搜索结果：Frontiers in cell and developmental biology

This study aims to elucidate the role of Enterococcusin the progression from inflammatory bowel disease to colorectal cancer (CRC), with a focus on identifying key metabolites and host genes regulated by Enterococcusand their influence on CRC development. Using the database gutMGene, gutMDisorder and MACdb, we mined the key metabolites and human genes. We acquired the activated genes (panel 1) and inhibited genes (panel 2), and metabolite associated genes (MAGs, panel 3). Subsequent analyses included protein-protein interaction (PPI) network construction, functional enrichment, differential expression and survival analysis in CRC, and immune infiltration assessment. In vitro experiments validated the regulatory effects of E. faecalisand its key metabolites on candidate genes. Single-cell RNA sequencing (scRNA-seq) was used to dissect cell-type-specific expression patterns within the tumor microenvironment. We screened 12 activated genes (Panel1: IL11, IL24, IFNG, IL10, IL12B, IL1B, IL6, TNF, ANGPTL4, CXCL10, PLIN2, and PPARG) and four inhibited genes (Panel 2: CXCL8, IL6, TNF, and PDCD6IP). Three metabolites were found important in CRC development: agmatine, formate, and levodopa, linking with 28 MAGs. In particular, IL10, IL11, CXCL10, IL1B, and IFNG are protective genes in CRC; and there are four MAGs associated with CRC PFS, and they are all survival-risk genes: COMT, PRL, EDNRA, and MAPK3. Experimental validation showed that E. faecalis significantly upregulated the level of IL-10 and IL-1B in CRC, while its metabolites agmatine and levodopa markedly induced the expression of the survival-risk gene MAPK3. scRNA-seq revealed cell-type-specific expression patterns, where IL1B was significantly upregulated in both tumor epithelial and myeloid cells, and IL10 was specifically elevated in tumor epithelial cells. In contrast, MAPK3 exhibited divergent trends, showing downregulation in tumor epithelial cells but significant upregulation in myeloid cells. Enterococcus exhibits a dual role in colitis-associated CRC, correlating with both tumor-suppressing and tumor-promoting effects. It may activate protective immune genes while its metabolites, agmatine and levodopa, alter the expression of oncogenic MAGs. The findings highlight the complex metabolite-host gene networks driven by Enterococcusand suggest estradiol and sodium arsenite as potential adjuvant therapies, offering new insights into precision intervention for CRC.

Accumulating evidence suggests reciprocal risk factors between periodontitis (PD) and systemic sclerosis (SSc). Ferroptosis, an iron-dependent and immune-related form of cell death, has been implicated in both diseases, yet its shared molecular mechanisms remain largely unclear. Bidirectional Mendelian randomization (MR) analysis was first conducted to evaluate the potential causal relationship between PD and SSc. Gene expression datasets for PD and SSc were retrieved from the GEO database, and ferroptosis-related genes were obtained from FerrDb. Differential expression and WGCNA identified common ferroptosis-related differentially expressed genes (Co-FRDEGs) and common ferroptosis-related module genes (Co-FRMGs). Functional enrichment analyses were subsequently performed. The intersection of Co-FRDEGs and Co-FRMGs yielded candidate genes, which were further screened by three machine learning algorithms (LASSO, SVM-RFE, and Random Forest) to identify shared hub genes. Immune infiltration, single-cell RNA sequencing, regulatory network analysis (TF-miRNA), and drug prediction with molecular docking were further performed. In addition, preliminary in vitro experiments were conducted to validate the expression and potential ferroptosis-associated roles of the identified hub genes. MR revealed an asymmetric causal association between PD and SSc. A total of 28 Co-FRDEGs and 63 Co-FRMGs were identified, and their intersection yielded nine candidate genes. Machine learning analysis predicted FNDC3B and NNMT as shared hub genes, exhibiting good diagnostic performance (AUC >0.75) in both discovery and validation cohorts. Immune infiltration analysis revealed multifaceted immune dysregulation in both diseases, while single-cell analysis confirmed cell type-specific expression of the two hub genes. Regulatory network analysis predicted GTF2E2 and three miRNAs as potential co-regulators. Drug prediction and molecular docking suggested thapsigargin as a potential lead compound. Furthermore, in vitro experiments demonstrated that FNDC3B and NNMT were significantly upregulated in PD- and SSc-like cellular models, and their silencing alleviated ferroptosis-associated cellular injury. This study highlights potential shared ferroptosis-related genes, regulatory networks, and candidate therapeutic compounds associated with PD and SSc, providing new insights into their molecular connections. However, as these findings are largely derived from bioinformatics analyses with preliminary experimental validation, further mechanistic and clinical studies are required to confirm them.

