Single-cell RNA sequencing of Versican (Vcan) knockout mice after cardiac ischemia.
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ABSTRACT:
PROVIDER: PRJEB61792 | ENA |
REPOSITORIES: ENA
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Project description:The tumor derived Versican (VCAN) can be a possible mediator of the host NF-κB pathway in cancer pathogenesis. In this context, we defined VCAN function in the pathogenesis development in vitro with cell lines as well as in vivo with mouse model. We used the proteomic technique to quantify VCAN integrity and relative abundancy produced by different cancer cell lines and to link this information to our pre-existing data. These results helped us to finalise our dataset and define more clearly VCAN influence tumor development.
2023-05-10 | PXD019883 | Pride
Project description:Background: Extracellular matrix remodeling mechanisms are understudied in cardiac development and congenital heart defects. Two similar matrix-degrading metalloproteases, ADAMTS1 and ADAMTS5, are extensively co-expressed during mouse cardiac development. The mouse mutants of each have mild cardiac anomalies, but their combined genetic inactivation is precluded by tight linkage. We coupled Adamts1 inactivation with pharmacologic ADAMTS5 blockade to uncover stage-specific cooperative roles and mechanisms in mouse cardiac development. Methods: ADAMTS5 blockade was achieved in Adamts1 null mouse embryos using an activity-blocking monoclonal antibody during distinct developmental windows covering myocardial compaction or cardiac septation and outflow tract rotation. Synchrotron imaging, RNA in situ hybridization, immunofluorescence microscopy and electron microscopy were used to determine the impact on cardiac development and compared to Gpc6 and ADAMTS-cleavage resistant mouse mutants. Mass spectrometry-based N-terminomics was used to identify relevant substrates. Results: Combined inactivation of ADAMTS1 and ADAMTS5 prior to 12.5 days of gestation led to dramatic accumulation of versican-rich cardiac jelly and inhibited formation of compact and trabecular myocardium, which we also observed in mice with ADAMTS cleavage-resistant versican. Subsequently, combined knockout impaired outflow tract development and ventricular septal closure, generating a tetralogy of Fallot-like defect independently of versican proteolysis. N-terminomics of combined ADAMTS knockout and wild-type hearts identified a cleaved glypican-6 peptide only in the wild-type and showed that ADAMTS1 and ADAMTS5 each cleaved glypican-6. Paradoxically, ADAMTS1 and ADAMTS5 inactivated hearts lacked glypican-6 despite unaltered Gpc6 transcription. Gpc6-/- mice demonstrated similar rotational defects as the combined ADAMTS knockout and both had reduced Hedgehog signaling. Conclusions: ADAMTS1 and ADAMTS5 ensure proper cardiac development via cleavage of distinct proteoglycans, each with independent roles in cardiac development. Whereas versican clearance in cardiac jelly is required for proper ventricular cardiomyogenesis, glypican-6 cleavage may activate/stabilize this cell-surface proteoglycan which is required for Hedgehog signaling during outflow tract development.
2025-05-06 | PXD045706 | Pride
Project description:Background: Extracellular matrix remodeling mechanisms are understudied in cardiac development and congenital heart defects. Two similar matrix-degrading metalloproteases, ADAMTS1 and ADAMTS5, are extensively co-expressed during mouse cardiac development. The mouse mutants of each have mild cardiac anomalies, but their combined genetic inactivation is precluded by tight linkage. We coupled Adamts1 inactivation with pharmacologic ADAMTS5 blockade to uncover stage-specific cooperative roles and mechanisms in mouse cardiac development. Methods: ADAMTS5 blockade was achieved in Adamts1 null mouse embryos using an activity-blocking monoclonal antibody during distinct developmental windows covering myocardial compaction or cardiac septation and outflow tract rotation. Synchrotron imaging, RNA in situ hybridization, immunofluorescence microscopy and electron microscopy were used to determine the impact on cardiac development and compared to Gpc6 and ADAMTS-cleavage resistant mouse mutants. Mass spectrometry-based N-terminomics was used to identify relevant substrates. Results: Combined inactivation of ADAMTS1 and ADAMTS5 prior to 12.5 days of gestation led to dramatic accumulation of versican-rich cardiac jelly and inhibited formation of compact and trabecular myocardium, which we also observed in mice with ADAMTS cleavage-resistant versican. Subsequently, combined knockout impaired outflow tract development and ventricular septal closure, generating a tetralogy of Fallot-like defect independently of versican proteolysis. N-terminomics of combined ADAMTS knockout and wild-type hearts identified a cleaved glypican-6 peptide only in the wild-type and showed that ADAMTS1 and ADAMTS5 each cleaved glypican-6. Paradoxically, ADAMTS1 and ADAMTS5 inactivated hearts lacked glypican-6 despite unaltered Gpc6 transcription. Gpc6-/- mice demonstrated similar rotational defects as the combined ADAMTS knockout and both had reduced Hedgehog signaling. Conclusions: ADAMTS1 and ADAMTS5 ensure proper cardiac development via cleavage of distinct proteoglycans, each with independent roles in cardiac development. Whereas versican clearance in cardiac jelly is required for proper ventricular cardiomyogenesis, glypican-6 cleavage may activate/stabilize this cell-surface proteoglycan which is required for Hedgehog signaling during outflow tract development.
