitochondrial and metabolic dysfunction in ME/CFS pathobiology. Drug repurposing analysis identified several clinically relevant candidates that may warrant further functional validation as potential therapeutic options for ME/CFS. </p>

Amoebiasis is a critical global parasitic disease caused by <i>Entamoeba histolytica</i>. It includes amoebic colitis and extraintestinal abscesses. <i>E. histolytica</i> peroxiredoxin (<i>Eh</i>Prx) is a crucial antioxidant enzyme that maintains redox homeostasis and participates in counteracting oxidative stress, supporting trophozoite survival and pathogenicity in the host. We observed the <i>Eh</i>Prx structure to be a decameric ring formed through the polymerization of five dimers. <i>Eh</i>Prx had a unique α-helical structure at its N-terminus. Protein-protein docking and surface plasmon resonance (SPR) assay results suggest that <i>Eh</i>Prx may bind to myeloid differentiation factor 2 (MD2), the ligand protein of toll-like receptor 4 (TLR4), rather than directly to TLR4. Furthermore, <i>Eh</i>Prx was crucial in the induction of cellular ferroptosis by <i>E. histolytica</i> trophozoites and regulated ferroptosis in host cells through the TLR4/MD2 pathway. These findings may hold a solid structural basis and promising concepts for investigating the pathogenic mechanisms and drug targets of <i>E. histolytica</i>.<h4>Importance</h4><i>Entamoeba histolytica</i> is the causative agent of amoebiasis, a significant global health concern. Within this parasite, <i>E. histolytica</i> peroxiredoxin (<i>Eh</i>Prx) plays a pivotal role in mitigating oxidative stress, a critical survival mechanism in the hostile environment of the human host. <i>Eh</i>Prx belongs to a family of antioxidant enzymes responsible for detoxifying peroxides, which are deleterious byproducts of cellular metabolism and host-derived immune responses. By neutralizing reactive oxygen species (ROS), EhPrx safeguards the parasite's cellular integrity, ensuring its survival, proliferation, and pathogenicity. Despite its functional importance, the precise structural details of <i>Eh</i>Prx remain elusive, hindering a comprehensive understanding of its molecular mechanisms. Cryo-electron microscopy (cryo-EM) offers a promising avenue for elucidating the high-resolution structure of <i>Eh</i>Prx, which could reveal critical insights into its biological functions and inter- or intramolecular interactions. Such structural characterization may be indispensable for advancing our knowledge of <i>E. histolytica</i> biology and identifying novel diagnostic markers to combat amoebiasis effectively.

Oral nanomedicines that modulate gut microbiota and gut-brain interactions are crucial for effectively treating inflammatory bowel disease (IBD) and associated psychiatric disorders, such as anxiety and depression. However, the underlying causes of psychiatric disorders in patients with IBD remain unclear, and effective treatment strategies have yet to be established. Herein, we developed oral gastroprotective βG@Apr-WPG NMs (β-glucan armored apremilast encapsulating tryptophan-poly(lactic-co-glycolic acid)-glutathione nanomicelles). This system effectively treats IBD and associated anxiety/depression by modulating the microbiota-gut-brain axis. βG@Apr-WPG NMs demonstrated gastroprotection enhances retention, and enables sustained release within the inflamed colon, enhances drug solubility, and inflammation-responsive apremilast release significantly improving oral therapeutic efficacy. Oral βG@Apr-WPG NMs administration outperformed free apremilast by restoring gut barrier integrity, reducing histopathological damage, and modulating microbial dysbiosis. Systemic inflammation and neuroinflammation were markedly suppressed. Notably, the βG@Apr-WPG NMs ameliorated anxiety- and depression-like symptoms, as well as cognitive deficits in colitis-induced mice, highlighting its therapeutic impact beyond the gut. Addressing reduced efficacy of conventional therapies, this multifunctional βG@Apr-WPG NMs platform offers safe, simple, and highly efficient therapeutic strategy integrating targets intestinal inflammation, microbiota-gut-brain axis modulation, in the pathogenesis of IBD with comorbid neuropsychiatric disorders with confirmed safety.

Renal arcuate vein thrombosis (RAVT) is a rare and often underrecognized cause of acute kidney injury (AKI) that typically requires renal biopsy for diagnosis. We report a 29-year-old man who developed AKI following heavy alcohol intake, an upper respiratory tract infection, and exposure to multiple medications including repeated nonsteroidal anti-inflammatory drugs and traditional Chinese patent medicines. Laboratory tests showed mild proteinuria without hematuria and negative autoimmune serology, with normal renal imaging. Renal biopsy revealed acute tubular injury and thrombus-like material within arcuate veins, accompanied by mild medullary hemorrhage and focal vascular necrosis, findings suggestive of arcuate venous thrombosis. Renal function recovered completely after supportive therapy within eight days.This case highlights that an arcuate venous lesion suggestive of thrombosis may be an underrecognized finding in patients with multifactorial, biopsy-proven AKI, even when the clinical presentation and imaging findings are nonspecific. Awareness of this possibility may broaden the differential diagnosis in selected patients with unexplained or disproportionate AKI and multiple renal risk factors, and kidney biopsy may help identify otherwise unrecognized vascular involvement.

The Karan Fries (KF), a newly developed dairy cattle (Holstein Friesian × Tharparkar), after nine generations of inter se mating (mating among crossbreds) by maintaining a stable, composite breed structure, is expected to preserve some heterosis or hybrid vigour through the retention of heterozygosity and selective pressure for performance traits. Although a decline in productivity is generally seen from F1 to later inter se generations, the rate of loss of heterosis slows down, allowing the population to reach a new equilibrium. This seems to be an interesting proposition. Therefore, to assess the retained heterosis and to evaluate the current status of the KF population, the present study was conducted, which included a sizeable phenotyped and genotyped animals (n = 355). The analysis identified a stabilized genomic architecture with a mean exotic inheritance of 64.34 ± 0.74% (predominantly Holstein Friesian) and an indigenous contribution of 35.66 ± 0.95% (primarily Tharparkar). Genomic indicators of heterosis revealed a mean Genomic Retained Heterosis (RH_g) of 59% which accounts for 15.82% of the total observed KF average yield (4153.14 ± 167.32 kg). Genomic Retained Heterozygosity (RHET_g) of 0.44%. Notably, a near-perfect correlation (r = 0.98) was observed between these two metrics, indicating that RHET_g can be used interchangeably with RH_g to assess heterosis and its effects in instances where parental genotypic records were absent. Linear regression analysis demonstrated that RH_g was a primary driver of productivity, with Total Milk Yield (TMY) increasing by 1113.81 ± 301.91 kg per unit increase in RH_g (p < 0.001). The impact on TMY was significantly larger than on 305-day milk yield (339.08 ± 282.92 kg), suggesting that genomic heterosis specifically enhances lactation persistence and environmental robustness. A joint-model GWAS identified 182 SNPs with significant additive effects and 117 SNPs with significant dominance effects for TMY. Functional annotation revels candidate genes for milk synthesis (SLC25A1, PIP4K2A, LATS2), heat stress resilience (DNAJB5, DNAJC1), and immunological defence (ARHGAP15, LPAR3). Protein network analysis identified the Mitochondrial Ribosomal Protein (MRP) family, specifically hub gene MRPL22, as the central driving gene for both additive and dominance components. These findings confirm that KF cattle effectively leverage retained heterosis to integrate high production potential with robust tropical climatic adaptability.

<h4>Background</h4>Alzheimer's disease (AD) is a degenerative central nervous system disorder characterized by progressive cognitive and behavioral impairment. As nanoscale intercellular communication vesicles that carry AD-related pathological molecules, exosomes are promising biomarkers and therapeutic carriers for AD. In this study, we downloaded AD-related gene expression profiles and clinical data from the Gene Expression Omnibus (GEO) database (datasets GSE138260, GSE29378, GSE36980, and GSE5281). Through a series of bioinformatics analyses, clinical predictive model construction, pharmacological network analysis, and molecular docking simulations, we developed an exosomal gene-based predictive model for AD pathogenesis and identified potential pharmacological networks and molecular docking targets for AD treatment.<h4>Materials and methods</h4>AD-related gene expression and clinical data were retrieved from the GEO database. Bioinformatics analyses, clinical model construction, drug-gene network analysis, and molecular docking were subsequently performed to explore exosomal gene models for predicting AD pathogenesis, as well as potential pharmacological networks and molecular docking targets for AD therapy.<h4>Results</h4>A five-exosomal-gene predictive model was established, comprising CD44, CXCR4, TUBB, PSMA5, and PSMB3. Pharmacological network analysis of these five genes revealed their significant associations with chelidonine, 2-chloro-1,4-dinitrobenzene, oxazolone, phencyclidine, thioridazine, and etodolac. Further molecular docking simulations identified key binding targets, including R41, Y42, R78, Y79, C77, I88, C97, A98, I96, I72, L70, E67, G103, I91, and T102.<h4>Conclusions</h4>Our comprehensive analyses successfully established a reliable exosomal gene-based model for predicting AD pathogenesis, and identified relevant pharmacological networks and core molecular docking targets, providing novel insights for AD diagnosis and targeted therapy.

Neurodegenerative diseases are progressive disorders that involve the loss and dysfunction of neurons. Alzheimer's disease, Parkinson's disease, Amyotrophic lateral sclerosis, Huntington's disease, Frontotemporal dementia are examples of diseases. While different clinically, these disorders have a common genetic, molecular and cellular basis. This review examines the common genetic pathways, along with the interactions between genes of major neurodegenerative diseases, with a focus on the key genes, such as APOE, SNCA, MAPT, TARDBP, LRRK2 and HTT. The common pathogenic mechanisms considered to play a major role in disease progression include protein misfolding and aggregation, mitochondrial dysfunction, oxidative stress, neuroinflammation, diminished autophagy, and impaired lysosomal function, as well as synaptic degeneration. The review also emphasizes the role of systems biology strategies, such as genome-wide association studies, transcriptomics, proteomics, metabolomics, interactome analysis, and multi-omics integration, to unveiling complex molecular networks in neurodegeneration. Furthermore, the emerging biomarker strategies and therapeutic strategies targeting convergence signaling pathways including NF-κB, PI3K-Akt-mTOR, MAPK and Wnt/β-catenin are summarized. The common genetic basis and the cross-connecting molecular mechanisms of the various neurodegenerative diseases could help in the discovery of new biomarkers and pan-therapeutic targets. Further advances in molecular genetics, computational biology and precision medicine are needed to enhance early detection and the creation of effective disease-modifying treatments.

The Angiotensin II type-2 receptor (AGTR2) is a G protein-coupled receptor (GPCR) that mediates vasodilatory, anti-proliferative, and cardioprotective responses as part of the renin-angiotensin system (RAS). However, the precise structural dynamics underlying its ligand-specific activation remain incompletely understood. In this study, we performed long-timescale molecular dynamics (MD) simulations to investigate how Angiotensin peptides differentially modulate the structural dynamics of AGTR2 in both inactive- and active-like conformations and to elucidate ligand-specific modulation of AGTR2 in the RAS protective arm. Ang II stabilized a 'Locked Active State' through compact TM3-TM6 distances, persistent hydrogen bonding (PHE8-LYS215^5.42), salt bridges (ARG2-ASP279^6.58, ARG2-ASP297^7.32), hydrophobic contacts (TRP100^2.60, MET128^3.36), and rotameric locking of micro-switches. These interactions stabilized Helix 8, enhancing dynamic network connectivity and supporting ligand-dependent activation-related conformational tendencies rather than a full canonical GPCR activation transition. Conversely, Ang1-7 induced a 'Flexible Intermediate State' characterized by weaker interactions (PRO7-THR125^3.33), dynamic instability at the Helix 8 interface, and increased conformational sampling. Activation motif analyses (CWxP, PIF, E/DRY, NPxxY), DCC maps, CP, DRIN metrics, and PCA confirmed distinct signaling features for each ligand. This study provides a dual functional profile for AGTR2, where Ang II acts as a strong conformational stabilizer promoting activation-related dynamic features, while Ang 1-7 serves as a dynamic modulator. These findings contribute to a structural and dynamic framework for understanding AGTR2 signaling and supporting its therapeutic potential in fine-tuning cardiovascular responses within the RAS.

Patient-derived organoids (PDOs) can recapitulate selected features of original tumors and provide a useful system for studying epithelial ovarian cancer (EOC) in three-dimensional culture. Nonetheless, the protocol to generate EOC-PDOs has not yet been standardized. We therefore empirically refined the culture conditions and medium composition, considering the protocol effective when EOC PDOs were successfully established in five consecutive attempts. Tissue and ascites samples (from 29 and 8 patients, respectively) were used and the Syringe-extruded Organoid Basement membrane extract Assembly (SOBA) fragment culture method applied. Selected similarities in cell morphology, marker expression, and detectable alterations between organoids and patient-matched tumor tissue were assessed using pairwise comparisons. The refined workflow used a culture medium containing recombinant human R-Spondin 1 protein (250 ng/mL), Noggin, and NRG1, factors implicated in organoid maintenance and expansion. Ultimately, 31 EOC organoid lines successfully generated from 82.8% (24/29) of tissue and 87.5% (7/8) of ascites samples. The SOBA technique increased the 10 day PDO yield by 2.75-fold relative to the surface-attached dome method (p < 0.0001). This empirically refined workflow can be adjusted for specific experimental applications; however, further validation across larger and clinically diverse EOC cohorts is required.