Aging is intimately associated with multisystem functional decline and an increased risk of chronic diseases. A pivotal cytological basis underlying this process is the progressive dysregulation of the mitochondrial quality control (MQC) network. Emerging evidence suggests that MQC is not a singular process but rather a multitiered synergistic system encompassing mitochondrial biogenesis, dynamic remodeling, selective autophagy (mitophagy), proteostasis maintenance, and coordinated mitochondrial-organelle communication. This integrated network is critical for preserving cellular energy homeostasis, redox balance, and stress tolerance. During aging, impairments in mitochondrial genomic coordination, network topology, autophagic flux, and protein import and folding collectively contribute to bioenergetic decline, chronic low-grade inflammation, and metabolic imbalance. As a safe and sustainable nonpharmacological intervention, regular exercise systematically remodels MQC structure and function by integrating signaling axes such as AMPK, SIRT1, and p38 MAPK, thereby promoting coordinated mitochondrial renewal and partially reversing aging-associated mitochondrial dysfunction. On the basis of a systematic elucidation of the core mechanisms of MQC and its dysregulation during aging, this review highlights the differential regulatory effects of distinct exercise modalities-specifically endurance training, high-intensity interval training (HIIT), and resistance training-on mitochondrial dynamics, autophagic flux, proteostasis, and mitochondrial turnover. Furthermore, the intrinsic associations among exercise-MQC coupling, inflammatory responses, metabolic imbalances, and emerging peripheral biomarkers are explored. Finally, current research limitations and challenges in clinical translation are analyzed, and future research directions regarding dose-response relationships, multimodal exercise prescriptions, personalized strategies, and systemic integrated regulation are proposed. This review aims to provide a refined theoretical basis for optimizing exercise-based anti-aging interventions.

Adult hippocampal neurogenesis (AHN) supports learning, memory, and emotional regulation, and is regulated by intrinsic and extrinsic factors. Dopamine influences neurogenesis in animal models, but its direct effects on human hippocampal progenitors and receptor-specific mechanisms remain unclear. This study examined the dose-dependent effects of dopamine on proliferation, differentiation, and survival of human hippocampal progenitor cells (HPC0A07/03) in vitro, and assessed dopamine D4 receptor (DRD4) involvement. Cells were treated with dopamine (1-150 µM) under proliferation and differentiation conditions, with DRD4 modulated via selective agonist, antagonist, or combined treatment. Proliferation (Ki67), stemness (SOX2, Nestin), neuronal differentiation (DCX, MAP2), apoptosis (CC3), total cell counts, and morphology (cytoplasmic area) were assessed using immunocytochemistry, alongside targeted gene expression analysis of cellular stress- and neurogenesis-related pathways. Treatment with supraphysiological dopamine concentration (150 µM) significantly reduced cell counts during differentiation and decreased SOX2 expression during proliferation, suggesting impaired survival and reduced stemness. Complementary transcriptional changes supported a stress-associated cellular response at high dopamine concentrations. Elevated dopamine (150 µM) also increased cytoplasmic area in immature DCX+ neurons during the differentiation phase, suggesting altered morphological maturation. Moderate dopamine concentration (30 µM) showed a trend toward increased proliferation and higher cell counts. No significant changes occurred for other markers or following DRD4 modulation. These findings indicate that dopamine's effects on human hippocampal progenitors are dose-dependent: supraphysiological levels may compromise survival and progenitor identity, potentially via stress-related mechanisms, whereas moderate levels may support neurogenic processes. Understanding this dose-dependent balance has implications for neurological and psychiatric disorders involving dopaminergic dysregulation.

