Search Results (453)

Syntaphilin (SNPH), initially considered a neuron-specific protein, has recently been found to be widely expressed across various cancers. Mechanistically, SNPH inhibits mitochondrial transport to the cortical cytoskeleton, thereby suppressing cancer cell migration and metastasis. Although SNPH is known to participate in the metastatic progression of multiple malignancies, its precise underlying mechanism remains obscure. In this study, we investigated the role of SNPH in the epithelial–mesenchymal transition (EMT) and invasiveness of gastric cancer cells. Knockdown of SNPH in SNU-638 gastric cancer cells significantly enhanced their migratory and invasive capacities by approximately 1.4-fold and 2.5-fold, respectively. This knockdown concurrently increased focal adhesion kinase (FAK) phosphorylation, upregulated the EMT marker vimentin, and increased the expression of key EMT-related transcription factors, including Snail, Slug, and Twist. Furthermore, SNPH depletion induced the phosphorylation of nuclear factor kappa B (NF-κB), a transcription factor regulating matrix metalloproteinase-9 (MMP-9), which subsequently upregulated MMP-9 mRNA expression. This cascade promotes extracellular matrix degradation, thereby increasing metastatic potential. Notably, these phenotypic and molecular changes were completely reversed upon SNPH overexpression in SNU-638 cells. Taken together, our results demonstrate that SNPH deficiency enhances the migration and metastatic potential of gastric cancer cells by driving EMT and invasion via the FAK/NF-κB/MMP-9 signaling pathway. Consequently, we propose that SNPH represents a novel biomarker and a promising therapeutic target for mitigating gastric cancer metastasis. Full article

The adult mammalian heart exhibits limited regenerative capacity. Although the epicardium contributes to cardiac injury responses and remodeling, expandable adult human in vitro models for investigating epicardial activation and epithelial-to-mesenchymal (EMT)-associated cellular responses remain limited. Here, we isolated human epicardium-derived cells from the adult heart auricle, expressing WT1⁺/MSLN⁺/CRIP1⁺ and generated an expandable immortalized epicardium-derived cell (iEPDC) population, allowing the investigation of intercellular dynamics upon EMT activation. TGFβ signaling was modulated using SB431542 or TGFβ3. Morphological, immunocytochemical, transcriptomic and functional analyses were performed to investigate treatment-dependent responses. SB431542-treated iEPDCs displayed epithelial-like characteristics and elevated WT1, MSLN and CRIP1 expression, with CRIP1 detected at both transcript and protein levels. TGFβ3-treated cells expressed the mesenchymal markers VIM and CD105 and exhibited spindle-shaped morphology, increased migratory behavior and upregulation of mesenchymal- and remodeling-associated markers. Transcriptomic analyses revealed distinct treatment-dependent profiles, enrichment of ‘focal adhesion’, ‘ECM-receptor interaction’ and cytoskeleton-associated pathways in TGFβ3-treated iEPDCs and intermediate transcriptional characteristics in untreated cells. Together, these findings establish adult iEPDCs as an expandable in vitro model for investigating TGFβ-dependent epicardial activation and EMT-associated processes in the adult human heart. Furthermore, the integration of phenotypic, transcriptomic and functional findings revealed the treatment-responsive plasticity of adult human iEPDCs, supporting future studies of injury-associated epicardial activation. Full article

Osteoporosis (OP) and osteosarcoma (OS) are biologically distinct skeletal disorders that share dysregulated bone remodeling, inflammatory signaling, and microenvironmental interactions. This study performed an integrative transcriptomic analysis to identify shared osteoimmune regulatory programs in circulating monocytes from postmenopausal women with OP and OS tumors from Central Mexican cohorts. Two independent RNA-seq cohorts were analyzed separately and then integrated: circulating monocytes from postmenopausal women with OP and controls (7 OP and 7 controls), and donor-matched OS tissues (7 tumors and 7 healthy bone samples). Differential expression, module-based filtering, pathway enrichment, cross-cohort functional integration, directional concordance, and targeted interaction network analyses were performed. The OP cohort showed 169 differentially expressed genes, whereas the OS cohort showed 2135 genes. Module-based filtering retained 82 genes in OP and 278 in OS, with only six genes directly shared. However, pathway-level integration identified convergent signals involving PI3K-Akt, HIF-1 signaling, lipid and atherosclerosis, phagosome, endoplasmic reticulum protein processing, focal adhesion, proteoglycans in cancer, and cancer-related pathways. Directional analysis of 27 shared pathway-associated genes revealed predominantly discordant regulation, with CTNNB1 emerging as a central network node. These findings suggest that OP and OS converge through specific osteoimmune pathways rather than through a uniform shared gene program. Full article

