Search Results (968)

Adult hematopoietic stem cells (HSCs) and bone marrow (BM) mesenchymal stem/stromal cells (MSCs) are essential for lifelong hematopoiesis, skeletal homeostasis, immune competence, and tissue regeneration. The use of electronic cigarettes (E-cigs) among women of reproductive age continues to rise, raising concerns about potential adverse developmental effects; however, the long-term consequences of maternal E-cig vaping on offspring BM stem cell function and hematopoietic homeostasis remain incompletely understood. Here, using a rat model of maternal E-cig exposure (containing nicotine) during gestation, combined with longitudinal in vivo analyses and complementary ex vivo studies of human cells, we show that prenatal E-cig exposure is associated with persistent alterations in offspring BM stem cell function and lineage commitment. Gestational E-cig exposure was associated with expansion of the CD11b/c⁺ myeloid-enriched compartment, increased CD90⁺ stromal cells, and impaired osteogenic differentiation in rat offspring. Complementary experiments using primary human cells showed that nicotine exposure was associated with reduced T-cell proliferation and impaired cytotoxic activity in a proof-of-principle co-culture assay. Mechanistically, transcriptomic profiling followed by Gene Ontology and pathway enrichment analyses identified alterations in molecular programs associated with KLF4–Notch1 signaling, mitochondrial biogenesis, inflammation, and stem cell regulation in the BM of E-cig-exposed rat offspring. Changes in CCL11, FTO, and RUNX2 were additionally associated with an inflammatory and aging-related molecular phenotype that persisted from early life into adulthood, although these findings do not establish a causal CCL11–FTO–RUNX2 signaling axis or direct cellular senescence. Collectively, our study provides a phenotypic and mechanistic framework for understanding how maternal E-cig exposure may influence long-term offspring hematopoietic, skeletal, and immune health while highlighting the need for further studies to establish causal molecular mechanisms and determine their relevance to maternal E-cig use in humans. Full article

Cashmere goats are widely distributed in the cold, arid and semi-arid remote regions of northern China. Their main economic value is the production of precious cashmere fibers. The differentiation of second hair follicle (SHF) stem cells into hair follicle lineages plays a crucial role in SHF regeneration as well as in the morphogenesis and growth of cashmere fibers; however, its precise molecular mechanism is still unclear. In this study, we found that N⁶-methyladenosine (m⁶A)-circHECA recruiting FUS promoted the differentiation of SHF stem cells into hair follicle lineages through stabilizing FOXM1 mRNA in SHF stem cells, thereby activating the NOTCH pathway in cashmere goats. Furthermore, we confirmed that the m⁶A modification of circHECA is required for the FUS/FOXM1-mediated NOTCH signaling pathway to facilitate the differentiation of SHF stem cells into hair follicle lineages via transfecting circHECA m⁶A-deficient mutants. Our results contribute to elucidating the functional mechanism of m⁶A-circHECA in the differentiation of SHF stem cells into hair follicle lineages in cashmere goats. Full article