Cancer stem cells (CSCs) play a central role in tumor initiation, progression, recurrence, and therapy resistance. Their abilities for self-renewal, multi-lineage differentiation, and strong resistance make conventional chemotherapy, targeted therapy, and radiotherapy insufficient to completely eradicate tumors. In recent years, circular RNAs (circRNAs), a class of novel non-coding RNAs, have been shown to regulate CSC properties through multiple mechanisms, including acting as miRNA sponges, interacting with proteins, modulating signaling pathways, and encoding small peptides. Accumulating evidence indicates that circRNAs are aberrantly expressed in CSCs across various tumor types, including liver cancer, lung cancer, breast cancer, gastric cancer, prostate cancer, ovarian cancer, glioma, and acute myeloid leukemia, influencing stemness and drug sensitivity via specific signaling pathways or regulatory networks. CircRNAs have potential as biomarkers for diagnosis, prognosis, and therapy resistance prediction, as well as promising therapeutic targets. Strategies targeting oncogenic circRNAs, such as siRNA or shRNA delivered via liposomes, can effectively suppress CSC stemness and resistance and may be combined with chemotherapy, targeted therapy, or immunotherapy. Despite challenges such as incomplete mechanistic understanding, CSC heterogeneity, and limited clinical validation, advances in single-cell sequencing, circRNA interference, and nanocarrier delivery provide new opportunities for clinical translation. Overall, circRNAs play critical roles in maintaining CSC stemness, modulating drug resistance, and promoting tumor progression, offering novel avenues for overcoming therapy-resistant CSCs and for early diagnosis, prognosis assessment, and personalized treatment.

Congenital cataract (CC) is a time-critical cause of preventable childhood visual impairment. After diagnosis, parents frequently experience uncertainty and increasingly seek guidance online. The safety, readability, and counseling quality of large language models (LLMs) responses for CC remain insufficiently benchmarked, particularly for explanations involving lens development, etiology, and genetic risk. We performed a cross-sectional comparative evaluation of five publicly accessible Chinese conversational LLMs (ChatGPT-5.2, Gemini 3 Pro, DeepSeek-V3.1, Doubao, and Kimi K2). Thirty standardized parent-facing CC questions were developed by senior ophthalmologists and mapped to five domains, with specific incorporation of scenarios requiring translation of lens developmental pathology and genetic counseling knowledge. Two researchers independently performed standardized zero-shot querying and response recording under identical conditions. Output efficiency and textual structure were extracted. Two blinded ophthalmologists rated each response on a 5-point Likert scale across Accuracy, Logic, Coherence, Safety, and Content Accessibility; inter-rater agreement was assessed using quadratic weighted Cohen's kappa. Group differences were tested using ANOVA or Kruskal-Wallis H tests with Bonferroni-corrected pairwise comparisons. Significant between-model differences were observed in output efficiency and text characteristics (all P < 0.001). ChatGPT-5.2 was fastest (17.94 &#xb1; 5.11), whereas DeepSeek-V3.1 and Kimi K2 were slowest (41.46 &#xb1; 3.22 and 40.02 &#xb1; 4.67). DeepSeek-V3.1 generated the longest responses (1,456.93 &#xb1; 224.99 words) and Kimi K2 the shortest (640.83 &#xb1; 252.95). ChatGPT-5.2 showed the strongest tendency toward structured/tabular output [2.00 (1.00, 2.00)] followed by Gemini 3 Pro [1.00 (1.00, 1.25)], while the other models rarely produced tables. Quadratic weighted Cohen's kappa indicated good inter-rater reliability (0.686-0.767). Content quality differed significantly across models (Accuracy H = 41.15, Logic H = 32.95, Content accessibility H = 41.33; all P < 0.001). ChatGPT-5.2 and Gemini 3 Pro achieved higher overall profiles and did not differ significantly from each other, whereas Kimi K2 scored lower on multiple dimensions. LLM performance in translating lens developmental pathology and genetics for CC parent counseling is model-dependent. Longer outputs did not necessarily translate into higher quality; structured presentation was more closely associated with better safety and accessibility. These findings provide quantitative benchmarks for safer, parent-centered deployment of LLMs in pediatric ophthalmology education and support more reliable translation of complex disease-related knowledge into actionable parent guidance.