The gastrointestinal epithelium forms a critical barrier that regulates nutrient absorption while preventing the translocation of harmful luminal contents. Disruption of this barrier initiates a coordinated wound healing response involving epithelial restitution, proliferation, and differentiation. Focal adhesion kinase (FAK) is central to this process, regulating focal adhesion (FA) turnover and cytoskeletal dynamics required for epithelial migration. Activation of FAK via phosphorylation at tyrosine 397 (Y397) promotes cell motility, proliferation, and survival, whereas loss of function impairs mucosal repair and exacerbates tissue injury. Effective wound healing also requires tight regulation of reactive oxygen species (ROS). The nuclear factor erythroid 2-related factor 2 (NRF2) pathway governs antioxidant defenses by inducing cytoprotective genes, including SOD1, CAT, GCLC, GCLM, and NQO1, restoring redox homeostasis and limiting inflammation. NRF2 deficiency results in increased oxidative stress, heightened inflammatory signaling, and delayed wound healing. While both FAK and NRF2 are independently essential for gastrointestinal wound healing, their mechanistic relationship remains unclear. Emerging evidence suggests that they may be functionally linked through redox-dependent signaling where FAK-mediated ROS production may promote NRF2 activation, while NRF2-driven antioxidant responses maintain conditions necessary for sustained FAK signaling. This coordinated interaction highlights a redox-sensitive feedback mechanism critical for efficient gastrointestinal wound repair. Full article

Intramuscular fat content is a key determinant of pork quality, influencing traits such as tenderness, juiciness, and flavor. However, the molecular mechanisms regulating intramuscular fat deposition in indigenous pig breeds remain incompletely understood. This study aimed to identify genes and regulatory mechanisms associated with intramuscular fat accumulation in Hezuo pigs. Longissimus dorsi muscle samples from Hezuo pigs with extreme high and low intramuscular fat contents were subjected to chromatin accessibility profiling and transcriptome sequencing. Comparative analyses identified 2201 differentially accessible chromatin regions and 588 differentially expressed genes between the two groups. Functional enrichment analyses indicated that these genes were mainly involved in lipid metabolism, focal adhesion, extracellular matrix–receptor interaction, fatty acid metabolism, and adenosine monophosphate-activated protein kinase signaling. Integration of chromatin accessibility and gene expression datasets identified 92 co-regulated genes associated with intramuscular fat deposition, including MYLK3, PDGFC, PAK1, IGF1R, LAMA4, DIAPH1, SDC4, GADD45G, and RXRG. These findings reveal regulatory networks underlying intramuscular fat accumulation in Hezuo pigs and provide candidate genes and molecular resources for improving meat quality through genetic selection and breeding programs. Full article

Dextran sodium sulfate (DSS) was used to induce ulcerative colitis in a porcine model to investigate the transcriptional responses in inflamed colonic tissue. Eleven pigs were divided into two groups, of which five pigs were administered an oral dose of DSS daily for five days, whereas six non-treated pigs served as a control group. Differences in transcript expression between treated and control pigs identified 425 down-regulated and 780 up-regulated mRNAs in response to DSS treatment. Fifty-nine differentially expressed miRNAs were also identified, comprising 20 up-regulated and 39 down-regulated miRNAs. The top enrichment KEGG pathways for the up-regulated genes were breast cancer (ssc05224), gastric cancer (ssc05226), focal adhesion (ssc04510), and the PI3K-Akt signaling pathway (ssc04151). The top gene ontology terms for the up-regulated genes were blood vessel development (GO:0001568), extracellular matrix and external encapsulating structure (GO:0031012). Protein–protein interaction network analysis identified three hub genes, including LOXL1, MFAP2, and FSTL3. Seventeen high-confidence miRNA-mRNA pairs were recognized, and two genes (CD101 and AVL9) have been confirmed as targets for ssc-miR-24-3p by dual-luciferase reporter assay. Our study provides a better understanding of the key roles of mRNAs and miRNAs in regulating DSS-induced colitis in pigs and defines sets of coding and non-coding RNA transcripts, which may serve as intervention targets or biomarkers for ulcerative colitis. Full article