Gallbladder cancer (GBC), a lethal malignancy of the biliary tract, is associated with a poor clinical prognosis. Although chemo-immunotherapy combinations demonstrate preliminary efficacy, the molecular determinants of treatment response remain elusive. Emerging evidence implicates aberrant alternative splicing in modulating tumor immunity. Through an in vitro CRISPR/Cas9 screen, we identified SRSF2 as a key RNA-binding protein regulating PD-L1 expression. Intriguingly, SRSF2 does not directly bind PD-L1 mRNA. Multi-omics analyses (mRNA-seq, RIP-seq, and proteomics) revealed that SRSF2 induces exon skipping in hnRNPD, shifting isoform expression from full-length P45 to truncated P40. Functional studies established that P45-but not P40-binds to AU-rich elements in the PD-L1 3'-UTR to promote mRNA degradation. Leveraging this mechanism, we designed splice-switching antisense oligonucleotides (ASOs) that block SRSF2-mediated exon skipping, restoring P45 expression. This intervention effectively reduced PD-L1 levels and potentiated T-cell-mediated cytotoxicity in vitro and in vivo. These findings elucidate a splicing-centric mechanism of immune evasion and highlight the therapeutic potential of splicing modulation in cancer immunotherapy. Proposed model of the SRSF2-hnRNPD-PD-L1 axis in gallbladder cancer (GBC) immune evasion and its therapeutic targeting. Overexpression of SRSF2 drives hnRNPD exon skipping, shifting the isoform balance from the PD-L1-degrading P45 to the truncated P40. This transition stabilizes PD-L1 mRNA and facilitates tumor immune evasion. Conversely, therapeutic intervention with splice-switching ASOs blocks SRSF2-mediated alternative splicing, restores P45 expression, and effectively reactivates T-cell-mediated cytotoxicity against GBC cells.

<h4>Background</h4>Chagas disease (CD) is caused by Trypanosoma cruzi. Treatment is based on benznidazole (Bz), although it has significant limitations, such as low efficacy in the chronic phase. Therefore, the search for new therapies with greater selectivity and antiparasitic activity is necessary. In this context, 1,2,4-oxadiazole stands out for its biological properties, including antiparasitic activities.<h4>Objective</h4>To evaluate the in vitro activity of N-cyclohexyl 3-(3-methylphenyl)-1,2,4-oxadiazol-5-amine derivatives against the Y strain of T. cruzi and an in vivo toxicity study.<h4>Methods</h4>Cytotoxicity was evaluated in LLC-MK2 cells by the MTT assay, while the antiparasitic effect on the three T. cruzi life forms was determined by counting. Flow cytometry analyses were then conducted to investigate possible death pathway mechanisms and antioxidant and antiacetylcholinesterase activities. Scanning electron microscopy (SEM) was also performed to observe morphological changes caused by the compounds. Finally, in vivo acute toxicity tests were performed on ZebraFish embryos.<h4>Results</h4>The results presented show distinct cellular toxicity profiles in LLC-MK2 cells, in addition to demonstrating antiparasitic activity at different concentrations. In amastigotes, cytotoxic effects were stimulated. The molecules also induced an increase in reactive oxygen species and membrane damage, in addition to loss of integrity and morphological changes. The antioxidant activity revealed a high capacity for scavenging free radicals, suggesting an alteration of the redox balance of the parasite, in addition to showing inhibition of acetylcholinesterase, an important enzyme present in the formation of parasites, which choline is a constituent. In the ZebraFish model, molecule 2a showed dose-dependent embryonic toxicity, with an LC50 of 14-15 µM.<h4>Main conclusions</h4>The calculated conclusions appear to indicate an antiparasitic effect associated with cell death mechanisms. However, further studies are needed to reduce toxicity in the animal model and increase delivery to the site of action.

Huntington's disease is an inherited neurodegenerative disorder caused by a CAG repeat expansion in exon 1 of the huntingtin (<i>HTT</i>) gene, encoding an expanded polyglutamine tract in the huntingtin (HTT) protein. The pathogenic CAG repeat of <i>HTT</i> is unstable and undergoes progressive somatic expansion in specific brain cells and peripheral tissues throughout life. Genes involved in DNA mismatch repair pathways, which promote repeat expansion, have been identified as genetic modifiers of the disease. Consequently, the rate of CAG repeat expansion is a key determinant driving the age of onset and disease progression. As the CAG repeat expands, alternative processing of <i>HTT</i> pre-mRNA increasingly favours production of the <i>HTT1a</i> transcript, which encodes the highly pathogenic and aggregation-prone HTT1a protein. This process provides a mechanistic link between CAG repeat expansion and disease pathogenesis, as increased HTT1a production accelerates HTT aggregation and neuronal dysfunction. HTT1a has previously been detected in Huntington's disease mouse models by using immunoprecipitation coupled with western blotting, homogeneous time-resolved fluorescence (HTRF) and Meso Scale Discovery (MSD) bioassays and immunohistochemistry. These approaches were developed using MW8, a neoepitope antibody that specifically recognizes the C-terminus of HTT1a. MW8 is a relatively weak antibody with limited detection sensitivity. To generate more robust HTT1a-specific reagents, two novel recombinant antibodies, 1B12 and 11G2, have been developed for evaluation. Using an allelic series of knock-in (<i>Hdh</i>Q20, <i>Hdh</i>Q50, <i>Hdh</i>Q80, <i>Hdh</i>Q111, CAG140 and zQ175) mice, alongside transgenic YAC128 and N171-82Q models, we extensively evaluated and compared the performance of MW8, 1B12 and 11G2. We demonstrate that 1B12 and 11G2 function as HTT1a-specific neoepitope antibodies by immunoprecipitation with western blotting and by immunohistochemistry. To enhance HTT1a detection using HTRF and MSD technology platforms, we further evaluated the performance of 1B12 and 11G2 in HTT bioassays using cortical lysates from zQ175 and YAC128 mice. In zQ175 mice, enhanced detection of aggregated HTT1a by HTRF and MSD revealed that HTT fragments longer than HTT1a can be incorporated into HTT1a-containing aggregates. The most sensitive assays were subsequently applied across the allelic series of knock-in mice to assess the effect of polyglutamine length on bioassay performance. For optimal sensitivity, we recommend the preferential use of 1B12 for HTRF assays and 11G2 for MSD assays. Collectively, these findings establish 1B12 and 11G2 as robust antibodies to reliably detect and track HTT1a pathology <i>in vivo</i> and promote the replacement of previously used MW8-based experimental approaches.

<h4>Background</h4>Gastric cancer (GC) progression entails profound epithelial-stromal remodeling, yet how epithelial metabolic programs are linked to myeloid-dominated immune exclusion and prognostic risk remains unclear. This study aimed to integrate single-cell RNA sequencing (scRNA-seq) and bulk transcriptomic analyses to characterize a lactate-myeloid-related risk axis in GC.<h4>Methods</h4>We integrated six scRNA-seq datasets comprising 116 samples and 355,091 cells spanning paratumor (PT), chronic atrophic gastritis (CAG), moderate intestinal metaplasia (MIM), severe intestinal metaplasia (SIM), early gastric cancer (EGC), and advanced gastric cancer (AGC), and coupled them to bulk cohorts. Epithelial and myeloid states, pseudotime trajectories, ligand-receptor programs, and pathway activities were analyzed. A 19-gene prognostic model anchored on SLC16A3/MCT4 co-expression and macrophage immunoregulatory modules was developed in The Cancer Genome Atlas stomach adenocarcinoma (TCGA-STAD) cohort and externally validated in GSE84437. Mutation burden, tumor immune dysfunction and exclusion (TIDE), and expression-based drug-response predictions were further evaluated. SLC16A3 expression was validated by immunohistochemistry (IHC) in an independent clinical cohort.<h4>Results</h4>The integrated atlas showed immune contraction with epithelial expansion in premalignant lesions, followed by enrichment of aberrant epithelium and myeloid lineages in EGC/AGC. Within epithelium, a late-pseudotime SLC16A3+ lactate-export program increased with progression and was associated with extracellular matrix (ECM) remodeling, pro-angiogenic signaling, and myeloid-recruiting chemokines. IHC confirmed marked SLC16A3 enrichment within dense, cohesive neoplastic nests of Lauren intestinal and mixed subtypes. Myeloid decomposition revealed macrophage-centric expansion dominated by SPP1+ and APOE+ tumor-associated macrophage (TAM) states. The model-derived risk score stratified overall survival independently of clinical covariates. Low-risk tumors exhibited higher mutation counts and tumor mutational burden (TMB), whereas high-risk tumors were genomically quieter but TIDE-high. Expression-based analyses nominated differential drug-response patterns across risk groups, which require further validation.<h4>Conclusions</h4>This study identifies an epithelial SLC16A3-associated lactate-myeloid transcriptional program linked to macrophage-rich immune exclusion and poor prognosis in GC. The 19-gene model provides a biologically interpretable framework for risk stratification, while the immune-evasion and drug-response findings should be considered hypothesis-generating and require prospective and functional validation.

Background Cognitive impairment (mild cognitive impairment and early dementia) is a growing concern with limited therapeutic options. L-arginine, a semi-essential amino acid and precursor of nitric oxide, has shown neuroprotective effects in animal studies, but human trials, particularly in India, are lacking. This pilot study aimed to assess the safety and efficacy of oral L-arginine in elderly patients with cognitive impairment. Methodology This was a prospective, double-blind, randomized, placebo-controlled pilot study conducted at a tertiary care hospital in North India. Sixty-six subjects aged ≥60 years with subjective cognitive complaints were screened using the Hindi version of Addenbrooke's Cognitive Examination III (ACE-III); those scoring ≤82 were included after excluding reversible causes, including low vitamin B12 levels and abnormal thyroid-stimulating hormone (TSH) levels, coronary artery disease (CAD), and substance use. Thirty-eight participants were randomized to receive either oral L-arginine 6.4 g/day or a matching placebo for 30 days. Cognitive function was reassessed post-intervention using ACE-III. Adverse effects and tolerability were recorded. Results Thirty-five subjects completed the study (16 L-arginine and 19 placebo). Baseline characteristics were comparable between groups. After 30 days, the L-arginine group showed statistically significant within-group improvements in attention (p=0.018), memory (p<0.001), and language (p=0.017) domains, while the placebo group improved only in memory (p=0.003). Between-group comparison showed that significantly more patients in the L-arginine group achieved normal ACE-III scores (>82) compared to placebo (37.5% vs. 5.3%, p=0.032). Adverse effects leading to withdrawal in the L-arginine group included thyroid dysfunction (one participant), nausea/diarrhea (one participant), and bitter taste (one participant); no withdrawals occurred in the placebo group (p=0.353). Conclusions Oral L-arginine (6.4 g/day for 30 days) appears safe and improves attention, memory, and language domains in elderly patients with cognitive impairment. Larger, longer-term trials are warranted to confirm these findings and assess progression to dementia.

To address the pharmacokinetic limitations and moderate potency of current prostanoid IP receptor agonists for treating pulmonary arterial hypertension, we developed a series of conformationally constrained diphenylpyrazinyl amino cycloalkoxy acetic acids. By replacing the flexible side chain of MRE-269 with a rigid cyclic framework, we locked the bioactive conformation, identifying <b>7a-5</b> and its deuterated analog <b>7a-19</b>. Compound <b>7a-19</b> exhibited potent antiaggregatory activity (IC₅₀ = 0.97 μM), a 7-fold enhancement over MRE-269 (IC₅₀ = 7.4 μM). Crucially, leveraging the deuterium kinetic isotope effect in rats, <b>7a-19</b> demonstrated an 11-fold increase in systemic exposure (AUC _<i>o‑t</i> = 26.7 h μg/mL), a prolonged half-life (<i>t</i> _1/2 = 12.0 h), and excellent oral bioavailability (<i>F</i> = 47.3%). Combined with a clean safety profile in the Mini-Ames assay and a lack of hERG inhibition (IC₅₀ = 143.1 μM), <b>7a-19</b> stands out as a robust, long-acting lead candidate for PAH therapy.