Digestive tract tumors (DTT), particularly gastric cancer (GC) and colorectal cancer (CRC), remain among the leading causes of cancer-related morbidity and mortality worldwide. Accumulating epidemiological evidence indicates that patients with chronic kidney disease (CKD) exhibit a significantly increased risk of developing gastrointestinal malignancies and experience worse clinical outcomes. However, the biological mechanisms underlying this association have not been comprehensively synthesized. In this review, we integrate clinical and experimental evidence to delineate how CKD functions as a systemic pro-tumorigenic condition rather than a passive comorbidity. We highlight three interrelated mechanistic axes linking CKD to DTT: (i) persistent systemic inflammation and oxidative stress, (ii) metabolic and endocrine dysregulation driven by uremic toxin accumulation, vitamin D deficiency, and mineral imbalance, and (iii) immune perturbations associated with dialysis modalities and post-transplant immunosuppression. These processes converge to disrupt gastrointestinal barrier integrity, reshape the gut microbiota, impair antitumor immune surveillance, and promote malignant transformation and tumor progression. Importantly, we discuss how CKD-specific interventions, including dialysis strategies, kidney transplantation, dietary management, and modulation of gut microbiota, may further modify gastrointestinal cancer risk. Finally, we propose CKD-oriented preventive and screening strategies for GC and CRC, emphasizing the need for risk stratification based on renal function, proteinuria, and metabolic profiles. By framing CKD as an active driver of gastrointestinal carcinogenesis, this review provides a novel integrative framework that synthesizes interconnected mechanistic pathways and explicitly links them to CKD-specific clinical management strategies, a translational perspective that informs early detection, prevention, and integrated care of DTT in patients with CKD.

Pre-mRNA splicing is a fundamental step in eukaryotic gene expression, carried out by the spliceosome. This large and dynamic ribonucleoprotein complex undergoes extensive structural rearrangements during each splicing event. Similarly, ribosome biogenesis is a highly regulated process that requires precise control at every stage, from the transcription of pre-rRNA through its chemical modification and cleavage to the final assembly of mature ribosomal subunits. Central to the regulation of both pre-mRNA splicing and ribosome biogenesis are RNA helicases and their cofactors, notably G-patch proteins. The predominance of G-patch proteins in eukaryotes underscores their evolutionary importance in the increasing complexity of RNA processing and ribosome biogenesis. This review summarizes recent findings on the molecular functions and regulatory roles of various G-patch proteins in the yeasts S. cerevisiae and S. pombe, as well as in humans. Growing evidence indicates that these proteins act as critical cofactors of RNA helicases involved in splicing, facilitating the dynamic transitions required for spliceosome activation, catalysis, and disassembly. Beyond splicing, these proteins also contribute to the regulation of ribosome biogenesis and other aspects of RNA metabolism. Dysregulation or mutation of G-patch proteins have been shown to cause aberrant mRNA maturation, altered splicing patterns, impaired ribosome assembly, and genomic instability. Such perturbations are associated with a range of human diseases, including cancer progression. Despite the essential roles of G-patch proteins in regulating pre-mRNA splicing and ribosome biogenesis, the precise molecular functions and interaction networks of many G-patch proteins remain poorly understood. Future studies aimed at elucidating the mechanisms by which these proteins coordinate RNA processing and ribosome biogenesis are therefore essential. Such investigations may help uncover the molecular basis of G-patch protein-associated diseases and reveal new potential targets for therapeutic intervention.