Almond-derived peptide fractions (APs) represent a promising yet underexplored class of plant-based bioactive compounds for ulcerative colitis (UC) management. This study investigated the protective effects of APs against dextran sulfate sodium (DSS)-induced colitis in male BALB/c mice using integrated 16S rRNA sequencing and transcriptomic analysis. Mice received low-dose (200 mg/kg) or high-dose (400 mg/kg) APs during 7-day DSS administration. Both doses significantly attenuated body weight loss, disease activity index, colon shortening, and histopathological damage. Compared with the DSS group, low-dose and high-dose APs reduced disease activity index by 65.9% and 52.4%, increased colon length by 30.0% and 26.5%, decreased histopathological scores by 39.6% and 55.1%, and lowered colonic myeloperoxidase activity by 13.9% and 34.1%, respectively. APs also modulated inflammatory cytokines, improved mucus-barrier-related indices, and enhanced continuous ZO-1 localization at epithelial junctions. Additionally, 16S rRNA sequencing indicated that AP treatment was associated with altered genus-level microbiota profiles after DSS exposure, including changes in Ligilactobacillus, Escherichia-Shigella, and Parabacteroides-related taxa. Transcriptomic analysis suggested dose-associated response patterns: low-dose APs were associated with broader immune-related transcriptional changes, whereas high-dose APs were more closely associated with extracellular matrix organization, focal adhesion, and genes enriched in the PI3K-Akt signaling pathway. These findings support the protective effects of APs in DSS-induced acute colitis and suggest potential associations among microbiota alterations, host transcriptional responses, and mucosal repair. Full article

Primordial germ cells (PGCs) are the earliest precursors of gametes and essential cellular materials for poultry germplasm conservation and genetic modification. In this study, PGCs isolated from 2.5-day male and female Silkie chicken embryos were defined as circulating PGCs (cPGC_M and cPGC_F), whereas PGCs isolated from 8.5-day male and female embryos were defined as gonad-derived PGCs (gPGC_M and gPGC_F). RNA sequencing with three biological replicates per group was performed to characterize developmental-stage- and sex-associated transcriptional programs. Comparative transcriptomic analyses identified stage-associated differences in pathways related to cell–matrix interaction, focal adhesion, PI3K–Akt signaling, and stem cell pluripotency, with more differentially expressed genes detected in the female than in the male stage comparison. Sex-biased expression was detectable at the circulating stage, mainly reflecting W- and Z-linked expression differences, and the number of sex-biased genes was greater after gonadal colonization, primarily because more autosomal differentially expressed genes were detected. Candidate genes and pathways, including HINTW, DMRT1, SOX2, EDNRB, LPAR4, GFRA3, ECM–receptor interaction, ribosome-related modules, and calcium signaling, were associated with these comparisons and are presented as targets for future validation rather than as confirmed regulators. Because the study used three biological replicates per group, RT-qPCR corroboration was limited to six female stage-associated genes, and no functional perturbation assays were performed, the findings are descriptive and do not establish causal mechanisms of PGC migration, gonadal colonization, or sex differentiation. Within these limitations, the dataset provides an exploratory resource for subsequent studies of chicken PGC biology and biotechnology. Full article