Background: Homeobox transcription factors regulate developmental programs, cellular plasticity, and tumor progression, yet the role of H2.0-like homeobox (HLX) in prostate cancer (PCa) remains poorly defined. We investigated the clinical significance of HLX and its relationship to the pro-metastatic transcription factor TBX2. Methods: Transcriptomic and clinical datasets from TCGA, MET500, and SU2C/PCF cohorts were analyzed to assess HLX expression, clinicopathologic associations, and its relationship with TBX2. Functional studies in human PCa cell lines included TBX2 gain- and loss-of-function, HLX knockdown, chromatin immunoprecipitation (ChIP), and expression analyses. Shared HLX- and TBX2-associated pathways were evaluated by Reactome enrichment analysis, and Hallmark Gene Set Enrichment Analysis compared castration-resistant prostate cancer (CRPC) bone metastases with high versus low HLX expression (GSE77930; n = 5/group). In vivo relevance was assessed in an orthotopic TBX2 dominant-negative PCa xenograft model. Results: Human PCa datasets showed that HLX expression was elevated in PCa versus normal prostate tissue and associated with higher Gleason grade, lymph node involvement, aggressive molecular subtypes, and shorter disease-free survival. HLX expression also positively correlated with TBX2 across human PCa cohorts. HLX- and TBX2-associated transcriptional programs converged on extracellular matrix organization, cell adhesion, NOTCH, and VEGF-MAPK signaling pathways. Furthermore, HLX-high CRPC bone metastases were enriched for epithelial–mesenchymal transition, NOTCH, TGF-β, inflammatory, angiogenic, hypoxic, and KRAS signaling pathways. Mechanistic studies showed that HLX knockdown suppressed extracellular matrix-associated genes and key NOTCH pathway components. ChIP demonstrated direct TBX2 binding to the HLX promoter, and genetic modulation of TBX2 expression established HLX as a downstream target of TBX2. Consistent with these findings, reduced HLX expression in orthotopic TBX2 dominant-negative xenografts was associated with loss of metastatic progression. Conclusions: HLX is a candidate biomarker of aggressive PCa and a direct transcriptional target of TBX2. These findings identify a previously unrecognized TBX2–HLX regulatory axis associated with metastatic transcriptional programs and aggressive disease in advanced PCa. Full article

Muscle stem cells (MuSCs) are the cellular source for the generation and regeneration of skeletal muscle. Proper muscle growth requires precise control over the differentiation and self-renewal of MuSCs. Signaling systems, such as bone morphogenetic proteins (BMPs) and Notch, suppress the myogenic differentiation of MuSCs. This allows the expansion of the progenitor pool necessary for muscle growth. To better understand the molecular mechanisms and target genes of BMPs during myogenesis, we examined the response of adult mouse MuSCs to BMP6. BMP6 stimulation of freshly isolated MuSCs suppressed myogenic differentiation. Short-term stimulation (one hour) rapidly increased the expression of classical BMP target genes, such as Id1, as well as Notch pathway genes, including Hes1, Hey1, Lfng, and Snai1. We used Cleavage Under Targets and Tagmentation (CUT&Tag) to generate whole-genome binding profiles for pSMAD1/5/9 and SMAD4, which are transcriptional effectors of the BMP pathway. This method detected dynamic binding in promoters and regulatory elements of direct BMP targets, including Notch pathway genes. Our data demonstrate that BMP6 is a potent suppressor of MuSC differentiation and reveal that a subset of well-characterized anti-myogenic genes (i.e., Hes1 and Hey1) are shared targets of the BMP and Notch pathways. Full article

As a high-value meat duck breed, Muscovy ducks exhibit marked individual differences in egg production that limit the economic efficiency of commercial farming. To elucidate the underlying mechanisms, we integrated phenotypic observation, ovarian transcriptome sequencing, and cecal 16S rRNA gene amplicon sequencing to compare high- and low-laying black-feathered Muscovy duck populations. We recorded 40-day egg production, assessed ovarian morphology, quantified gene expression profiles, and characterized microbial community structure. High-laying ducks produced significantly more eggs (24.60 ± 9.22) than low-laying ducks (15.80 ± 6.64, p < 0.01), with larger ovaries and a greater number of follicles at all developmental stages. Transcriptomic analysis revealed 823 DEGs, with KEGG enrichment implicating pathways governing ovarian physiology, including MAPK, Calcium, Notch, and Wnt signaling. Microbial profiling demonstrated significant differences in α/β diversity between groups, with high-laying ducks exhibiting an elevated Firmicutes/Bacteroidota ratio and decreased abundance of Ligilactobacillus. Integrative correlation analysis identified significant associations between Ligilactobacillus abundance and the expression of Arhgap27 and HOXA10, two genes linked to reproductive function, and highlighted the glycolysis/gluconeogenesis pathway as a convergently enriched signaling cascade shared between gut microbiome and ovarian transcriptome. This multi-omics study uncovers the molecular and microbial basis of egg production variation in Muscovy ducks, offering potential targets for improving breeding efficiency through microbiota-directed or gene-based interventions. These findings deepen our understanding of host–microbe interactions in poultry reproduction and identify candidate genes and microbial taxa that may serve as potential targets for future nutritional or genetic interventions to enhance laying performance in commercial duck production. Full article