Erectile dysfunction (ED) is increasingly prevalent worldwide, arising from complex interactions between genetic susceptibility and environmental exposure. Real-world exposure involves complex chemical mixtures that may induce synergistic toxicity, which traditional methods struggle to elucidate. This study integrates multi-omics data with reverse network toxicology to systematically identify causal molecular targets and environmental pollutants underlying ED risk, thereby clarifying their mechanisms. We performed summary data-based Mendelian randomization (SMR) integrating proteomic (pQTL), transcriptomic (eQTL), and DNA methylation (mQTL) data to identify plasma proteins, gene expression levels, and methylation sites causally linked to ED, with false positives excluded via HEIDI tests. The identified targets were used to screen environmental pollutants in the Comparative Toxicogenomics Database Toxicity was predicted using ADMETlab 3.0 and ProTox-III, followed by molecular docking to validate interactions. Functional assays in HUVECs assessed the role of FIS1 and the effects of benzo[a]pyrene. pQTL-SMR analysis identified 28 plasma proteins significantly associated with ED risk, with consistent effects in both discovery and validation cohorts. Integrated eQTL and mQTL analyses further prioritized FIS1, TNFSF12, and CNP as core targets linked to ED at the protein, gene expression, and methylation. Multi-omics evidence revealed that distinct methylation sites within these genes differentially regulate transcription and translation, exerting different impacts on ED. Using these targets, we screened four environmental pollutants-bisphenol F, tetrabromobisphenol A, benzo[a]pyrene, and chlorpyrifos-as potential regulators. Toxicity predictions indicated mutagenic, cytotoxic, or endocrine-disrupting potential for these compounds. Molecular docking confirmed stable binding to the target proteins (binding free energy &#x394;G < -5.0&#xa0;kcal/mol). In vitro experiments showed that inhibition of FIS1 expression suppressed HUVEC proliferation and mitochondrial function, and exposure to benzo[a]pyrene similarly impaired these processes and reduced FIS1 expression. This study delineates a potential "environmental pollutant-molecular target-ED" mechanistic pathway, offering new insights into the environmental etiology of ED and establishing a theoretical basis for risk assessment and targeted prevention strategies.

Dysregulated dsRNA editing and RNA metabolism in cancer contribute to immune evasion, highlighting the critical role of RNA structural regulation in disease. Intracellular RNA structures regulate gene expression, innate immunity, genome stability, and cell fate. Among these, double-stranded RNA (dsRNA) is particularly important; exogenous dsRNA typically originates from viral infection, whereas endogenous dsRNA arises from repetitive elements or transcriptional errors, allowing cells to distinguish "self" from "non-self." The RNA-editing enzyme Adenosine Deaminase Acting on RNA 1 (ADAR1) prevents inappropriate innate immune activation by catalyzing adenosine-to-inosine (A-to-I) editing of endogenous dsRNA. RNA helicases complement this function by remodeling RNA structures and resolving nucleic acid hybrids, maintaining RNA homeostasis and immune surveillance. Recent studies have revealed an interplay between ADAR1 and RNA helicases that regulate dsRNA immunogenicity and R-loop dynamics, establishing this network as a key determinant of tumor immunity. Dysregulated RNA editing and structural regulation in cancer further underscore the potential of targeting these pathways therapeutically, providing strategies beyond conventional gene- or protein-centered approaches. In this review, we summarize current insights into how ADAR1 and RNA helicases control RNA structure, emphasize their roles in innate immune sensing, and discuss emerging approaches to modulate RNA editing and RNA architecture for therapeutic benefit. Taken together, research in this area positions RNA structural control as a central determinant of immune homeostasis and a promising frontier in cancer therapy.

Brain metastasis (BM) remains a severe and fatal complication in patients with lung cancer (LC), presenting a major therapeutic challenge. Although epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) have emerged as a cornerstone of targeted therapy, their clinical efficacy is often limited by the inevitable development of drug resistance. We initially constructed a general atlas of the tumor microenvironment (TME) in LCBM lesions by integrating single-cell RNA sequencing (scRNA-seq) data. The sensitivity of each cell cluster to EGFR-TKIs was assessed by the "Beyondcell" method. By performing high-dimensional Weighted Gene Co-expression Network Analysis (hdWGCNA), we identified hub genes within an EGFR-TKI resistance-associated cell cluster. Finally, the functional role of the most promising candidate, ACTN1, was further investigated in a constructed osimertinib-resistant LC cell line. We identified a malignant and therapy-resistant ACTN1+ epithelial cell subcluster. Both signaling and functional enrichment analyses demonstrated marked activation of PI3K-Akt and IL-17 signaling pathways in ACTN1-high patient subgroups. Finally, we applied machine learning methods to the ACTN1-related genes to select prognostic factors. In vitro experiments confirmed the pro-resistance and pro-metastatic functions of ACTN1 in osimertinib-resistant LC cells. ACTN1 was discovered to induce malignant progression and formation of EGFR-TKI resistance. Targeting ACTN1-related pathways may provide novel insights to treat LCBM and overcome intracranial EGFR-TKI resistance.