The small heat shock protein HSPB1 is a ubiquitously expressed mechanoresponsive chaperone essential for cytoskeletal remodeling under mechanical load. Mutations in HSPB1, including S135F, cause Charcot-Marie-Tooth (CMT) peripheral neuropathy, yet the mechanisms underlying the selective vulnerability of peripheral nerves remain enigmatic. Here we demonstrate that substrate stiffness is a critical determinant of HSPB1^S135F-mediated neurodegeneration. Using stiffness-tunable polydimethylsiloxane (PDMS) substrates (1 kPa, 10 kPa, 2 MPa) and uniaxial cyclic stretch, we show that primary dorsal root ganglia (DRG) neurons and SH-SY5Y cells expressing HSPB1^S135F exhibit profound deficits in mechanoadaptation. On compliant substrates (10 kPa), HSPB1^S135F causes stretch-induced axon fragmentation and neuronal death, whereas HSPB1^WT confers robust neuroprotection. HSPB1^S135F also disrupts stiffness-directed neuritogenesis in differentiated SH-SY5Y cells: HSPB1^WT-expressing cells show optimal axonal outgrowth and βIII-tubulin expression on 10 kPa substrates mimicking muscle tissue stiffness, while HSPB1^S135F mutants display disorganized focal adhesions and complete differentiation failure. Mechanistically, we uncover that HSPB1^S135F dysregulates stage-specific transglutaminase (TGase) expression—insufficient TGase during early neuritogenesis impairs filopodia stabilization, whereas aberrant TGase persistence at late stages constrains axon extension. Our findings establish HSPB1 as a biomechanical sensor that integrates ECM stiffness signals to coordinate peripheral nerve regeneration, and identify defective mechanoadaptation as a previously unrecognized pathomechanism in CMT. These results open new avenues for stiffness-targeted therapeutic strategies in peripheral neuropathy. Full article

Glioblastoma (GBM) remains a lethal brain tumor with poor prognosis and limited therapeutic efficacy from temozolomide (TMZ) treatment. Prodigiosin (PG), a bioactive secondary metabolite, has demonstrated anti-tumor activity across a broad spectrum of tumors. This study aims to investigate the therapeutic potential and mechanism of PG combined with TMZ in treating GBM. The results demonstrated that the combination of PG and TMZ synergistically inhibited GBM cell proliferation, triggered apoptosis, and suppressed migration and invasion. Transcriptomic analysis revealed downregulation of focal adhesion and related signaling pathways. Functionally, the combination therapy reduced focal adhesion numbers and AKT phosphorylation. Co-treatment with PG and TMZ impaired autophagic flux, evidenced by LC3-II and p62 accumulation. Furthermore, the anti-proliferative effect and the accumulation of LC3-II and P62 by the combination therapy were enhanced by the autophagy inhibitor chloroquine (CQ) but not reversed by the autophagy activator rapamycin (Rapa), confirming autophagy inhibition as a key mechanism. In conclusion, PG sensitized GBM cells to TMZ by impairing autophagy and focal adhesion signaling, providing a preclinical rationale for the combinatorial strategy. Full article

Junken meat sheep exhibit a characteristic postnatal coat-color transition, in which the initially black fleece gradually fades and develops into a white-trunk phenotype; however, the transcriptional basis of this developmental change in follicular pigment output remains unclear. In this study, three Junken meat sheep lambs showing a natural postnatal black-to-white coat-color transition were sampled longitudinally at the newborn black-fleece stage and the 179-day white-trunk stage, generating three matched biological pairs for RNA-seq analysis. Representative candidate genes were further validated by RT-qPCR. Differential expression analysis identified 1657, 400, and 1086 differentially expressed genes in the C11 vs. C1, C22 vs. C2, and C33 vs. C3 comparisons, respectively. Functional enrichment analysis indicated that these genes were mainly associated with tyrosine metabolism, ECM–receptor interaction, focal adhesion, Phosphoinositide 3-kinase-Akt signaling pathway (PI3K-Akt), arachidonic acid metabolism, estrogen signaling, and immune-related pathways. Integrated analysis of shared downregulated genes and expression patterns highlighted candidate genes related to pigmentation, the ECM/follicular microenvironment, and regulatory or metabolic processes. Pigmentation-related genes, including SOX10, TYR, TYRP1, PMEL, OCA2 and SLC45A2, were generally downregulated in 179-day white-trunk-stage skin, while changes in ECM- and metabolism-related genes suggested altered follicular microenvironmental regulation. These findings identify candidate transcriptional features associated with developmental coat-color fading in Junken meat sheep and support a follicular microenvironment–melanogenesis expression axis as a transcriptome-based framework for further investigation. Full article