Atherosclerosis remains the principal underlying cause of myocardial infarction, ischemic stroke and peripheral artery disease, and its progression reflects a complex interplay between lipid accumulation, endothelial dysfunction, chronic vascular inflammation and maladaptive remodeling of the arterial wall. The Notch signaling pathway, an evolutionarily conserved juxtacrine communication system, has emerged as a central regulator of every cell type implicated in atherogenesis, including endothelial cells, vascular smooth muscle cells, monocytes/macrophages and T lymphocytes. Depending on the receptor–ligand pairing, the hemodynamic context and the cellular compartment involved, Notch signaling can be either atheroprotective or atherogenic, a duality that has complicated efforts to translate mechanistic insight into therapy. This review summarizes current knowledge of the molecular architecture of the Notch pathway in the vasculature, dissects its cell type-specific and stage-specific contributions to atherosclerotic plaque initiation, progression, calcification and destabilization, and critically appraises pharmacological strategies designed to modulate Notch activity, including γ-secretase inhibitors, ligand- and receptor-directed monoclonal antibodies, soluble decoy receptors, microRNA-based approaches and drug repurposing strategies such as statins. Particular attention is paid to the cardiovascular toxicities that have emerged from oncology trials of Notch pathway inhibitors, which illustrate both the pharmacological tractability and the narrow therapeutic window of this pathway. We conclude that Notch-directed therapy for atherosclerosis is mechanistically well justified but will require cell type-selective and context-selective delivery strategies to be clinically viable. Full article

Compartment-specific induced pluripotent stem cell (iPSC)-derived cardiomyocytes provide a powerful resource to study cellular and molecular underpinnings of congenital heart disease (CHD) and acquired cardiovascular disease (CVD). Human heart development requires coordinated and complex regulation of key signaling pathways including Notch, BMP, Wnt, Nodal, and Shh during each step of heart morphogenesis. Compartment-specific cardiac cells for modeling the various structures in the heart can be generated by fine tuning these signaling pathways in a sequential manner thereby mimicking spatiotemporal regulation during embryonic heart morphogenesis. In this review, we provide a brief overview of key signaling pathways that are responsible for forming the distinct structures of the heart originating from the first heart field (FHF) and second heart field (SHF). We then summarize recent differentiation protocols that leverage key heart-development related signaling molecules to generate compartment-specific cardiomyocytes and organoids for disease modeling and therapeutic development in cardiovascular disease. Full article

Hidradenitis suppurativa (HS) is increasingly recognized as a disorder of intrinsic dysregulation at the intersection of genetic susceptibility, host–microbial interactions, and hormonal signaling, with select environmental exposures acting as important secondary modifiers. This narrative review synthesizes mechanistic, clinical, and epidemiologic evidence on intrinsic and extrinsic contributors to HS pathogenesis. Genetic susceptibility for HS involves pathways regulating keratinocyte differentiation, epidermal stem cell function, and follicular architecture, including Notch signaling and transcriptional regulators such as SOX9 and KLF5. Microbiome alterations in both lesional and non-lesional skin suggest that early dysbiosis may contribute to follicular occlusion, epithelial disruption, and immune activation. Hormonal signaling, particularly androgen signaling, promotes follicular dysfunction and inflammation during periods of hormonal fluctuation, frequently aligning with the time of disease onset and flares. Environmental exposures vary considerably in the strength of supporting evidence: tobacco use and elevated body mass index have the most robust epidemiologic and mechanistic data, heat and humidity are increasingly recognized as disease activity modifiers, and evidence for air pollution and microplastics is growing. Collectively, these findings support a model in which HS arises from intrinsic follicular dysregulation shaped by genetic, microbial, and hormonal factors, with environmental exposures influencing but unlikely to independently initiate disease. Full article