Lipedema is a chronic, female-predominant disorder of subcutaneous adipose tissue characterized by disproportionate fat expansion, pain, and fibrosis. Despite its high prevalence, the cellular mechanisms underlying lipedema remain poorly understood. While the clinical features have been extensively described, its biology of adipose tissue dysfunction and aberrant intercellular communication is still unclear. In comparison to obesity, lipedema is marked by local dysregulation of adipocyte-stromal and adipocyte-vascular interactions. In this hypothesis perspective, we discuss emerging mechanistic concepts from a cell biology perspective that are particularly relevant to lipedema, focusing on (i) organelle contact site dynamics in adipocytes and their role in lipid handling and stress adaptation; (ii) extracellular vesicle (EV)-mediated crosstalk between endothelial cells, adipocytes, and immune cells as a driver of localized inflammation and fibrosis; and (iii) estrogen-linked signaling pathways that may imprint EV cargo and cellular behavior in a sex-specific manner. By integrating these perspectives, we highlight open experimental settings and mechanistic parallels to other adipose tissue pathologies that help understanding lipedema as a distinct cellular and molecular entity. Investigating how organelle biology, extracellular vesicles communication and hormonal context intersect in adipose tissue may uncover novel biomarkers and therapeutic entry points for this long-neglected condition.

Peripheral nerve injuries remain a major clinical challenge, often leading to long-term motor and sensory deficits. Stem cell-based and cell-free approaches, combined with biomaterial scaffolds, have emerged as promising strategies for nerve repair. This study evaluated the regenerative potential of human dental pulp stem cells (hDPSCs), their conditioned medium (hDPSCs-CM), and the combination of hDPSCs with olfactory mucosa mesenchymal stem cell conditioned medium (OM-MSCs-CM) in a rat model of sciatic nerve neurotmesis repaired with a chitosan-based nerve guide conduit (Reaxon&#xae;). Twenty-seven rats were allocated into experimental groups, including an uninjured control (contralateral limb), an end-to-end neurorrhaphy surgical control group, and treatment groups repaired with Reaxon&#xae; alone or combined with hDPSCs, hDPSCs-CM, or hDPSCs with OM-MSCs-CM suspended in Matrigel&#xae;. Following nerve transection, a 9-10 mm sciatic nerve gap was created. Functional recovery was monitored over 20 weeks through motor, nociceptive, behavioral, gait, stereological, histomorphometric, and electrophysiological evaluations. All treatments promoted progressive motor recovery and partial restoration of nociceptive function compared to the untreated condition, although the magnitude of improvement differed among groups. The hDPSCs-CM-treated group (CMDP) showed the most favorable overall outcomes, including the lowest muscle mass loss, higher compound muscle action potential amplitudes, and functional indices approaching control values, indicating enhanced reinnervation and neuromuscular preservation. Histomorphometric and stereological analyses confirmed active regeneration across all groups, characterized by microfasciculation and thinner myelin sheaths typical of regenerating fibers. Despite incomplete recovery, the combination of biological therapies with chitosan conduits provided an effective environment for axonal regrowth and functional improvement. These findings highlight the relevance of CMDP as a potent biological adjunct in peripheral nerve repair and support the development of cell-free, clinically translatable strategies for neuroregeneration.

Chemotherapy remains a cornerstone treatment for various malignancies; however, its efficacy is often limited by the development of drug resistance, which increases the risk of tumor recurrence and metastasis. Chemoresistance arises from multiple mechanisms, including enhanced drug efflux, apoptosis inhibition, increased DNA damage repair, and the maintenance of cancer stem cells (CSCs). Recent studies have revealed that epigenetic alterations play a critical role in chemoresistance. DNA methylation, histone modifications, and non-coding RNAs contribute to resistance by regulating gene expression, signaling pathways, and CSC properties. RNA epigenetic modifications, such as N6-methyladenosine (m6A), N4-acetylcytidine (ac4C), and 5-methylcytidine (m5C), regulate mRNA stability, splicing, and translation efficiency, thereby sustaining CSC self-renewal and promoting resistance. Epigenetic-targeted agents, including DNA methyltransferase inhibitors (DNMTi), histone deacetylase inhibitors (HDACi), and emerging inhibitors targeting RNA-modifying enzymes, have demonstrated the potential to reverse resistance, suppress CSC traits, and enhance chemosensitivity in vitro and in vivo. Combining epigenetic drugs with conventional chemotherapy enables multi-level intervention in resistance mechanisms, significantly improving therapeutic outcomes and offering new avenues for personalized cancer treatment. Future studies should focus on developing precise biomarkers, optimizing combination strategies, and conducting clinical validation to advance the application of epigenetic interventions in chemoresistant cancers.