Aspartame is a widely used artificial sweetener, but its possible relationship with rheumatoid arthritis (RA) remains insufficiently understood. This study aimed to explore, rather than prove, potential molecular links between aspartame-related targets and RA-associated gene networks. Three public RA transcriptomic datasets (GSE55235, GSE55457, and GSE77298) from the Gene Expression Omnibus (GEO) database were integrated as discovery/training data. Because these datasets included different tissue origins, batch correction was used to reduce dataset-level technical variation, whereas tissue-origin-related biological variation was not assumed to be fully removable. After differential expression analysis, RA-associated differentially expressed genes (DEGs) were identified. The single-cell dataset GSE200815 was used for cell annotation and cellular expression visualization; because its comparator group consists of psoriatic arthritis (PsA) samples rather than healthy controls, single-cell results were interpreted as RA-vs-PsA observations and were not treated as disease-versus-healthy-control evidence. Potential targets of aspartame were retrieved from ChEMBL, SwissTargetPrediction, and the Similarity Ensemble Approach (SEA), and were intersected with RA-related DEGs to construct an aspartame-gene-RA regulatory network. Diagnostic models were developed using 113 machine-learning algorithm combinations to determine an optimal multigene model and its core genes. HLA-DRB1 was selected for exploratory scTenifoldKnk-based virtual knockout mainly because it was included in the optimal model and has a well-established role in RA immunogenetics; the single-cell analysis was used only to describe cellular distribution in the RA/PsA dataset. Molecular docking was then used to evaluate the possible interaction between aspartame and HLA-DRB1. Forty-four intersected genes linked the predicted aspartame targets with RA DEGs. The random forest plus partial least-squares generalized linear model (RF + plsRglm) identified 16 core genes. Network-level interpretation indicated that these genes were distributed across immune/antigen-processing, inflammatory-signaling, protease/extracellular-matrix-remodeling, adhesion, metabolic, and proliferation-related modules; therefore, HLA-DRB1 was treated as a prioritized immune-module candidate rather than as the sole driver of the network. Following virtual knockout of HLA-DRB1, affected genes were enriched in extracellular matrix organization, extracellular structure organization, extracellular matrix, collagen trimer, extracellular matrix structural constituent, and collagen binding. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways included integrin signaling, focal adhesion, proteoglycans in cancer, cytoskeleton in muscle, and phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) signaling. Molecular docking showed a minimum binding energy of −6.7 kcal/mol, which was more negative than the preset stability criterion of −5.0 kcal/mol, and the docking pose suggested contacts around ARG-146. This integrative analysis suggests a hypothesis-generating association between aspartame-related predicted targets and RA-relevant molecular networks involving HLA-DRB1 and other core genes. The findings do not establish causality and require experimental, epidemiological, biophysical, and tissue-stratified validation before any causal or clinical inference can be made. Full article