Vasculogenesis, the de novo formation of blood vessels from endothelial progenitor cells, is increasingly recognized as a critical contributor to tumor vascularization, complementing classical angiogenesis and promoting cancer progression, metastasis, therapeutic resistance, and disease recurrence. This narrative review aims to provide a comprehensive overview of the molecular mechanisms governing vasculogenesis across human malignancies and to discuss their translational relevance for targeted cancer therapy. This is a narrative, mechanistically oriented review rather than a systematic review or meta-analysis. The literature was searched in PubMed, Scopus, and Google Scholar, with the primary search covering publications from January 2010 to February 2026, while seminal earlier studies were included when they established fundamental concepts in vascular biology. Search terms included combinations of “vasculogenesis”, “tumor vasculogenesis”, “angiogenesis”, “endothelial progenitor cells”, “vascular remodeling”, “VEGF”, “HIF-1α”, “Notch”, “Wnt”, “PI3K/AKT”, “TGF-β”, “EndMT”, “mechanotransduction”, “non-coding RNAs”, “tumor microenvironment”, and “cancer”. Evidence was qualitatively appraised and synthesized according to mechanistic relevance and consistency across experimental and clinical studies. The reviewed literature highlights the complex interplay between developmental signaling pathways, endothelial plasticity, mechanical cues, inflammatory mediators, and non-coding RNAs in regulating tumor vasculogenesis. Collectively, these findings suggest that targeting vasculogenesis-related pathways may complement existing anti-angiogenic therapies and represent a promising strategy for improving precision oncology and overcoming treatment resistance. Full article

Colorectal cancer (CRC) remains a major cause of cancer-related mortality, with recurrence and treatment resistance as persistent challenges. Increasing evidence supports the cancer stem cell (CSC) model in CRC, in which a subpopulation of self-renewing colorectal cancer stem cells (CCSCs) sustains tumor growth, metastasis, and therapy resistance. CCSCs are increasingly recognized as key drivers of tumor progression, therapeutic resistance, and relapse, and are increasingly being investigated not only as therapeutic targets but also as biomarkers with diagnostic and prognostic relevance in CRC. Multiple signaling pathways regulate CCSC maintenance and aggressive behavior, including Wnt/β-catenin, Notch, Hedgehog, PI3K/AKT, MAPK/ERK, NF-κB, and TGF-β signaling. These pathways form the basis of therapeutic strategies aimed at modulating stemness-associated pathways, with several CSC-focused agents and pathway inhibitors currently under translational and clinical investigation. In parallel, CCSC surface markers and molecular signatures are being explored as putative tools for early detection, minimal residual disease monitoring, and outcome prediction. However, successful translation of CSC-directed approaches into effective and safe clinical applications remains difficult. Tumor heterogeneity, CSC plasticity, compensatory signaling mechanisms, and the shared regulatory networks between malignant and normal intestinal stem cells pose substantial barriers. In this review, we summarize the molecular mechanisms underlying CCSC biology, discuss their diagnostic and prognostic implications. We also focus on clinical trials that apply CCSC-directed therapeutic, biomarker, and prevention strategies in CRC., Finally, we emphasize the lessons learned and the translational challenges, including the need for rigorously validated, CCSC-specific biomarkers, that continue to shape the development of these therapeutic and biomarker strategies. Full article

Objectives: The aims of this study were to explore the genetic variants identified by Next-Generation Sequencing (NGS) in patients presenting syndromic/non-syndromic cleft palate (CP) associated with tooth agenesis (TA) and to describe the observed phenotype–genotype correlations. Methods: A systematic review exploring PubMed, Scopus and Web of Science was conducted. Data extraction and bias assessment were performed. Results: From 227 screened articles, 8 studies were included. Second premolars were the most frequently missing teeth, followed by central incisors in non-syndromic CP cases. Genetic variants were most commonly reported in IRF6, FGFR1, NOTCH2, CTNND1, ZFHX4 and AXIN2. Several mutations in these genes were associated with syndromic forms such as Pierre Robin Sequence, Kallmann syndrome, and Van der Woude syndrome. Conclusions: This study suggests a potential shared genetic pathway between CP and TA and supports further exploration of TA as a possible clinical indicator of syndromic cases. NGS emerges as a valuable exploratory tool for identifying such associations, though validation in larger patient cohorts remains necessary. Full article