<title>Abstract</title> <p> Adult stem cells maintain tissue homeostasis and regenerative capacity throughout life, but the mechanisms that preserve their integrity during aging and stress remain incompletely understood. Hair follicle pigmentation provides a tractable model to study stem cell maintenance as it relies on melanocyte stem cells (McSCs) whose depletion leads to age-related hair greying. Here, we identify the transcription factor BRN2 ( <italic>POU3F2</italic> ) as a key regulator of McSC homeostasis. We show that BRN2 activity is enriched in McSCs and declines with age in both mouse and human hair follicles. Using a melanocyte-specific conditional knockout mouse model, we reveal that loss of Brn2 promotes melanocyte differentiation and accumulation in the hair follicle bulb, accompanied by increased expression of melanocytic genes and a marked downregulation of DNA repair pathways, particularly those involved in nucleotide excision repair and chromosome maintenance. Brn2-deficient cells displayed impaired activation of the DNA damage response following ionizing radiation and entered a p53–p21–associated senescence-like differentiation state rather than undergoing apoptosis. In vivo, Brn2-deficient mice exposed to irradiation-induced stress developed rapid and permanent hair depigmentation associated with depletion of melanocyte lineage cells. Together, these findings identify BRN2 as a suppresor of stress-induced hair greying that links genome maintenance to melanocyte stem cell fate. </p>

Approximately half of Japanese patients with lung adenocarcinoma (LUAD) have epi-dermal growth factor receptor (<i>EGFR</i>) gene mutations. Therefore, <i>EGFR</i> is the most critical therapeutic target in treating LUAD. This study's purpose is to explore novel therapeutic targets for LUAD and further improve EGFR inhibitor combination therapy. We created microRNA (miRNA) expression signatures from clinical specimens of LUAD patients harboring <i>EGFR</i> gene mutations using RNA sequencing. Based on the miRNA signature, we focused on <i>miR-206</i>, which had the most downregulated expression. We searched for its target gene in LUAD cells by facilitating transfection of <i>miR-206</i> into the <i>EGFR</i> mutant cell line PC9 and searching for genes with suppressed expression. A total of 821 genes were downregulated in PC9 cells. Through molecular classification of these genes, we revealed that 18 genes were closely related to the cell cycle. Among these genes, we focused on cyclin-dependent kinase 4 (<i>CDK4</i>) and investigated the synergistic effects of a CDK4/6 inhibitor (abemaciclib or palbociclib) and osimertinib, an EGFR inhibitor in LUAD cells. In vitro and three-dimensional culture analyses showed that combination therapy with a CDK4/6 inhibitor and an EGFR inhibitor synergistically suppressed proliferation of LUAD cells with <i>EGFR</i> mutations. Our miRNA-based analysis is an excellent strategy for identifying therapeutic targets for LUAD. Continued analysis will reveal target molecules that enhance the therapeutic effects of EGFR inhibitors.

Uncoupling protein 1 (UCP1) is a mitochondrial inner-membrane carrier classically recognized as the molecular effector of non-shivering thermogenesis in brown adipose tissue. By dissipating the proton-motive force generated by oxidative phosphorylation, UCP1 converts stored chemical energy into heat and enables adaptive thermogenesis during cold exposure. The rediscovery of metabolically active brown adipose tissue (BAT) and inducible beige adipocytes in adult humans has renewed interest in UCP1-positive thermogenic adipose tissue as a regulator of systemic metabolism and a potential target for therapeutic modulation in obesity and cardiometabolic disease. Beyond thermogenesis, accumulating evidence indicates that UCP1-positive brown/beige adipocytes and thermogenic adipose tissue are involved in lipid and glucose metabolism, mitochondrial redox homeostasis, inflammatory remodeling, organ protection, and tumor-associated metabolic adaptation, mainly through adipocyte-autonomous mechanisms and adipose-organ communication. However, UCP1 biology is complex: BAT activity measured by imaging does not directly quantify UCP1 proton conductance, non-adipose UCP1 expression is often low and technically challenging to validate, local cell-autonomous UCP1 function in non-adipose tissues remains controversial, and UCP1-independent thermogenic pathways may compensate in selected contexts. In this review, we summarize the molecular and structural basis of UCP1 function, its regulation at transcriptional and post-transcriptional levels, and its biological roles in cellular and systemic homeostasis, with explicit distinction between direct adipocyte-autonomous UCP1 functions, indirect systemic effects mediated by thermogenic adipose tissue, and preliminary or incompletely validated evidence of local UCP1 activity in non-adipose cells. We further discuss the association of UCP1-positive thermogenic adipose tissue with obesity, type 2 diabetes mellitus (T2DM), cardiovascular disease, kidney injury, liver disease, neurological disorders, and cancer. Finally, we evaluate UCP1-related thermogenic adipose tissue activity as a biomarker and therapeutic target, highlighting current limitations, safety concerns, and future directions for precision metabolic medicine.

<h4>ABSTRACT</h4> Volume-regulated anion channels (VRACs), formed by leucine-rich repeat-containing 8 (LRRC8) proteins, are ubiquitously expressed chloride channels essential for cell volume regulation and implicated in diverse physiological and pathological processes. Small-molecule VRAC inhibitors have been reported to modulate paracrine signaling, proliferation, differentiation, migration, and apoptosis, and have been patented for potential therapeutic applications in stroke, cardiovascular and metabolic diseases, and cancer. However, growing evidence indicates that many commonly used VRAC blockers exert substantial off-target effects and frequently fail to reproduce phenotypes observed after deletion of the essential VRAC subunit LRRC8A. Here, we systematically compared effects of several widely used pharmacological VRAC inhibitors with outcomes of molecular downregulation of LRRC8A in limiting proliferation of malignant glioblastoma cells derived from surgical specimens. NIH/3T3 fibroblasts served as a non-malignant control. In serum-containing media, structurally diverse VRAC blockers (DCPIB, DIDS, carbenoxolone, phloretin, and bromadiolone) reduced proliferation in a non-uniform manner, with potencies that did not correlate with reported VRAC affinities and varied markedly among cell lines. Radiotracer-based measurements of VRAC activity indicated that these discrepancies were largely attributable to binding of inhibitors to serum albumin. When experiments were repeated under serum-free conditions, all inhibitors except DIDS and phloretin induced extensive death of both malignant and non-malignant cells, confirmed by microscopy and LDH release assays. This cytotoxicity was accompanied by a marked reduction in intracellular ATP levels, consistent with previously reported mitochondrial uncoupling effects. In contrast, LRRC8A knockdown reduced proliferation without substantial cell death. Together, these findings demonstrate that most commercially available VRAC blockers limit proliferation and viability predominantly through VRAC-independent mechanisms. Under standard culture conditions, serum albumin masks much of their intrinsic cytotoxicity. These results underscore the need for rigorous molecular controls in pharmacological studies and provide basis for developing more selective and less toxic VRAC-targeting agents.

Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by the emergence of autoantibodies and deposition of immune complexes. SLE presents with heterogeneous multi-organ involvement that varies among patients and its mechanisms have been investigated to facilitate appropriate stratification and treatment selection. Bulk RNA-seq and bulk ATAC-seq have provided important insights into the pathogenesis of SLE; however, these approaches are inherently limited by their reliance on predefined cell subsets and known markers, which can introduce bias and restrict their ability to fully resolve cellular heterogeneity. Recent advances in multi-omics analyses have enabled the investigation of multi-layered information beyond single-cell RNA sequencing (scRNA-seq) alone and have contributed substantially to elucidating the pathogenesis of SLE. Although the emergence of autoreactive B cells and the production of autoantibodies in SLE are well established, the mechanisms of evasion from negative selection remain unclear. Multi-omics analyses have revealed key aspects of SLE pathogenesis, particularly the expansion of atypical B cells (ABCs), an autoreactive population driven by extrafollicular pathways. Furthermore, beyond transcriptional profiling, multi-omics analysis has emerged as an additional investigative method, which combines scRNA-seq with other modalities. Spatial analyses, for instance, have provided critical insights into the tissue localization and persistence of autoreactive B cells within inflammatory niches in lupus nephritis, suggesting that local microenvironments contribute to treatment resistance. Additionally, B cell receptor (BCR) sequencing has revealed distinctive BCR features in SLE, such as reduced somatic hypermutation, increased repertoire naiveness, and increased immunoglobulin variable region heavy chain gene (IGHV4-34) usage in B cells in bone marrow and affected organs (not only in peripheral blood). Furthermore, emerging multimodal approaches integrating spatial, transcriptomic, and epigenomic information further highlight pathogenic cell-cell interactions and inflammatory circuits that cannot be captured by scRNA-seq alone. In this review, we summarize recent multi-omics studies that elucidate the origin, differentiation, and tissue localization of pathogenic B cells in SLE. We provide an overview of the multi-omics analyses focusing on B cells so far, especially at single-cell resolution, and discuss their possible applications in precision medicine.

<h4>Purpose</h4>This systematic review aims to provide an updated overview of the relative contribution of germline genetic and environmental factors to the pathophysiology of sagittal craniosynostosis (sCS).<h4>Methods</h4>Using the PubMed database in accordance with Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, relevant studies addressing genetic and/or environmental determinants of sCS in paediatric patients were systematically reviewed.<h4>Results</h4>A total of 1238 records were identified, of which 97 studies met inclusion criteria after full-text review. Overall, 25,877 patients were included, with 11,086 diagnosed with sCS (42.8%). Germline genetic variants potentially associated with craniosynostosis were reported in 251 patients, accounting for 12.6% of genetically tested individuals with sCS. Identified variants were distributed across 125 genes, with recurrent findings in 25 genes. Environmental and perinatal factors were more consistently reported across large population-based and case-control studies. Frequently associated factors included male sex, advanced maternal age, maternal smoking, thyroid dysfunction, fertility treatments, foetal constraint, prematurity, and abnormal birth weight, although effect sizes varied across studies.<h4>Conclusion</h4>Identifiable germline genetic causes appear to account for only a minority of sCS cases, whereas epidemiological evidence suggests that environmental and perinatal influences may also contribute to disease pathogenesis. Nevertheless, most cases lack identifiable germline mutations in key signalling pathways, and the available evidence does not support either genetic or environmental factors as individually deterministic drivers of disease development. Overall, the evidence synthesized in this review supports a multifactorial model for sCS, in which rare genetic susceptibility and non-genetic influences likely interact in the etiopathogenesis of the condition rather than reflecting a single dominant aetiology. However, heterogeneity across studies, differences in genetic testing methodologies, potential bias in exposure assessment, and the limited availability of integrative analyses restrict definitive conclusions regarding the relative contribution of these factors.

Faithful DNA replication requires precise control of replication fork progression to maintain genome integrity; yet, the mechanisms that restrain excessive fork acceleration remain unclear. We identify SUDS3 as a condensate-associated regulator of replication speed. SUDS3 forms dynamic nuclear condensates during S phase and under replication stress. These condensates spatially partition replication-associated factors, particularly MCM10, thereby limiting their accessibility to replication-associated chromatin. Loss of SUDS3 disrupts this spatial regulation and leads to aberrantly accelerated fork progression. Under replication stress, unchecked fork acceleration in SUDS3-deficient cells results in defective fork protection, excessive single-stranded DNA accumulation, ATR-CHK1 hyperactivation, and increased genome instability. Consequently, SUDS3 deficiency sensitizes cells to replication-targeting chemotherapeutic agents, a phenotype rescued by wild-type SUDS3 but not by condensate-defective mutant. Together, our findings reveal a condensate-based mechanism that constrains replication dynamics and establish SUDS3 condensates as critical safeguards of genome stability and potential vulnerabilities in replication-stressed cancers.

Histone deacetylases (HDACs) correlate closely with tumor prognosis and serve as therapeutic targets for malignancies. This study aimed to explore esophageal carcinoma (ESCA) therapeutic strategies via identifying histone deacetylation-related prognostic genes. We analyzed The Cancer Genome Atlas (TCGA)-ESCA and GSE53622 datasets, along with 17 histone deacetylation-related genes (HDRGs). Differentially expressed genes (DEGs1) between tumor and normal TCGA-ESCA samples were identified; DEGs2 linked to HDRGs were screened via unsupervised clustering of TCGA-ESCA tumors. Candidate genes were defined as the intersection of DEGs1 and DEGs2, followed by functional enrichment analysis and regression analyses. TCGA-ESCA samples were stratified into high-/low-risk groups, with model validation. Independent prognostic analysis, gene set enrichment analysis (GSEA), immune/gene mutation/drug sensitivity analyses, regulatory network construction, and RT-qPCR validation were performed. The intersection of 3556 DEGs1 and 1622 DEGs2 generated 694 candidates, yielding 7 prognostic genes (UNC13C, MAP7D2, SCARA5 downregulated; SLC35D3, HSPA6, IL31RA, CXCL8 upregulated in tumors). Risk score and pathologic M were independent prognostic factors for nomogram construction, with favorable predictive value. GSEA showed enrichment in pathways like ECM receptor interaction and oxidative phosphorylation; immune correlation, drug sensitivity (e.g., WH.4.023), and RT-qPCR validation of gene expression were confirmed. This study established and verified a novel HDRG-related ESCA prognostic model. The model may serve as a potential tool for risk stratification in ESCA patients and offer new perspectives for exploring the role of histone deacetylation modifications in ESCA progression. However, its clinical translational value, including guiding immunotherapy or targeted drug selection, requires further validation in large-scale prospective studies.