Head and neck squamous cell carcinoma (HNSCC) exhibits heterogeneous responses to immunotherapy, primarily determined by the tumor immune microenvironment (TME). Accordingly, there is an urgent need for reliable biomarkers that can effectively distinguish between immune "hot" and "cold" tumors and predict prognosis and immunotherapy response. We integrated transcriptomic and clinical data from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) cohorts to identify immune-related genes (IRGs) associated with TME phenotypes. Unsupervised clustering was used to define immune-hot and immune-cold subtypes. A six-gene prognostic signature (PYGL, SFRP1, FGD3, OLR1, DUSP9, and MASP1) was developed using Cox and Least Absolute Shrinkage and Selection Operator (LASSO) regression, which was validated across four cohorts and evaluated for its association with immune infiltration, tumor mutation burden (TMB), and immunotherapy response. The functional role of PYGL was assessed via siRNA knockdown in HNSCC cell lines. The signature demonstrated good performance in stratifying patients into high- and low-risk groups, characterized by distinct survival, immune landscapes, and genomic mutation profiles. High-risk patients showed an immunosuppressive TME, higher TMB, and poorer response to immune checkpoint blockade. PYGL knockdown suppressed proliferation, migration, and clonogenicity. Transcriptomic analysis revealed downregulation of MYC/E2F targets and EMT pathways, alongside concurrent activation of interferon and immune response programs. This IRG-based signature offers a clinically translatable tool for prognostication and immunotherapy selection in HNSCC. PYGL represents a key oncogenic driver and potential therapeutic target linking cellular metabolism to tumor progression and immune evasion.

ZFHX3, a multifunctional transcription factor, plays pivotal roles in a variety of physiological and pathological processes, including neuronal differentiation, development, atrial fibrillation, and cancer. Notably, homozygous deletion of Zfhx3 is embryonic lethal, whereas its hemizygous deletion results in reduced neonatal body weight and increased preweaning mortality in mice. Also, a deletion mutation in Zfhx3 is significantly associated with reduced litter size, a key reproduction-related trait in goats, suggesting that ZFHX3 is involved in reproductive development. However, its specific function in female reproduction remains unclear. Given that oocyte meiosis is a fundamental process in female reproduction, we investigated the role of Zfhx3 in this process. We performed siRNA microinjection, immunofluorescence staining, chromosome spreading, Western blotting and other experiments to investigate the function of Zfhx3 in mouse oocyte meiosis. We found that Zfhx3 was present in both the nucleus and cytoplasm in GV oocytes and became predominantly localized to the cytoplasm after germinal vesicle breakdown. Knockdown of Zfhx3 caused failure of first polar body extrusion due to sustained activation of the spindle assembly checkpoint (SAC). Zfhx3-deficient oocytes were defective in spindle assembly, microtubule-kinetochore attachment, and chromosome segregation during meiosis I, resulting in aneuploidy in MII oocytes. These defects could be ameliorated by ectopic expression of Zfhx3 mRNA. Our findings provide evidence for an essential role of Zfhx3 in spindle assembly and chromosome segregation during mouse oocyte meiosis I, and provide a mechanistic basis for its mutations in female reproductive disorders.