Background: Aberrant DNA methylation is a hallmark of acute myeloid leukemia (AML) and contributes to leukemogenesis, treatment response, and clinical heterogeneity. While genome-wide methylation studies have identified prognostic methylation signatures, the impact of DNA methylation within pharmacologic pathways and AML-relevant disease genes remains incompletely understood. We investigated the association of DNA methylation in genes of pharmacokinetic/pharmacodynamic (PK/PD) pathways of drugs used to treat AML and in myeloid leukemia-related genes with treatment outcomes in pediatric AML. Methods: DNA methylation profiles from 924 pediatric AML patients treated on Children’s Oncology Group trials (AAML1031, AAML0531, and AAML03P1—available publicly) were analyzed as a discovery cohort. A validation cohort included 159 patients treated on the AML02 trial. A total of 2296 variable CpG sites mapping to 65 PK/PD genes and 107 AML biology/leukemia stemness genes were evaluated. Associations between CpG methylation, gene expression, event-free survival (EFS), overall survival (OS), and measurable residual disease after induction I (MRD1) were assessed using Cox proportional hazards, logistic regression, and correlation analyses. Results: Twenty-three CpG sites in PK/PD genes and forty-two CpG sites in AML-related genes were significantly associated with at least one clinical endpoint after Bonferroni correction (p < 2.17 × 10⁻⁵). Hypermethylation of drug transporters ABCA3, ABCC1, and SLC22A1, as well as pharmacologically relevant genes MPO, NOS3, and CTPS1, was associated with inferior survival and/or increased MRD1 positivity. Among AML biology genes, methylation of ETV6, NOTCH1, RUNX1, KIT, MPL, DNMT3A, and DNMT3B demonstrated consistent associations with outcomes across discovery and validation cohorts. Several genes exhibited significant inverse correlations between DNA methylation and gene expression, including MPO, KIT, MPL, SPINK2, and DNMT3B, supporting functional epigenetic regulation. Notably, hypermethylation of ABCA3, MPO, and MPL was reproducibly associated with poor OS and EFS in both cohorts. Conclusions: DNA methylation of key pharmacologic and leukemia-related genes is associated with clinical outcomes in pediatric AML. These findings identify biologically and clinically relevant epigenetic biomarkers that may improve risk stratification and support the development of precision medicine approaches incorporating DNA methylation profiling and epigenetic therapies in pediatric AML. Full article

Osteosarcoma (OS) is characterized by high metastatic potential and marked chemoresistance, with cancer stem cells (CSCs) serving as major drivers of malignant progression. Canine osteosarcoma (cOS) is considered an ideal comparative medicine model for human osteosarcoma (hOS). Accumulating evidence indicates that aberrant glucose metabolism and hexosamine biosynthetic pathway (HBP, hexosamine biosynthetic pathway)/O-linked N-acetylglucosamine (O-GlcNAc)ylation are involved in tumor progression; however, the precise mechanisms by which they regulate stemness in canine osteosarcoma cells remain unclear. In this study, we comprehensively employed glucose gradient culture, untargeted metabolomics, O-GlcNAc-modified proteomics, in vitro gene silencing, and a subcutaneous xenograft model in nude mice. Cellular functional assays revealed that high glucose significantly enhanced malignant phenotypes and stemness properties of canine osteosarcoma cells. Metabolomic analyses confirmed aberrant activation of the HBP in osteosarcoma cells. Further experiments demonstrated that high glucose enhances HBP flux and O-GlcNAcylation in a dose-dependent manner; silencing of glutamine-fructose-6-phosphate transaminase 1 (GFPT1), O-GlcNAc transferase (OGT), and O-GlcNAcase (OGA) verified that both the HBP pathway and O-GlcNAcylation positively regulate malignant biological behaviors and stemness maintenance. In vivo tumorigenesis assays demonstrated that OGT knockdown markedly suppressed osteosarcoma growth. O-GlcNAc-modified proteomics identified transducin-like enhancer of split 3 (TLE3), nuclear receptor corepressor 1 (NCOR1), and neurogenic locus notch homolog protein 2 (NOTCH2) as key differentially modified proteins, predominantly enriched in the Wingless/Integrated (Wnt) and Notch signaling pathways. Collectively, our findings demonstrate that high glucose activates the HBP pathway, elevates global O-GlcNAcylation levels, and modifies TLE3/NCOR1/NOTCH2, thereby promoting stemness maintenance in canine osteosarcoma stem cells. This study provides novel metabolic targets for precision therapy of osteosarcoma. Full article