Oral squamous cell carcinoma (OSCC) is a common malignant tumor in the oral and maxillofacial region with a poor prognosis, and its pathophysiology has not been fully elucidated. Although FKBP10 is overexpressed in multiple malignancies, its function and regulatory mechanism in OSCC remain largely unknown. Methods: OSCC and paired adjacent normal tissue samples were collected, and public datasets including The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) were integrated. Bioinformatics approaches including Weighted Gene Co-Expression Network Analysis (WGCNA), limma differential expression analysis and machine learning were applied to screen candidate target genes. Single-cell and spatial transcriptome analyses were performed to delineate the main expression domain of FKBP10 and its distribution in macrophages. Cell Counting Kit-8 (CCK-8), colony formation, Transwell, wound healing and flow cytometry assays were conducted to examine the effects of FKBP10 knockdown on the biological behaviors of SCC-9 cells. Western blot and Gene Set Enrichment Analysis (GSEA) were adopted to explore the potential molecular mechanism. Result: Bioinformatics research identified FKBP10 as a significant target gene for OSCC. FKBP10 is predominantly enriched in fibroblasts and significantly upregulated in OSCC tissues. It has a strong positive correlation with M2-type macrophage infiltration and is linked to a negative outcome for patients. FKBP10 knockdown can cause cell apoptosis, stop the cell cycle at the G0/G1 phase, and prevent SCC-9 cells from proliferating, migrating, and invading. Meanwhile, knockdown of FKBP10 downregulated the protein levels of Periostin (POSTN), Integrin Subunit Beta 5 (ITGB5) and key WNT signaling molecules. Conclusion༚FKBP10 may influence the recruitment of M2-type macrophages by regulating the ECM/integrin adhesion signal and the WNT pathway, thereby changing the tumor immune milieu and encouraging the occurrence and progression of OSCC. It is projected to become a promising biomarker and target for prognostic evaluation and targeted therapy of OSCC.

Copper, an essential trace element with dual functions in cancer progression, drives tumor growth via oncogenic signaling, metabolic plasticity, and extracellular matrix remodeling. By contrast, copper overload triggers cuproptosis, a form of mitochondrial proteotoxic cell death mediated by the FDX1/LIPT1/DLAT/Fe-S regulatory axis. To date, a unified theoretical framework integrating copper metabolism, tumor microenvironment (TME) remodeling, and antitumor immunity remains lacking. In this review, we reframe the TME as a structured copper ecosystem in which both cellular components and the extracellular matrix are modulated by copper, and propose a contextual copper signaling network, in which copper-mediated tumor cell fate is determined by the labile copper pool, the metabolic state, and tumor cellular heterogeneity. We further delineate a unified causal chain linking cuproptosis-driven immunogenicity and cGAS-STING activation to immune cell activation and PD-L1 modulation. Therapeutically, copper chelation and cuproptosis induction strategies have demonstrated promising efficacy, and combining cuproptosis induction with existing antitumor therapies may reverse therapeutic resistance and enhance treatment efficacy. Future studies need to validate cuproptosis-related signatures as predictive biomarkers for precision oncology and refine copper-targeted therapies to minimize systemic toxicities.

<h4>Introduction</h4>Articular cartilage degeneration can lead to a progressive loss of joint function and, eventually, total joint failure. The use of mesenchymal stem cells (MSCs) for cartilage regeneration has proven effective; however, the difficulty of their clinical application has prompted research into other strategies, such as using MSC secretome instead of cells themselves. Our aim was to generate a culture system to obtain large amounts of MSC secretome with a controlled and known composition and the potential to induce articular cartilage regeneration.<h4>Methods</h4>Human dental pulp stem cells (hDPSCs) embedded in alginate/agarose beads were cultured in a bioreactor with constant agitation and periodic culture medium exchange. The secretome was collected and characterized according to known chondral markers (TGF-β1, SERPINE1, miR-140, miR-675, miR-23a, miR-204, miR-211, and miR-337-5p). Metabolomic and proteomic analyses were carried out for detailed characterization of the collected secretome. The chondrogenic potential of the collected secretome was tested in an in vitro model developed using human primary chondrocytes cultured in 3D on alginate-agarose scaffolds.<h4>Results</h4>Our system allowed us to obtain 1200 mL of secretome from intensive cultures of hDPSCs. The cells cultured on the platform remained viable and in an active anabolic state, secreting large amounts of prochondrogenic mediators. Variability between batches of secretome was low, and the collected secretome induced primary chondrocyte differentiation in vitro.<h4>Conclusions</h4>Genomic, metabolomic, and proteomic data indicate that hDPSCs on the platform proliferate and acquire a chondrocyte-like phenotype. These cells secrete mediators that define a microenvironment favorable for chondral regeneration. This is further supported by evidence found in the in vitro differentiation model. Our platform allows the production of large volumes of secretome with controlled composition and chondrogenic induction potential. This is a necessary preclinical study for the subsequent analysis of the secretome obtained using in vivo experimental models.

<h4>Background</h4>Attention-deficit/hyperactivity disorder (ADHD) and language/reading difficulties frequently co-occur. The extent of shared genetic architecture remains incompletely defined. We investigated genome-wide overlap between ADHD and four core skills: word reading, nonword reading, spelling, and phoneme awareness.<h4>Methods</h4>We integrated genome-wide association study (GWAS) summary statistics from the Psychiatric Genomics Consortium for ADHD (38,691 cases; 186,843 controls) and from the GenLang Consortium for language-related traits (<i>n</i> = 13,633-33,959). We performed linkage disequilibrium score regression (LDSC) to estimate genetic correlations and stratified LDSC to identify enriched genomic annotations. Cross-trait meta-analyses were conducted using MTAG and CPASSOC to identify cross-trait association signals. Mendelian randomization (MR) was applied to assess potential directional relationships.<h4>Results</h4>Genome-wide genetic correlations were negative and significant between ADHD and each reading- and language-related trait (word reading rg = -0.35, <i>p</i> = 1.35 × 10^-16; nonword reading rg = -0.28, <i>p</i> = 2.15 × 10^-9; spelling rg = -0.38, <i>p</i> = 1.03 × 10^-15; phoneme awareness rg = -0.28, <i>p</i> = 2.64 × 10^-6). Partitioned heritability analysis using S-LDSC showed significant enrichment after Benjamini-Hochberg false discovery rate correction (BH-FDR), concentrated in conserved or constrained annotations such as Genomic Evolutionary Rate Profiling and phastCons. Cross-trait analyses (MTAG and CPASSOC) identified loci associated with ADHD and each skill, 6 for word reading, 7 for spelling, 7 for phoneme awareness, and 4 suggestive loci for nonword reading (dual-method <i>p</i> < 5 × 10^-6); several map near neurodevelopmental genes including DCC, MEF2C, and ST3GAL3. Bidirectional MR findings were consistent with a potential directional contribution of ADHD genetic liability to poorer reading/language performance, with no clear evidence supporting the reverse direction under standard MR assumptions.<h4>Conclusion</h4>ADHD and language/reading abilities share a substantial polygenic overlap with convergent signals in neuronal regulatory annotations. These genome-wide findings support a transdiagnostic framework linking attention and literacy-related skills, while indicating that causal and clinical interpretations should remain cautious given the modest effect sizes and limited variance captured by current GWAS resources.

Protein succinylation, an emerging post-translational modification (PTM), assumes a crucial role in the initiation and advancement of inflammatory diseases. Ferroptosis, propelled by lethal lipid peroxidation, is intricately associated with the pathogenesis of inflammation. Targeting ferroptosis has recently emerged as a promising therapeutic approach for acute lung injury (ALI). Nevertheless, the crosstalk between protein succinylation and ferroptosis in the regulation of ALI remains ambiguous. FOXO4, a key regulator of oxidative stress responses, is dynamically regulated by various PTMs. We identify SIRT7 as the desuccinylase essential for FOXO4 activation and nuclear localization. Mechanistically, SIRT7 desuccinylates FOXO4 at lysine 139 (K139), thereby inhibiting MDM2-mediated K48-linked polyubiquitination, which stabilizes FOXO4 and maintains FOXO4 nuclear retention. This consequently upregulates glutathione peroxidase 4 (GPX4) and suppresses lipopolysaccharide (LPS)-induced ferroptosis in alveolar epithelial cells (AECs). In vivo experiments demonstrated that SIRT7-knockout (SIRT7-KO) exacerbates LPS-induced ALI. Furthermore, the delivery of a FOXO4-K139R (mimicking desuccinylation) via adeno-associated virus 6 (AAV6) significantly alleviated pulmonary ferroptosis and histopathological damage in SIRT7-KO mice. Notably, pharmacological activation of SIRT7 with trilobatin (TLB) significantly attenuates ALI in LPS-challenged mice, establishing a potential therapeutic pathway for this pathology. Collectively, these findings delineate a previously unrecognized mechanism through which SIRT7 governs the nuclear retention and activation of FOXO4 via desuccinylation, establishing the SIRT7-FOXO4 axis as a potential therapeutic target and theoretical basis for ALI intervention.

Asthma exhibits pronounced circadian variation, yet the molecular mechanisms linking clock disruption to airway epithelial injury remain unclear. In this study, we identify the core clock component BMAL1 as a critical epithelial regulator that restrains ferroptosis-associated injury during allergic airway inflammation. Using a house dust mite (HDM)-induced murine asthma model and HDM-stimulated human bronchial epithelial cells, we found that BMAL1 expression was significantly reduced, whereas BMAL1 deficiency markedly aggravated airway inflammation, mucus metaplasia, and remodeling. Integrated transcriptomic and metabolomic analyses revealed a signature of inflammatory activation and metabolic reprogramming linked to ferroptosis. Consistently, BMAL1 deficiency increased lipid peroxidation and reactive oxygen species levels, downregulated GPX4 and xCT (SLC7A11), and upregulated COX2 expression in the airway epithelium both in vivo and in vitro. Treatment with Ferrostatin-1 attenuated these alterations and partially rescued the aggravated asthmatic phenotype in Bmal1-deficient mice. Notably, constitutive overexpression of BMAL1 also worsened HDM-induced airway pathology, suggesting that disruption of BMAL1 rhythmic oscillation contributes to disease progression. Mechanistically, BMAL1 deficiency activated the AP-1 pathway, induced COX2 expression, and promoted ferroptosis-associated epithelial injury, whereas inhibition of JUN alleviated this phenotype. Furthermore, melatonin also mitigated the aggravated airway pathology and ferroptosis-related changes associated with BMAL1 deficiency. Collectively, these findings identify BMAL1 as a key regulator of airway epithelial ferroptosis and highlight JUN signaling, together with COX2-associated prostaglandin responses, as downstream components of asthma exacerbations driven by circadian dysregulation.

Osteoporosis is a chronic metabolic bone disease that leads to decreased mineral density and causes damage to bone microarchitecture. Elderly persons are more susceptible to bone fractures and have reduced bone strength due to osteoporosis. Conventional pharmacological treatments are the key therapies, but increasing evidence highlights the potential of nutritional interventions to support bone health. Macronutrients, especially proteins from soy, legumes, nuts, seeds, and whole grains, supply essential amino acids vital for type I collagen synthesis, osteoblast differentiation, and bone-forming signalling pathways, promoting bone regeneration and structural integrity. Micronutrients from green leafy vegetables, including magnesium, potassium, vitamin K, vitamin C, zinc, and boron, contribute to mineral homeostasis, enhance calcium absorption, manage oxidative stress, and impact key pathways in bone remodeling, such as RANKL/OPG, Wnt/β-catenin, estrogen modulation, and antioxidant mechanisms. This review examines the current research on the usefulness of macronutrients and micronutrients from numerous plant sources in promoting bone tissue regeneration and reducing dependence on synthetic anti-osteoporotic treatment. It highlights their roles as sustainable, low-risk supplements to traditional osteoporosis treatments by integrating molecular insights with dietary knowledge.