Endothelial dysfunction is an early event in the development of cardiovascular diseases and is characterized by impaired barrier function, inflammatory activation of endothelial cells (ECs), and alterations in lipid metabolism. In addition to typical (mononuclear) endothelial cells (TECs), multinucleated variant endothelial cells (MVECs) are present within the vascular wall; however, their functional role remains poorly understood. The aim of the present study was to investigate the molecular and functional characteristics of MVECs and their potential contribution to the development of endothelial dysfunction. Primary human umbilical vein endothelial cells (HUVECs) were used, and multinucleated cells were generated by polyethylene glycol-induced fusion. Cells were incubated under control conditions or exposed to low-density lipoproteins (LDL; 100 µg/mL, 24 h). A comprehensive analysis was performed, including transcriptomic and proteomic (secretome) profiling using gene set enrichment analysis (GSEA), as well as functional assays assessing transendothelial LDL transport, intracellular cholesterol accumulation, macrophage migration, and the expression and secretion of pro-inflammatory cytokines (IL-6, IL-8). MVECs exhibited pronounced differences compared to TECs. GSEA revealed reduced enrichment of pathways related to canonical nuclear factor kappa B (NF-κB) signaling and negative regulation of NF-κB transcription factor activity, actin cytoskeleton organization, focal adhesion assembly, basement membrane organization, and vesicle-mediated transport in MVECs relative to TECs, indicating impaired cytoskeletal integrity, altered cell–matrix interactions, dysregulated inflammatory signaling, and reduced vesicular trafficking activity. Functionally, MVECs demonstrated an increased capacity for cholesterol accumulation and enhanced transendothelial migration of macrophages. Notably, transendothelial LDL transport across the MVEC monolayer was not increased, suggesting a predominance of intracellular lipid accumulation. MVECs also exhibited a pronounced pro-inflammatory phenotype, characterized by elevated expression and secretion of IL-6 and IL-8. Taken together, these findings indicate that MVECs represent a functionally altered endothelial phenotype with impaired barrier function, dysregulated lipid metabolism, and enhanced inflammatory activity. Local accumulation of MVECs within the vascular wall may contribute to the formation of pro-atherogenic regions and play a role in the initiation and progression of endothelial dysfunction. Full article

Vibrio tubiashii infection has led to several Babylonia areolata pandemics on the southeast coast of China, yet the immune response of the ivory shell against V. tubiashii and the specific pathogen–host interaction remain unclear. This dynamic study aimed to characterize the response of B. areolata to V. tubiashii infection with the use of pathology, transcriptomics, an enzymatic assay, and inflammatory cytokines. Hepatopancreatic cells showed marked vacuolar degeneration with intact cell membrane and extensive cytoplasmic vacuolization after infection. The dynamic transcriptome of the hepatopancreatic tissue was analyzed by RNA-seq after V. tubiashii infection, and a total of 2733 (3 h), 5610 (24 h), 3323 (48 h), and 418 (72 h) differentially expressed genes (DEGs) were identified during infection. The GO and KEGG analyses showed that the DEGs were enriched in metabolic regulation, lysosome, and multiple immune-related pathways such as the MAPK signaling pathway. The immune response of B. areolata was distinct, where the early stage of immune response (3 h) showed binding, focal adhesion, and apoptosis, as well as an activated antioxidant system. Here, expression of TNF-α, IL-1, and IL-8 was significantly increased in the hepatopancreas, whereas expression of IL-6 and IL-17 increased afterward. During the middle stage (24 h and 48 h), a large number of DEGs were suppressed, especially those associated with metabolism and lysosomes, although their expression returned to normal during prolonged infection (72 h). The PPI network showed that ppp2, atp6, and sos1 were the top immune-related DEGs during infection. Key infection-related and time-course-related genes were analyzed by WGCNA. This study illustrates that oxidative stress, inflammation, and apoptosis are strategies of the hepatopancreatic immune response in B. areolata against V. tubiashii infection and enlightens conservation and production by furthering our understanding of gastropod immunity. Full article

Plasma exosomes have been identified to be involved in embryo development and implantation. In this study, the plasma exosomes of seven pregnant cows on day 16 and seven non-pregnant cows were collected. Exosome miRNA profiles were revealed by RNA sequencing technology and the abundance of exosome miRNA between them were compared. Seven miRNAs were significantly differently expressed (p-value < 0.05, |log2(FoldChange)| > 1), with five miRNAs upregulated and two downregulated in the pregnant cow group. The most significant miRNA was bta-miR-136 (downregulated in pregnant cows, p-value < 0.005). A total of 5828 target genes were computationally predicted for the seven miRNAs. GO and KEGG enrichment analyses showed these targets were significantly involved in several biological processes and pathways associated with embryo implantation and endometrial development, such as cell adhesion, cell junction, focal adhesion, Ras signaling pathway and Rap1 signaling pathway. This study identifies seven differentially expressed plasma exosomes miRNAs on day 16 of pregnancy in cows, with bta-miR-136, bta-miR-199b, and bta-miR-382-3p emerging as candidate miRNAs for future studies on maternal–fetal communication and embryo implantation. Full article