WWOX and HIF1A are linked to hypoxia-driven metabolic and immune modulation in cancer. We examined whether the WWOX/HIF1A expression ratio acts as a context-dependent molecular integrator across breast cancer (BRCA) subtypes and ovarian carcinoma (OV). We analyzsed TCGA RNA-seq and clinical data from BRCA (n = 390) and OV (n = 228), using neoplasm cancer status as a proxy for disease-free survival (DFS). Patients were stratified by subtype-specific WWOX/HIF1A ratio cutpoints for exploratory Kaplan–-Meier analyses, and the ratio was also modelled as a standardizsed continuous covariate in Cox regression. Transcriptomic, pathway, immune-cell and hormone-related profiles were examined in relation to ratio status, with all multivariable and cutpoint-based findings treated as hypothesis-generating. The WWOX/HIF1A ratio does not act as a uniform or strongly predictive prognostic marker but instead delineates distinct biological states whose association with DFS is modest, context-dependent and statistically fragile in TCGA. Higher ratios are linked to more favourable DFS only in basal-like and HER2-enriched BRCA, together with oxidative phosphorylation, ribosomal and reduced ADAM10-linked Notch and monocyte/macrophage signatures. In luminal A BRCA and OV, lower ratios are linked to relatively favourable DFS and coincide with cytokine/JAK–STAT signalling, cytotoxic immune signatures and coordinated metabolic–endocrine programmes, whereas luminal B shows mixed immune and metabolic patterns. In this TCGA-based analysis, the WWOX/HIF1A ratio functions as a context-dependent molecular index of hypoxia, immune and hormone-related states rather than a strong, generalisable DFS predictor. The observed subtype-specific survival trends (high-ratio favourability in basal/HER2, low-ratio favourability in luminal A,B and OV) are non-significant and exploratory, and all ratio-based survival patterns require confirmation in independent cohorts and functional studies. Full article

Notch1 signaling is essential for endothelial cell fate determination and vascular development. While PKC has been implicated as an upstream activator of Notch1 signaling, the molecular mechanisms by which PKC promotes Notch1 proteolytic cleavage remain poorly defined. Here, using murine endothelial MS1 cells and mouse aortic endothelial cells (MAECs), we identify two cooperative pathways through which PKC drives generation of the Notch1 intracellular domain (NICD). First, PKC activation increases mRNA and protein levels of the Notch1 ligands Jag1 and Dll4. A NICD-Dll4 positive feedback loop further amplifies Notch signaling. Second, PKC upregulates hyaluronic acid synthase 2 (HAS2) mRNA and protein expression, promoting hyaluronan (HA) synthesis. Various genetic (HAS2 knockdown, CD44 knockdown, Hyal2 overexpression) and pharmacological (4-methylumbelliferone, 4-MU) treatments confirmed that HA is required for PKC to activate Notch1. Inhibition of HA production or signaling is associated with downregulation of ADAM10 on the cell surface. Direct phosphorylation of Notch1 at S1791 is dispensable for this effect. Together, these findings reveal a dual pathway model in which PKC coordinates ligand upregulation and HA signaling to promote Notch1 cleavage, identifying the extracellular matrix as a previously unrecognized regulator of Notch1 signal strength in endothelial cells. Full article