Metabolic reprogramming is a hallmark of glioblastoma multiforme (GBM), with lactate dehydrogenase A (LDHA) playing a key role in aerobic glycolysis. However, the upstream transcriptional mechanisms driving LDHA overexpression in GBM remain poorly understood. This study aims to investigate the transcriptional regulatory network governing LDHA-mediated glycolysis and to explore the functional roles of KLF1 and NFYC in GBM progression. Bioinformatic analysis of The Cancer Genome Atlas data was performed to assess LDHA, KLF1, and NFYC expression in glioma. glioblastoma multiforme cell lines were used for loss- and gain-of-function studies by siRNA/shRNA knockdown and overexpression, including assessments of glycolytic flux, mitochondrial metabolism, cell proliferation, and apoptosis Metabolic activity was assessed using Seahorse extracellular flux analysis. Transcriptional regulation was evaluated by dual-luciferase reporter and chromatin immunoprecipitation (ChIP) assays. Tumor growth was assessed in a subcutaneous xenograft model. LDHA was significantly upregulated in GBM and associated with poor prognosis. LDHA knockdown suppressed tumor growth, glycolysis, mitochondrial respiration, and induced apoptosis. KLF1 was identified as a direct transcriptional activator of LDHA. NFYC was shown to bind the KLF1 promoter and positively regulate its expression. Functional studies demonstrated that the NFYC-KLF1-LDHA axis promotes GBM cell proliferation, inhibits apoptosis, and enhances glycolytic and mitochondrial metabolism. The oncogenic effects of NFYC were partially reversed by KLF1 knockdown, and vice versa. This study reveals a novel hierarchical transcriptional pathway in which NFYC regulates KLF1, which in turn activates LDHA, driving aerobic glycolysis and tumor progression in GBM. Targeting the NFYC-KLF1-LDHA axis may represent a promising therapeutic strategy for glioblastoma.

The basement membrane is a specialized extracellular matrix that compartmentalizes epithelial and endothelial tissues and provides essential structural and signaling cues for tissue organization. Whereas fibrillar collagens (Col) of the interstitial matrix (such as Col I and Col III) are widely used in tissue modeling, the networking collagens that scaffold the basement membrane, including human Col IV and Col VI, remain difficult to access. Commercial basement membrane surrogates such as Matrigel&#xae; are derived from murine tumors and are ill defined, dilute, variable, and incompatible with animal-free biomanufacturing. Thus, there is a crucial need for human-derived basement membrane matrices that are free of xenogenic contaminants and do not rely on breeding animals. Here, we analyzed whether human mesenchymal stromal cells (MSCs) could serve as a platform to produce self-assembling basement membrane components under chemically defined, xeno-free conditions. MSCs from placental, umbilical cord, bone marrow, and adipose tissues were cultured as three-dimensional spheroids and adherent multilayered sheets. Confocal imaging of whole-mount, decellularized spheroid matrices showed complex networks of fibronectin (FN) and Col IV with topological and organizational features characteristic of basement membrane. Perinatal MSCs produced distinct matrix architectures consisting of apical FN sheets underlaid by continuous Col IV networks. Time-resolved imaging of umbilical cord MSC-derived matrix sheets demonstrated a reproducible sequence of basement membrane assembly that parallels developmental tissue organization. Together, these findings demonstrate that human MSCs cultured entirely without entirely animal-derived components can synthesize functional basement membrane proteins that self-assemble into ordered, tissue-like scaffolds. In this work, we establish MSCs as a scalable, sustainable, and cruelty-free platform for manufacturing human basement membrane matrices for bioengineering and regenerative medicine applications.

Rare diseases, which collectively affecting millions of people worldwide, present unique diagnostic and therapeutic challenges due to their low prevalence and phenotypic heterogeneity. The importance of epigenetic deregulations in the pathophysiology of rare diseases has been highlighted by recent research on neurodevelopmental diseases and congenital malformation syndromes. Among these, abnormalities in histone modifications (especially lysine methylation and acetylation) have emerged as one of the key mechanisms underlying disease phenotypes. Histone-modifying enzyme mutations result in a variety of developmental diseases, including Kabuki, Rubinstein-Taybi and Weaver syndromes, often manifesting as cognitive impairments, craniofacial abnormalities and growth deficiencies. This review explores the functional convergence of genes encoding histone modifiers and their roles in chromatin regulation. It also analyzes the distribution of variants in these genes and their association with overlapping phenotypes across rare diseases. The findings highlight how different variants within the same gene can result in diverse phenotypic outcomes, and how variants in distinct genes may manifest convergent phenotypes underscoring the interconnected nature of epigenetic deregulations and their implications for understanding genotype-phenotype relationships. By focusing on the subunits of key histone-modifying complexes, we also systematically mapped associated Mendelian phenotypes and highlighted a subset of genes not yet linked to defined syndromes but showing strong intolerance to loss-of-function variants, suggesting their potential involvement in undiagnosed or emerging neurodevelopmental disorders.