Diabetic kidney disease (DKD) remains one of the most severe microvascular complications of type 2 diabetes mellitus (T2DM) and a leading cause of chronic kidney disease (CKD) worldwide. Nevertheless, despite considerable progress in elucidating its molecular background, early diagnosis and accurate stratification of disease progression remain challenging when relying on conventional clinical biomarkers such as albuminuria and estimated glomerular filtration rate (eGFR). Growing evidence indicates that DKD is driven by interconnected pathogenic mechanisms, including chronic hyperglycemia, activation of the protein kinase C (PKC) signaling pathway, renin-angiotensin-aldosterone system (RAAS) dysregulation, oxidative stress, inflammatory cascades, and immune system activation involving Toll-like receptors (TLR) and the NLRP3 inflammasome. These processes collectively contribute to endothelial dysfunction, podocyte injury, extracellular matrix accumulation, and progressive renal fibrosis. Exosomes and their molecular cargo, particularly miRNAs, have emerged as promising regulators and non-invasive biomarkers reflecting ongoing renal injury. Urinary exosomal microRNAs (uEV-miRNAs) are of interest due to their stability in biological fluids and their direct origin from nephron segments, enabling real-time reflection of renal pathophysiology. Accumulating studies suggest that differentially expressed microRNAs (miRNAs), including miR-21-5p, miR-30a-5p, miR-192-5p, and miR-142-3p, are closely associated with key pathways in DN. However, their clinical translation remains limited by methodological heterogeneity, the lack of standardized isolation protocols, and insufficient validation in large longitudinal cohorts. This review navigates the current landscape of knowledge on the molecular mechanisms underlying DKD and examines the emerging role of uEV-miRNAs as diagnostic biomarkers. Altogether, uEV-miRNAs offer a promising avenue for improving early detection, risk stratification, and disease monitoring in DKD.

Estrogen therapy elicits clinical benefit in ~ 30% of patients with endocrine-resistant estrogen receptor (ER)-positive breast cancer, but its mechanism of action and strategies to increase efficacy remain unclear. Estrogen therapy can induce ER transcriptional hyperactivation and DNA damage; we postulated that such damage could be exacerbated by epigenetic dysregulation via inhibition of histone deacetylases (HDACi). We evaluated the effects of 17b-estradiol and HDACi in three types of ER⁺ breast cancer models: Cells adapted to growth following long-term estrogen deprivation; cells engineered to overexpress exogenous ER that confers endocrine resistance; mice bearing endocrine-resistant patient-derived xenografts. Assay endpoints included apoptosis, growth, DNA damage, histone post-translational modification, levels of ER-regulated transcripts and encoded proteins, cell cycle and replication status, and genome-wide chromatin accessibility, ER binding, and transcriptional profiles. Entinostat treatment increased histone acetylation and chromatin accessibility. Combination treatment with E2 and entinostat inhibited cell growth and induced apoptosis in ER-overexpressing models. E2 and entinostat induced DNA damage as single agents and in combination. These agents synergized against tumor models, offering HDACi as a strategy to enhance efficacy of estrogen therapy.

Aging is accompanied by a gradual loss of cellular function and systemic weakening of the homeostasis of multiple tissues, leading to a variety of age‑related diseases and eventual mortality. Super‑enhancers are a distinct class of cis‑regulatory elements with strong transcriptional activation properties; they are bound by a large number of key transcription factors and cofactors, and possess distinct characteristic modification marks of enhancers. The past decade has seen an exponential growth in the field of super‑enhancer research, partly because of the strong transcriptional activity of super‑enhancers. Super‑enhancers are notably activated in response to aging or aging‑related factors, and serve a pivotal role in aging and aging‑related diseases by modulating key biological processes, including cell proliferation, differentiation and senescence. In the present review, the changes in super‑enhancers under aging‑related stimuli are initially discussed. The mechanistic contributions of super‑enhancers to cellular senescence from the perspective of DNA damage and mitochondrial dysfunction, two hallmarks of aging, are then systematically summarized. Furthermore, the pathogenic involvement of super‑enhancer dysregulation in specific age‑related disorders, including atherosclerosis and type Ⅱ diabetes, is explored, highlighting the potential of super‑enhancers as therapeutic targets. Finally, the current challenges and prospects of super‑enhancers within the context of aging biology are discussed, aiming to provide novel theoretical guidance for the treatment of age‑associated diseases.

<h4>Background</h4>To evaluate the association between tirzepatide use and 1-year risk of major adverse cardiovascular events in patients with type 2 diabetes and atherosclerotic cardiovascular disease, compared with GLP-1 (glucagon-like peptide-1) receptor agonists.<h4>Methods</h4>We conducted a retrospective, propensity score-matched cohort study using the TriNetX network. A total of 16 402 patients with type 2 diabetes and atherosclerotic cardiovascular disease initiating tirzepatide or GLP-1 receptor agonists between January 1, 2022, and March 31, 2025 were matched 1:1. The primary outcome was 1-year major adverse cardiovascular events; secondary outcomes included all-cause mortality, acute myocardial infarction, major adverse limb events, and tissue plasminogen activator use.<h4>Results</h4>Tirzepatide was associated with a lower risk of major adverse cardiovascular events (hazard ratio [HR], 0.75 [95% CI, 0.63-0.91]) and reduced risks of all-cause mortality (HR, 0.69 [95% CI, 0.53-0.90]), major adverse limb events (HR, 0.59 [95% CI, 0.39-0.88]), and acute myocardial infarction (HR, 0.70 [95% CI, 0.53-0.93]), compared with GLP-1 receptor agonists. Results were robust across subgroups and sensitivity analyses.<h4>Conclusions</h4>Among patients with type 2 diabetes and atherosclerotic cardiovascular disease, tirzepatide was associated with a significantly reduced 1-year risk of major adverse cardiovascular events and other cardiovascular outcomes compared with GLP-1 receptor agonists. These findings support the cardiometabolic potential of tirzepatide, warranting further prospective validation.

<h4>Purpose</h4>Trastuzumab deruxtecan (T-DXd) has improved outcomes in metastatic breast cancer; however, a substantial subset of patients experience early lack of clinical benefit that is not reliably predicted by routine clinicopathologic variables. In an institutional cohort of 109 T-DXd-treated metastatic lesions, HER2 expression level, ER/PR status, Ki-67 index, and metastatic site were not significantly associated with response, highlighting the need to define mechanistic determinants of intrinsic resistance.<h4>Experimental design</h4>We performed spatial transcriptomic and proteomic profiling using NanoString GeoMx Digital Spatial Profiling on pretreatment bone, brain, and soft-tissue metastases from patients with clinical benefit (response) versus early progression (resistance) on T-DXd. Tumor (PanCK⁺) and immune (CD45⁺) compartments were analyzed to link tumor architecture and region-resolved signaling states with therapeutic response. Candidate resistance pathways were functionally evaluated in HER2-positive breast cancer cell lines and in vivo metastasis models treated with T-DXd alone or combined with the RAGE inhibitor TTP488.<h4>Results</h4>Spatial proteogenomics revealed recurrent upregulation of S100 family alarmins in resistant tumor regions, associated with activation of a RAGE-centered pro-survival signaling program characterized by ERK, AKT, and STAT3 phosphorylation. Pharmacologic RAGE inhibition enhanced T-DXd-induced apoptosis, restored drug sensitivity in vitro, and significantly reduced metastatic burden in lung and brain metastasis models.<h4>Conclusions</h4>Spatial proteogenomics identifies a conserved S100-RAGE-driven survival state coupled to immune-excluded tumor architecture as a mechanism of intrinsic T-DXd resistance across metastatic niches. Targeting this pathway with an orally available RAGE antagonist restores T-DXd responsiveness and offers an immediately translatable strategy to overcome resistance in metastatic breast cancer.

The RNA-binding protein Sex-lethal (Sxl) is classically known as a master regulator of sex determination and mRNA splicing in Drosophila melanogaster. However, this role is not conserved across species, and functions beyond the canonical pathway remain poorly understood. In this study, we uncover a splicing-independent role for Sxl at the chromatin level in the Drosophila brain. Using Targeted DamID (TaDa) profiling in neurons, we identify widespread binding of Sxl to promoter regions, independent of sex or RNA binding activity. Notably, Sxl chromatin occupancy exhibits near-complete overlap with Polr3E (RPC37), an RNA Polymerase III subunit, with Sxl binding abolished upon Polr3E knockdown. Depletion of Sxl in mature male neurons induces widespread transcriptional changes, particularly in metabolic genes, and improves negative geotaxis during aging, phenotypes that closely mirror Polr3E knockdown. Conversely, overexpression of the brain-specific SxlRAC transcript leads to severe climbing deficits and upregulated gene expression associated with metabolism and translation. Manipulating Sxl levels in the brain significantly impacts select tRNA production and global protein synthesis rates. Together, these findings reveal a previously unrecognized role for Sxl in regulating Pol III activity via Polr3E, modulating tRNA synthesis and supporting neuronal metabolism. Given the emerging tie between Pol III regulation and neuronal aging, our study highlights Sxl as a novel factor in neuronal homeostasis.

Neuroblastoma, a pediatric cancer derived from sympathetic ganglia of the peripheral nervous system, frequently metastasizes, driving poor outcomes. Primary neuroblastomas are well-characterized, but the mechanisms underlying metastasis remain poorly understood. Here, by using single-cell and spatial multiomics, we identified that lymph node metastases in high-risk neuroblastomas display distinctive cellular heterogeneity and plasticity, marked by mesenchymal-like and stem-like states and heightened epithelial-to-mesenchymal transition activity compared to primary adrenal tumors. In addition, compared to primary adrenal masses, the metastatic niche display increased immunosuppressive myeloid programs, heightened immune checkpoint signaling, and lymphocyte exhaustion, which are indicative of immune evasion and dysfunction. Notably, metastatic neuroblastomas show elevated eIF4F translation machinery and XPO1 levels. Dual inhibition of eIF4A and XPO1 synergistically halted tumor growth and prolonged survival in xenograft models. Together, our multiomics studies reveal the molecular and cellular plasticity that contributes to therapy resistance and highlight exploitable therapeutic vulnerabilities in high-risk metastatic neuroblastomas.

Accurate and early detection of pneumonia is crucial for effective treatment; however, current diagnostic methods often lack the necessary specificity and sensitivity. Herein, we begin with a comprehensive bioinformatics analysis, identifying neutrophil elastase (NE) as a critical biomarker associated with pneumonia progression. We subsequently develop an NE-responsive probe (NERP), composed of a hemicyanine fluorophore linked to an NE-sensitive peptide, specifically designed for activation in inflamed tissues. To facilitate urinalysis, NERP is further refined into a hydrophilic active targeting responsive probe (ATRP_H). ATRP_H demonstrates exceptional sensitivity in both in vivo imaging and urine-based detection, with urine analysis offering a noninvasive, early-stage diagnostic option that overcomes the limitations of tissue penetration and low accuracy. In addition, ATRP_H is highly effective in drug screening, dosage optimization, and exploring mechanisms of NE production. This biomarker screening and design strategy not only enhances pneumonia diagnosis and treatment via different routes of administration but also has broader potential for other inflammatory lung diseases.

Enhancers are abundant and critical gene-distal cis-regulatory elements with distinct architectural features; however, a mechanistic understanding of their interactions within endogenous chromatin contexts remains challenging. Here, we developed a recombinase-mediated genome-rewriting platform to explore how a long-range human enhancer, eNMU, confers a remarkable 10,000-fold activation of its target gene, Neuromedin U (NMU), at its native locus. Our systematic dissection reveals two functionally distinct sub-elements of eNMU: the canonical autonomous enhancer e1 and the intrinsically inactive facilitator e2, which dramatically augments e1's activity. The autonomous enhancer e1 is functionally hierarchical to e2, additional facilitators, and the NMU promoter across the ∼100-kb NMU-eNMU region, and it orchestrates the formation of a 3D regulatory hub. e1 also harbors a bipartite structure: a divergently transcribed retroviral long terminal repeat (LTR) enhancer and an adjacent LTR promoter that dampens NMU expression. We explore and discuss the broader implications of our focused study for understanding enhancer regulatory mechanisms genome-wide.

Toll-like receptors (TLRs) are considered general sensors of bacterial encounters. Here, we examined whether other pattern recognition receptors are commonly activated during bacterial infection. TLR-independent interferon (IFN) responses were induced in macrophages in response to diverse bacterial encounters. Of the cytoplasmic receptor families examined, the cyclic dinucleotide (CDN) sensor STING was required for IFN responses to evolutionarily diverse bacteria. Various bacterial CDNs were present in murine tissues; these activated stimulator of interferon genes (STING) after bacteriolysis in phagolysosomes in a manner requiring two CDN transporters. Importantly, bacterial CDNs were increased in colonic biopsies from patients with inflammatory bowel disease. Systemic delivery of dead, CDN-laden bacteria promoted anti-tumor immunity in mice. Detection of diverse CDNs, including pyrimidine-based CDNs, was an evolutionarily conserved feature of STING, with distinct binding modes for purine- and pyrimidine-based CDNs. Thus, a phagocytosis-CDN-STING connection places cytoplasmic sensing as a common outcome of host-bacteria interactions that set the immune tone of a tissue, with implications for host defense.

The endoplasmic reticulum (ER), a major calcium ion (Ca²⁺) reservoir, plays a pivotal role in lipid synthesis, protein synthesis, and secretion. Disruption of ER function leads to the accumulation of misfolded proteins, resulting in ER stress. To restore cellular homeostasis, the unfolded protein response (UPR) is activated. However, prolonged or irreversible ER stress can trigger apoptosis and cell death. Cellular dysfunction and apoptosis arising from disrupted ER Ca²⁺ homeostasis are often implicated in neurodegenerative and metabolic disorders. We investigated the role of LRRC8B, an ER-resident protein previously linked to ER Ca²⁺ homeostasis, in the ER stress response. LRRC8B expression was upregulated in response to chemically induced ER stress. Overexpression of LRRC8B enhanced the viability of HEK293T cells exposed to the ER stressor tunicamycin, whereas LRRC8B knockdown increased their susceptibility to apoptosis. Furthermore, LRRC8B overexpression reduced protein aggregation during ER stress by upregulating cytoprotective genes associated with the adaptive UPR, including BiP, calnexin, and PDI. Conversely, LRRC8B knockdown decreased BiP expression and elevated the levels of ER stress-induced apoptotic proteins such as CHOP, Bax, and cleaved caspase-3. Co-treatment with the chemical chaperone, 4-PBA, partially rescued the LRRC8B knockdown cells undergoing apoptosis. In Neuro 2a (N2a) cells, LRRC8B overexpression diminished the aggregation of mutant huntingtin protein (HTT-83Q), associated with Huntington's disease, whereas its knockdown exacerbated HTT-83Q aggregation. Taken together, these findings suggest that LRRC8B is a key modulator of the ER stress response and plays a cytoprotective role.

<h4>Objectives</h4>SAMM50 rs3761472 is associated with metabolic dysfunction-associated steatotic liver disease (MASLD), but its functional consequences in vivo remain unclear. We investigated whether this variant disrupts mitochondrial function and promotes MASLD progression.<h4>Methods</h4>Associations of rs3761472 with MASLD and liver-related traits were evaluated using Korea Biobank Array data. We generated Samm50 knock-in (KI) mice carrying the D110G substitution corresponding to human rs3761472 using CRISPR/Cas9 and assessed hepatic mitochondrial homeostasis and MASLD-related phenotypes in mice fed a normal diet or a high-fat diet.<h4>Results</h4>In human genetic analyses, rs3761472 was significantly associated with MASLD and higher serum levels of liver injury markers. Samm50-KI mice showed reduced hepatic SAMM50 expression, disrupted mitochondrial organization, impaired mitochondrial respiration and ATP production, increased mitochondrial oxidative stress, inflammatory activation, apoptosis, and liver injury. Following high-fat diet feeding, Samm50-KI mice exhibited greater hepatic lipid accumulation and liver injury, together with more pronounced insulin resistance and glucose intolerance, than wild-type mice.<h4>Conclusions</h4>Our findings establish rs3761472 as a functional genetic variant linking mitochondrial architecture to metabolic liver disease pathogenesis, with potential relevance as a genetic biomarker for MASLD susceptibility.

<h4>Objective</h4>To develop and validate a non-invasive model for predicting early recurrence (ER) after microwave ablation (MWA) in patients with hepatocellular carcinoma (HCC) and comorbid type 2 diabetes (T2D).<h4>Methods</h4>This retrospective study enrolled 186 HCC patients with T2D who underwent MWA and were divided into training (n=149) and validation (n=37) cohorts. Preoperative clinical parameters and portal-venous phase MRI radiomic features were extracted. A radiomics score (RAD -score) was constructed using the K-nearest neighbors (KNN) algorithm after feature selection. A clinical model was built using logistic regression, and a nomogram integrating clinical factors and the RAD -score was established. Model performance was evaluated using ROC curves, calibration curves, and decision curve analysis (DCA). Kaplan--Meier curves, pathological analysis, and immunohistochemistry validated prognostic stratification.<h4>Results</h4>Four core radiomic features were identified. The combined nomogram achieved AUCs of 0.877 (training) and 0.846 (validation), outperforming the clinical (AUC 0.718) and radiomics (AUC 0.774) models alone. In the validation cohort, the sensitivity, specificity, and negative predictive value were 80.0%, 83.8%, and 90.1%, respectively. DCA showed a significant net benefit across threshold probabilities of 0.1-0.9. High-risk patients had a 5.8-fold (training) and 2.51-fold (validation) higher recurrence risk with significantly shorter progression-free survival. Pathological and immunohistochemical analyses confirmed more aggressive tumor features (higher rates of microvascular invasion, poor differentiation, thick-trabecular type, CD34 expression, and glypican-3 expression) in the high-risk group.<h4>Conclusion</h4>The nomogram provides a satisfactory prediction of early recurrence within two years after MWA in HCC patients with T2D, enabling individualized postoperative surveillance and intervention.

<b>Background/Objectives</b>: Long non-coding RNAs (lncRNAs) are important regulators of tumor biology through their interactions with DNA, proteins, and non-coding RNAs. Although ENST00000615487.1 (also known as CTD-2396E7.11/AC010503.4) has been associated with multiple malignancies, its biological role in colorectal cancer (CRC) remains poorly characterized. This study aimed to investigate the expression pattern, cellular and subcellular localization, and potential functional role of ENST00000615487.1 in CRC using integrated in vitro and in silico approaches. <b>Methods</b>: Molecular characteristics of the transcript were obtained with the CPC2 and RNA Analyzer 3 tools. Differential expression of ENST00000615487.1 across 10 tumor types was analyzed using the UCSC Xena Browser. Transcript expression was experimentally evaluated in normal, tumor, and fibroblastic colon cell lines by PCR, while subcellular localization was assessed through the lncATLAS, lncLocator, and iLoc-LncRNA tools, and experimentally confirmed by qRT-PCR. Single-cell RNA sequencing data from the GSE161277 dataset were analyzed to determine cell type-specific expression patterns. Potential interactions with DNA, miRNAs, and proteins were investigated using Fasim-LongTarget, miRDB, and AnnoLnc2, followed by functional enrichment analyses using STRING and Enrichr. <b>Results</b>: ENST00000615487.1 was identified as a structurally stable non-coding transcript with a highly organized secondary structure. Differential expression analysis demonstrated significant downregulation in CRC compared with that in normal colon tissue. Single-cell transcriptomic analysis revealed predominantly epithelial-specific expression. In silico and experimental analyses demonstrated predominant nuclear localization in normal colon cells, whereas cytoplasmic enrichment was observed in CRC cells. Functional analyses identified potential interactions with <i>HIP1R</i>, <i>RPH3AL</i>, specific miRNAs, and proteins involved in transcriptional regulation and RNA processing pathways, as well as functional connections with proteins involved in vesicular transport. <b>Conclusions</b>: ENST00000615487.1 is a structurally stable lncRNA exhibiting context-dependent expression and localization patterns in CRC, suggesting a potential shift from nuclear transcriptional regulation toward cytoplasmic post-transcriptional functions during colorectal carcinogenesis.

The ZFP36 family proteins (TTP, ZFP36L1, and ZFP36L2) are RNA-binding proteins that function as key post-transcriptional regulators of gene expression. They bind AU-rich elements (AREs) in target mRNA 3'UTRs, recruit the CCR4-NOT deadenylation complex to trigger mRNA decay, and maintain homeostasis in immunity, barrier function, and stem cell fate. Rather than acting on single targets, all family members share a conserved mRNA destabilization mechanism, with outcomes determined by member-specific expression, kinase-mediated regulation, and cell-type-dependent target availability. Dysregulation of this network stabilizes mRNAs encoding pro-inflammatory cytokines, immune checkpoints, and oncogenes, driving pathogenesis of inflammation, autoimmunity, cancer, cardiovascular and neurodegenerative diseases. Individual family members exert context-dependent and sometimes opposing effects, so their net function depends on the specific cellular and disease context. Therapeutic strategies targeting ZFP36 activity, including phosphatase agonism and epigenetic modulation, have shown promising preclinical results, but clinical translation remains early. This review summarizes the molecular regulatory networks of the ZFP36 family and their physiological and pathological roles, emphasizing the mechanistic principles that unify family-member function and the contextual factors that diversify it, to provide a foundation for future therapeutic development.

Pancreatic acinar cell carcinoma (PACC) is a rare exocrine pancreatic malignancy accounting for approximately 1-2% of adult pancreatic neoplasms. Although historically grouped with pancreatic ductal adenocarcinoma (PDAC), accumulating evidence demonstrates that PACC represents a biologically distinct entity with unique clinical, pathologic, and molecular characteristics. Compared with PDAC, PACC more commonly presents as a large pancreatic mass, is less frequently associated with obstructive jaundice, and exhibits lower rates of <i>KRAS</i> mutations. Recent genomic studies have identified recurrent alterations involving DNA damage repair pathways, WNT/β-catenin signaling, and actionable kinase fusions, including <i>BRAF</i> and <i>RAF1</i> rearrangements. In advanced disease, fluoropyrimidine- and platinum-based regimens appear to demonstrate greater activity than traditional gemcitabine-based approaches, although prospective comparative data are lacking. This review summarizes the current understanding of the epidemiology, clinical presentation, pathology, molecular landscape, treatment approaches, and prognostic factors associated with PACC. We highlight emerging opportunities for precision oncology and discuss ongoing challenges in the management of this uncommon pancreatic malignancy.

Asthma is a highly heterogeneous chronic inflammatory disease of the airways, among which Th2-high asthma represents the most prevalent endotype. The pathogenesis of Type 2 (T2) asthma (driven by type 2 inflammation) involves a complex immune network orchestrated by the coordinated actions of multiple effector cell populations, including Th2 cells, group 2 innate lymphoid cells (ILC2s), and type 2 cytotoxic T (Tc2) cells. Epithelial-derived alarmins, particularly interleukin-33 (IL-33) and thymic stromal lymphopoietin (TSLP), function as key upstream initiators that bridge innate and adaptive immunity. In addition, multilayered regulatory mechanisms-including genetic susceptibility, metabolic reprogramming, and ubiquitination-collectively govern the initiation and progression of Th2-driven inflammation. With the deepening understanding of these mechanisms, therapeutic strategies have progressively shifted from targeting downstream effector molecules to upstream alarmins, thereby providing new directions for precision medicine. This review systematically summarizes recent advances in the immunopathogenesis and targeted therapies of T2 asthma, offering a conceptual framework for precision-based clinical interventions.

<h4>Introduction</h4>In recent years, the clinical relevance of tertiary lymphoid structures (TLS) in cancer has become increasingly clear. However, the mechanisms that promote TLS development have remained obscure, largely because of a lack of animal models in which cause and effect studies can be performed. In this study, we used a mouse model in which TLS develop spontaneously in intraperitoneal tumors to address this issue.<h4>Methods</h4>Cxcl13-cre-tdTomato mice were implanted with B16-OVA melanoma tumors in either intraperitoneal or subcutaneous locations to establish which cells expressed CXCL13. Single cell suspensions from tumors were analyzed by flow cytometry. To address the impact of CXCL13⁺ CAF on TLS formation, we bred Cxcl13-cre tdTomato mice to R26R-iDTR^flox mice, which express the diphtheria toxin receptor under the control of a floxed stop codon. The resulting Cxcl13-cre/iDTR mice were treated with diphtheria toxin beginning on day 7 after IP tumor implantation. Single cell suspensions from tumors were analyzed by flow cytometry. To characterize their functional properties, CAF in IP melanoma tumors from Cxcl13-cre/eGFP mice were collected by cell sorting, analyzed for transcript expression using scRNA-seq, clustered into groups using the Seurat package, and profiled using pathway analysis and differential gene analysis.<h4>Results</h4>Extending earlier studies in which we showed that CXCR5-expressing B cells are essential for TLS development, we found that CXCL13, the ligand for CXCR5, were expressed exclusively by a population of cancer-associated fibroblasts (CAF), and that these CAF were essential to promote B cell accumulation in these tumors. In keeping with earlier work showing that the presence of TLS was associated with smaller tumor size, we also found that the lack of B cell accumulation was associated with larger tumor size. Using scRNA-seq, we identified 7 groups of CAF in TLS containing tumors. One of these CAF groups was substantially enriched for CXCL13-expressing cells. This population was also enriched for additional genes that could promote B cell recruitment and enable regulation by a variety of cytokines. However, other genes that we have shown or hypothesize to be important in TLS development were enriched for expression in other CAF groups.<h4>Conclusions</h4>This work indicates that TLS development in this model depends on multiple CAF populations operating at distinct points in the developmental process. Some of these populations appear similar to, but distinct from, those defined in other human and murine tumor models. However, we also discriminated new immunologically relevant populations of CAF, particularly one expressing CXCL13, which we propose acts as a central organizer of tumor-associated TLS.

<h4>Background</h4>Small cell lung cancer (SCLC) is a highly aggressive malignancy characterized by rapid progression, early metastasis, frequent relapse, and treatment resistance. Although chemoimmunotherapy has improved outcomes in a subset of patients, reliable molecular stratification tools reflecting tumor heterogeneity and survival risk remain limited. Chaperone-mediated autophagy (CMA) is involved in proteostasis, metabolic adaptation, and stress responses, but its cellular heterogeneity and prognostic relevance in SCLC remain unclear.<h4>Methods</h4>This study integrated single-cell RNA sequencing data, bulk transcriptomic cohorts, and clinical information to characterize CMA-related heterogeneity in SCLC. At the single-cell level, SCLC subtypes were annotated, CMA scores were calculated, and differences among baseline, sensitive, and resistant states were evaluated. At the bulk level, tumor versus normal differential expression analysis and weighted gene co-expression network analysis were performed to identify CMA-related candidate genes. A CMA-related Risk Score was constructed through comprehensive machine learning comparison and evaluated using survival analysis, time-dependent receiver operating characteristic curves, Cox regression, nomogram analysis, functional enrichment, immune microenvironment analysis, and drug sensitivity prediction. ARRDC3 was further validated by <i>in vitro</i> experiments.<h4>Results</h4>Single-cell analysis revealed marked SCLC subtype heterogeneity and nonuniform CMA activity across cellular subtypes. Significant CMA score differences between sensitive and resistant cells were observed in the SCLC-A_NR0B1/MYCL+ and SCLC_Hypoxia_glycolytic subtypes. Integration of CMA-related co-expression modules with tumor-associated differentially expressed genes identified 53 candidate genes. The machine learning-derived Risk Score effectively stratified patients into high-risk and low-risk groups across the combined cohort, GSE60052 cohort, and cBioPortal cohort, with high-risk patients showing significantly poorer overall survival. The Risk Score remained an independent prognostic factor and was associated with proliferative, cell-cycle, metabolic, immune, and drug sensitivity-related features. <i>In vitro</i> experiments showed that ARRDC3 overexpression suppressed proliferation, colony formation, migration, and invasion in H446 cells.<h4>Conclusion</h4>This study revealed CMA-related cellular heterogeneity in SCLC and established a robust prognostic Risk Score associated with survival, biological pathway activity, immune microenvironment features, and potential therapeutic responses. Functional validation of ARRDC3 further supports the biological relevance of this model, providing new insights into SCLC molecular risk stratification and CMA-associated resistance states.

Despite therapeutic advances, multiple myeloma (MM) remains characterized by progression and relapse, underscoring the need for reliable biomarkers for early detection and monitoring. Extracellular vesicles (EV) are cell-derived, lipid bilayer-enclosed particles that mediate intercellular communication and circulate in body fluids, making them promising biomarkers. This study compares the proteome of MM-derived EV with parental cells and evaluates whether EV represent a superior source of MM-specific biomarkers. We used high-resolution mass spectrometry to analyze MM cell lines and their EV. Cell line-derived EV offer a cleaner, tumor-specific protein profile, minimizing background from nonmalignant cells and improving biomarker accuracy. MS datasets were subjected to bioinformatic analysis, followed by functional enrichment using gene set libraries such as Gene Ontology, Jensen Compartments, TRRUST and DisGeNET, to extend interpretation beyond limited protein databases. Compared to cellular proteins, the EV proteome was enriched in functions related to oncogenic signaling, transcriptional regulation and MM pathophysiology, strengthening its potential as a cancer-related biomarker source. DisGeNET identified cancer-related proteins that were further evaluated to identify MM-specific candidates analyzing MM patients' gene expression databases containing clinical data. This led to the identification of macrophage migration inhibitory factor (MIF) and prohibitin. ELISA on plasma from MM patients detected significantly increased MIF levels in smoldering and active MM compared with healthy donors. Approximately half of circulating MIF was EV-associated in MM patients, indicating that MIF is distributed between vesicular and soluble plasma fractions and underscoring its potential as a noninvasive biomarker for MM early detection and patient monitoring.

Rheumatic heart disease (RHD) remains the leading cause of acquired valvular heart disease in low- and middle-income countries, affecting an estimated 40 million individuals and disproportionately affecting children and young adults in endemic regions. No pharmacological intervention has been shown to modify valvular disease progression in established RHD. Sodium-glucose cotransporter-2 (SGLT2) inhibitors have demonstrated cardioprotective effects across multiple cardiovascular conditions, prompting interest in their potential applicability to RHD. This hypothesis-generating review synthesizes current evidence on the molecular and cellular pathology of RHD and evaluates the mechanistic plausibility of SGLT2 inhibitor effects in this context. Evidence derived largely from non-rheumatic valve disease and experimental models suggests that SGLT2 inhibitors may attenuate oxidative stress, inflammatory cytokine signaling, transforming growth factor-β-driven fibrosis, and mitochondrial dysfunction; processes implicated in valvular remodeling. Mechanotransduction cascades and osteogenic calcification pathways, characterized primarily in calcific aortic valve disease, are biologically plausible but remain uncharacterized in RHD tissue. Whether these mechanisms are operative within the postinflammatory, immune-mediated rheumatic valve environment remains uncertain and represents an important avenue for future research. Validation of SGLT2 expression in rheumatic valve tissue, mechanistic testing in etiologically relevant models, and early-phase clinical trials tailored to endemic health system realities are essential prerequisites before any therapeutic claims can be advanced. This review outlines the translational pathway needed to determine whether this hypothesis warrants clinical development.

Sickle cell disease and cancer represent fundamentally distinct classes of human disease-one is driven by a defined mutation in β-globin, whereas the other arises through complex genetic and epigenetic alterations that reshape cellular identity and behavior. Despite these differences, both contexts illustrate how transcription factors, chromatin regulators, and cis-regulatory elements can impose disease-relevant gene expression states. In β-hemoglobinopathies, therapeutic reactivation of fetal hemoglobin through modulation of γ-globin (HBG1/2) regulatory pathways, most notably disruption of the erythroid-specific BCL11A enhancer, has emerged as a clinically validated strategy. These advances have been facilitated in part by the HUDEP-2 erythroid progenitor cell line, which provides a tractable adult erythroid model for identifying fetal hemoglobin regulators, validating their function, and evaluating relevant gene editing and gene-regulatory therapies. Many of the regulators implicated in γ-globin silencing, including BCL11A, ZBTB7A, NuRD-associated proteins, DNMT1, KDM1A/LSD1, MYB, and ATF4, also function in cancer-associated transcriptional or epigenetic networks. In cancer, these factors can support oncogenic transcription, tumor suppressor repression, impaired differentiation, stress adaptation, invasion, or therapy resistance. This review summarizes discoveries enabled by HUDEP-2 cells in fetal hemoglobin regulation and hemoglobinopathy therapeutic development, then discusses how these mechanisms provide conceptual parallels for understanding and targeting regulatory dependencies in cancer.

<h4>Background & aims</h4>Hirschsprung-associated enterocolitis, a lethal complication of Hirschsprung's disease-a prevalent congenital disorder with high morbidity-has a poorly understood pathogenesis. We investigated how aberrant immune-stromal crosstalk drives mucosal injury in Hirschsprung's disease.<h4>Methods</h4>Integrated single-cell RNA sequencing, spatial transcriptomics, bulk RNA sequencing, and proteomics were performed using human Hirschsprung's disease tissues. Validation was conducted using murine Hirschsprung-associated enterocolitis (Ednrb^-/-) models, flow cytometry, multiplex immunohistochemistry, in vitro B-cell stimulation, epithelial coculture, rescue experiments with the thymic stromal lymphopoietin inhibitor baicalein, and ex vivo intestinal organoid assays.<h4>Results</h4>We identified significant expansion and hyperactivation of B cells that colocalized with epithelial barrier defects in Hirschsprung's disease segments. Multiomics analysis revealed an activated phenotype in these naive and memory B cells, characterized by the upregulation of B lymphocyte kinase, B cell linker, and proliferating cell nuclear antigen, which may contribute to impaired epithelial homeostasis. Crucially, we identified profibrotic thymic stromal lymphopoietin+ fibroblasts enriched in Hirschsprung's disease. Spatial mapping and in vitro assays demonstrated that thymic stromal lymphopoietin potently activated B cells, synergizing with classical stimuli to enhance proliferation. Activated B cells, in turn, secreted proinflammatory factors, such as, ICAM1 and CCL5, which induced epithelial apoptosis, suppressed proliferation, and disrupted tight junctions-effects that were amplified by thymic stromal lymphopoietin costimulation. Activated B cells and epithelial dysfunction were conserved in Hirschsprung-associated enterocolitis (Ednrb^-/-) mouse models. Coculture and ex vivo organoid experiments confirmed that thymic stromal lymphopoietin-activated B cells directly destabilized epithelial homeostasis; crucially, baicalein treatment rescued epithelial proliferation, attenuated apoptosis, and reduced inflammation-associated gene expression.<h4>Conclusions</h4>Our study reveals a novel pathogenic mechanism whereby thymic stromal lymphopoietin instigates pathogenic B-cell hyperactivation, thereby disrupting epithelial integrity and increasing susceptibility to Hirschsprung-associated enterocolitis. Inhibition of thymic stromal lymphopoietin signaling with baicalein ameliorates epithelial injury, highlighting this axis as a promising therapeutic target for Hirschsprung-associated enterocolitis prevention in patients with Hirschsprung's disease.

Risdiplam and branaplam represent two classes of small-molecule splicing modulators that promote U1 snRNP recognition of weak non-canonical GA/GU-containing 5' splice sites (ss). We demonstrate that branaplam enhances recognition of these 5' ss by reconstituted U1 snRNP in vitro, and that this effect depends on the ZnF domain of U1C and Helix H of U1 snRNA, but not U1A or U1-70K. We also demonstrate that branaplam enhances the weak 5' ss recognition through a dual act of strengthening the U1 snRNP-5' ss interaction and U1 snRNP-U1C interaction. In cells, depletion of U1C reduces or abolishes compound-induced exon inclusion for most cassette exons. Interestingly, a subset of cassette exons become responsive to compound only upon U1C knockdown, supporting a model in which U1C stabilizes specific conformations at the 5' ss-U1 snRNA interface in a context-dependent manner that can either facilitate or hinder compound binding. Surprisingly, risdiplam shows no effect on weak 5' ss recognition in vitro, suggesting additional cellular factors are required for its activity.

<h4>Background</h4>Heat stress (HS) poses a major challenge to the dairy industry by reducing milk production, yet its cell type-specific effects in the bovine mammary gland remain incompletely defined. In this study, we recorded production traits and collected mammary biopsies from cows under thermoneutral (TN), HS, and pair-fed (PF) conditions.<h4>Results</h4>Clinical measurements confirmed HS-induced physiological alterations. Compared with TN cows, HS cows exhibited reduced dry matter intake (DMI), milk yield, and yields of fat, protein, and lactose, along with increased water intake and milk urea nitrogen. The use of PF controls indicated that decreased DMI accounted for 45% of the milk-yield reduction, whereas direct HS effects accounted for the remaining reduction. We applied single-nucleus RNA-seq (snRNA-seq) on mammary biopsies to generate cell-resolved HS responses. We identified 14 distinct cell clusters, including epithelial, immune, and stromal populations. Under the TN condition, casein genes (e.g., CSN1S1, CSN2) were broadly expressed across luminal cells but were attenuated under HS, whereas luminal alveolar cells showed relative upregulation. Heat shock protein genes were strongly induced by HS, primarily in epithelial clusters. Gene-set enrichment analyses revealed increased ribosomal activities across HS-responsive clusters and enrichment of protein folding and metabolic pathways in luminal alveolar cells, suggesting elevated proteostasis demands under stress. Pseudotime analysis positioned luminal cells along a progenitor-to-secretory trajectory under TN, accompanied by increased casein gene expression, whereas under HS, mature luminal cells shifted toward a homeostasis regulatory state. Cell-cell communication analysis demonstrated HS-induced remodeling of interepithelial signaling, including altered ERBB4-mediated signaling from luminal hormone-sensing to alveolar lineages. Finally, transcription factor activity profiling highlighted cell type-specific HS-activated regulators and their downstream target genes.<h4>Conclusions</h4>Together, this cell type-resolved atlas delineates how HS alters bovine mammary epithelial function, developmental state, and intercellular crosstalk. These findings point to proteostasis pressure, disrupted signaling pathways, and rewired regulatory networks as mechanistic contributors to reduced lactational performance under HS, offering insights for improving heat resilience in dairy cattle.

Subplate neurons (SpNs) are among the earliest-born and maturing neurons in the developing cerebral cortex. They arise from multiple origins and can be classified into several subgroups based on morphology, connectivity, and gene expression. These neurons play essential roles in cortical circuit formation, yet their cellular diversity and transcriptional dynamics remain incompletely understood. Here, we characterized transcriptomic profiles of SpN subpopulations in embryonic mouse cortex using Lpar1-EGFP and NeuroD1/Cre-ERT2 (D1B) reporter lines. We applied complementary approaches of gene expression profiling, including bulk microarray analysis, single-cell RNA sequencing (scRNA-seq), and Visium spatial transcriptomics. At embryonic day 17 (E17), scRNA-seq identified 10 distinct Lpar1-EGFP-positive SpN clusters, which spatial transcriptomics mapped to specific cortical regions. While many markers showed enrichment within the subplate region, others extended into the hippocampus and ventral pallium (including the amygdala, claustrum, and endopiriform nucleus). Integrated analysis of Lpar1-EGFP and D1B lines revealed both overlapping and unique gene expression signatures, highlighting dynamic markers of subplate identity. Comparisons between E15 and E17 datasets showed substantial transcriptional shifts, suggesting rapid developmental changes in SpN subgroups. Validation with in situ hybridization and RNAscope confirmed the selectivity of key markers, including Cryab, Cdh13, Nr4a2, and Lmo3. Together, these findings provide a molecular framework for further classifying SpN subtypes and identifying candidate markers for transient versus persistent populations, thereby advancing our understanding of early cortical development.

Antipsychotic-associated metabolic toxicity remains one of the most persistent clinical problems in psychopharmacology. Clozapine and olanzapine are especially effective for psychosis but carry high liability for weight gain, dyslipidemia, insulin resistance, and type 2 diabetes. Current monitoring recommendations recognize this risk, yet they remain largely uniform across patients and do not incorporate ancestry-specific genetic risk or mechanistic drug-gene information. We performed a transcriptome-wide association study-informed drug-gene prioritization analysis to examine whether approved antipsychotic target genes overlap with genes whose genetically predicted expression is associated with type 2 diabetes. The analysis used ancestry-specific type 2 diabetes transcriptome-wide association study (TWAS) results derived from a multi-ancestry genome-wide association study (GWAS) and approved antipsychotic drug-gene interactions from the Drug-Gene Interaction Database (DGIdb). For each drug, target genes were matched to TWAS genes across six metabolic tissues, and a weighted risk score was calculated as the sum of the absolute TWAS z-score multiplied by the drug-gene interaction score for significant targets. Follow-up analyses decomposed signals into targets with positive and negative TWAS directions, operationally interpreted as aggravating and compensatory, while also examining curated metabolic axis genes including GLP1R, GIPR, PPARG, and SLC2A4. The analysis identified recurrent exploratory target-overlap signals for clozapine and olanzapine. Clozapine showed the most consistent cross-ancestry aggravating profile, with recurrent target overlap involving GLP1R and immune-related genes. Olanzapine showed strong mechanistic-axis overlap involving GLP1R, GIPR, and PPARG, although its simple TWAS directionality was often classified as compensatory. Trifluoperazine emerged as a notable candidate, with significant target enrichment in the European ancestry analysis and top ranking in the Hispanic analysis. Fluspirilene also met the combined enrichment false discovery rate threshold in the European ancestry analysis, although its clinical metabolic interpretation was less direct. Haloperidol decanoate showed a high burden driven partly by SLC2A4, but its directionality was frequently mixed or compensatory. These findings nominate the incretin axis as a plausible translational bridge between antipsychotic metabolic liability and existing interventions such as GLP-1 receptor agonists. They also identify a key methodological gap. Future models must incorporate the pharmacologic mode of action to distinguish receptor blockade from activation. Except for the enrichment-positive European ancestry findings for trifluoperazine and fluspirilene, the results are exploratory prioritization signals and do not establish drug-specific metabolic effects, causal mechanisms, or ancestry-specific treatment effects. Overall, this TWAS-informed analysis provides a hypothesis-generating framework for ancestry-aware metabolic monitoring and targeted validation studies.

FMS-like tyrosine kinase 3 (FLT3) is a key driver of acute myeloid leukemia (AML); mutations within FLT3, specifically ITD lesions and TKD point mutations, promote proliferation and are associated with poor prognosis. Although FLT3 inhibition is central to AML therapy, resistance, particularly <i>via</i> D835 activation-loop variants and the F691L gatekeeper substitution, limits durability. Among the major therapeutic classes, type I inhibitors bind the active (DFG-in) conformation, whereas type II inhibitors stabilize the inactive (DFG-out) state. In contrast, irreversible covalent inhibitors target reactive cysteine residues within the kinase domain. Collectively, these approaches represent complementary therapeutic strategies with distinct resistance liabilities and structural design considerations. This review integrates structural biology with medicinal-chemistry evidence across type I, type II, irreversible, and dual-modality FLT3 inhibitors, analyzing how hinge contacts, back-pocket occupancy, and warhead placement govern activity across wild-type and mutant FLT3. We map resistance-defining residues (<i>e.g.</i> F691, D835, N676, N701) and design tactics to preserve potency against resistant variants. We also summarize combination therapy that augments selective FLT3 blockade and outline PROTAC approaches that induce FLT3 degradation, positioning these modalities as alternatives when single-molecule polypharmacology is constrained. Finally, we catalogue dual-target FLT3 chemotypes, highlighting examples that retain activity against F691L and D835 in cellular systems and xenografts. Overall, this review provides a section-by-section guide covering FLT3 structure and mutation hotspots, analyses of type I, type II, and irreversible inhibitors, dual-modality designs, combinations, PROTACs, and future perspectives. It links binding mode, covalent engagement, and second-target selection to recurrent resistance biology to guide more resilient FLT3-targeted therapies for high-risk AML.

Pancreatic cancer responds poorly to conventional chemotherapy, largely because of the pronounced resistance of tumor cells to chemotherapy-induced apoptosis. Ferroptosis, a non-apoptotic form of programmed cell death, has emerged as a promising strategy to overcome this resistance. However, its therapeutic efficacy is often limited by insufficient hydrogen peroxide (H₂O₂) and excessive glutathione (GSH) in the tumor microenvironment (TME). Herein, we developed a nanoplatform, HM-MnO₂@DOX/CaO₂@PDA/HA (HMDCPH), using hollow mesoporous manganese dioxide (HM-MnO₂) as a carrier to co-deliver doxorubicin (DOX) and calcium peroxide (CaO₂). The crosslinked PDA/HA shell enhanced both the tumor-targeting capability and biocompatibility of the nanoplatform. In the TME, HM-MnO₂ depleted GSH and promoted reactive oxygen species (ROS) generation, whereas CaO₂ decomposition generated H₂O₂ and released Ca²⁺, inducing mitochondrial calcium overload and further aggravating oxidative stress. These synergistic effects enhanced lipid peroxidation (LPO) and exacerbated ferroptosis-related oxidative damage. Moreover, the near-infrared (NIR)-triggered photothermal effect further strengthened the antitumor efficacy of HMDCPH. In addition, nanoplatform degradation released Mn²⁺, enabling T₁-weighted magnetic resonance imaging (MRI). Collectively, this study presents a synergistic nanotherapeutic strategy that integrates chemotherapy, photothermal therapy, and ferroptosis-related mechanisms to overcome chemoresistance in pancreatic cancer.

<b>Objectives:</b> Transcriptomic profiling has enabled the classification of Diffuse Large B-Cell Lymphoma (DLBCL) into distinct subtypes, such as Germinal Center B-cell-like (GCB) and Activated B-cell-like (ABC), primarily in HIV-negative patients. However, HIV-associated DLBCL may follow different molecular mechanisms due to immune dysregulation. This study aimed to characterize the transcriptomic landscape of HIV-related DLBCL to identify distinct subtypes and deregulated pathways with potential theranostic implications. <b>Methods:</b> Twelve formalin-fixed, paraffin-embedded DLBCL samples from HIV-positive patients were analyzed using Agilent's microarray. Quantile normalization and unsupervised hierarchical clustering were performed to classify tumors based on gene expression profiles. <b>Results:</b> Two distinct transcriptomic subgroups were identified. <i>TP53</i> and <i>BCL7A</i> were overexpressed in cluster I, while <i>BCL2</i> was overexpressed in cluster II. Notably, the "immune system development" pathway was under expressed in cluster I compared to cluster II. <b>Conclusions:</b> Our findings reveal two molecularly distinct subtypes of HIV-associated DLBCL, likely driven by differences in tumor microenvironment and immune status. These transcriptomic profiles may guide future targeted therapies. Further validation in larger cohorts and integration with proteomic and clinical data are warranted to develop a comprehensive theranostic framework.

<b>Background:</b> Depression-related liability is frequently accompanied by reduced physical function, yet the shared genetic architecture linking mood-related traits and physical-function decline remains incompletely characterized. <b>Methods:</b> We applied genomic structural equation modeling to European-ancestry GWAS summary statistics for five constituent phenotypes: depressive symptoms, depression diagnosis, grip strength, appendicular lean mass, and walking pace. A Depression-Physical Function shared genetic factor was constructed as a cross-trait genetic covariance dimension and evaluated using LDSC-based validation and leave-one-trait-out sensitivity analyses. We then performed factor GWAS, FUMA locus annotation, Bayesian fine-mapping, MAGMA gene-based analysis, transcriptome-wide association analysis, pathway enrichment, CELLECT/MAGMA cell-type specificity analysis, partitioned heritability analysis, and gsMap spatial transcriptomic mapping. <b>Results:</b> The shared factor showed good model fit and retained 755,397 quality-controlled variants for downstream analysis. The factor was positively genetically correlated with depression-related traits and negatively correlated with physical-function-related traits. FUMA identified 245 genome-wide significant SNPs, 44 lead SNPs, and 38 genomic risk loci, with 127 positional mapped genes. Fine-mapping prioritized one high-confidence locus. MAGMA identified 19 Bonferroni-significant genes and 326 FDR-significant genes, while TWAS identified 322 FDR-significant expression-associated genes. Integrating FUMA positional mapping, MAGMA gene-level association and TWAS expression-level association prioritized eight convergent genes: TMEM106B, CENPW, DRD2, LRFN5, NCAPG, DCAF16, SGIP1, and FAM120A. Functional enrichment highlighted postsynaptic structure, neuron spine, synaptic plasticity, and synapse organization. CELLECT/MAGMA prioritized brain non-myeloid neurons and glial populations, with additional endocrine-metabolic and immune-hematopoietic signals. Spatial transcriptomic mapping localized top signals to brain and spinal cord regions in the embryonic neuro-muscle reference. Partitioned heritability analysis showed enrichment in conserved, intronic, promoter, and chromatin-related genomic annotations. <b>Conclusions:</b> These findings support a shared polygenic covariance dimension linking depression-related liability with reduced physical-function-related genetic propensity. Downstream analyses prioritized candidate loci, genes, and biological contexts, with enrichment patterns consistent with neuronal, synaptic, and regulatory genomic processes.

Early childhood exposure to bisphenol A (BPA) is closely associated with autism spectrum disorder (ASD), though the precise molecular mechanisms remain unclear. To investigate this, we employed an integrated computational approach combining network toxicology, molecular docking, and molecular dynamics simulation. Potential targets of BPA and ASD-related genes were collected from multiple databases, identifying 57 overlapping targets. Protein-protein interaction network analysis highlighted 16 core targets among them. Gene Ontology and KEGG pathway enrichment analyses indicated these targets are primarily involved in synaptic transmission, GABAergic signaling, and neuroactive ligand-receptor interactions. Molecular docking demonstrated potential binding between BPA and several core targets, including estrogen receptor 1 (ESR1), gamma-aminobutyric acid type A receptor subunit beta-2 (GABRB2), and amyloid precursor protein (APP), with binding energies below -5 kcal/mol. The stability of the BPA-ESR1 complex was further supported through 100 ns molecular dynamics simulation. These results suggest that BPA may contribute to ASD by disrupting neuroendocrine pathways and synaptic function via interactions with key targets such as ESR1.

Human papillomavirus (HPV)-associated cervical lesions remain a significant disease burden and minimally invasive blood-based biomarkers could complement cytology and HPV testing. This study aimed to characterize the proteomic composition of plasma-derived CD9⁺ small extracellular vesicles (sEVs) across the morphological spectrum of HPV-associated cervical disease, from histologically normal (NILM) through low-grade (LSIL) and high-grade (HSIL) lesions to invasive squamous cell carcinoma (SCC). Plasma samples from 34 women (NILM, LSIL, HSIL, SCC) were pooled per group, and CD9⁺ sEVs were isolated using an electrochemically controlled immunoaffinity capture method, followed by nanoparticle tracking analysis, transmission electron microscopy, Western blotting, and label-free LC-MS/MS proteomic profiling. The core sEV proteome comprised 258 shared proteins. LSIL showed the most pronounced changes with broad enrichment of complement and coagulation components and acute-phase reactants alongside depletion of immunoglobulin chains and complement C1r-like protein (C1RL). HSIL exhibited few differential proteins, dominated by neutrophil degranulation and retinoid metabolism pathways. SCC demonstrated extensive cargo depletion (22 downregulated proteins) and a nearly sevenfold upregulation of C1RL. Five proteins (including immunoglobulin chains and GPLD1) correlated positively with lesion severity. Pathway analysis consistently implicated platelet activation, lipoprotein remodeling, and insulin-like growth factor signaling. We conclude that plasma CD9⁺ sEVs carry stage-specific proteomic signatures distinguishing HPV-associated cervical lesions, with C1RL emerging as a candidate biphasic marker warranting further validation.