2025-05-06 | PXD045868 | Pride
Project description:Epithelial ovarian cancer (EOC) is the deadliest gynecological cancer. MicroRNAs (miRNAs) are small non-coding RNAs that play important roles in gene regulation and their dysregulation is associated with many diseases. In this study, we determined the expression and function of miR-590-3p in EOC. We found that miR-590-3p levels were higher in high-grade carcinoma when compared to low-grade or tumours with low malignant potential. Interestingly, plasma levels of miR-590-3p were significantly higher in EOC patients than in subjects with benign gynaecological disorders. Transient transfection of miR-590-3p mimics, or stable transfection of mir-590, increased cell growth, migration, and invasion. In vivo studies revealed that mir-590 accelerated tumour growth and metastasis. Using a cDNA microarray, we identified Forkhead box A2 (FOXA2) and Versican (VCAN) as a top downregulated and a top upregulated gene, respectively, by mir-590. We showed that miR-590-3p targeted FOXA2 3’ UTR to suppress its expression. In addition, knockdown of FOXA2 by siRNAs or knockout of FOXA2 by CRISPR/Cas9 enhanced cell proliferation, migration, and invasion. Overexpression of FOXA2 decreased, while knockout of FOXA2 increased, VCAN mRNA and protein levels and ChIP-qPCR revealed that FOXA2 binds to VCAN promoter. Interrogation of the TCGA ovarian cancer database revealed a negative relationship between FOXA2 and VCAN mRNA levels in EOC tumours and that high FOXA2/low VCAN mRNA levels in tumours were positively correlated with patient survival. Finally, overexpression of FOXA2 or silencing of VCAN reversed the effects of mir-590. These findings demonstrate that miR-590-3p promotes EOC development via a novel FOXA2-VCAN pathway.
2018-04-21 | GSE113440 | GEO
Project description:In this study, we directly compared turkey muscle satellite cell gene expression between satellite cells with the gene versican (VCAN) knocked down by siRNA transfection and those transfected with a lipofectamine control using our 6K Turkey Skeletal Muscle Long Oligo (TSKMLO) microarray (GPL9788). The VCAN gene was previously identified as differentially expressed by genetic line and during development of turkey skeletal muscle (Sporer et al., 2011). Gene expression changes were investigated in satellite cells after 72 h of proliferation and after 48 h of differentiation. We identified novel candidate genes and pathways as playing potentially crucial roles in the VCAN-mediated effects on the normal processes of proliferation and differentiation in turkey satellite cells previously identified by Velleman et al. (submitted).
2012-06-01 | GSE36365 | GEO
Project description:In this study, we directly compared turkey muscle satellite cell gene expression between satellite cells with the gene versican (VCAN) knocked down by siRNA transfection and those transfected with a lipofectamine control using our 6K Turkey Skeletal Muscle Long Oligo (TSKMLO) microarray (GPL9788). The VCAN gene was previously identified as differentially expressed by genetic line and during development of turkey skeletal muscle (Sporer et al., 2011). Gene expression changes were investigated in satellite cells after 72 h of proliferation and after 48 h of differentiation. We identified novel candidate genes and pathways as playing potentially crucial roles in the VCAN-mediated effects on the normal processes of proliferation and differentiation in turkey satellite cells previously identified by Velleman et al. (submitted). This experiment was designed to investigate the role of VCAN expression in turkey satellite cells during two crucial processes in muscle development: proliferation and differentiation. Satellite cells were isolated from 7-week-old turkeys from the RBC2 line. Cells were transfected with either VCAN siRNA or a lipofectamine control; VCAN expression was knocked down by over 50% with siRNA transfection (Velleman et al., submitted). Satellite cells were then induced to proliferate for 72h or differentiate for 48h; culture plates from each stage (n=4) were frozen until RNA extraction. Microarrays directly compared the VCAN-knockdown to control from each of 4 culture plates and utilized a dye swap to equal 8 arrays for each of the cell developmental stages, proliferation and differentiation, and 16 arrays for the overall experiment investigating the role of VCAN expression in satellite cell development and function. Hybridizations were performed in random order.
2012-05-31 | E-GEOD-36365 | biostudies-arrayexpress
Project description:Abnormal cardiac trabeculation often leads to congenital heart disease (CHD). TIE2-ANG1 signaling is a key regulator of endocardial growth during trabeculation. However, little is known about the underlying mechanisms. Here we demonstrate that loss of endocardial Tie2 via Nfatc1Cre resulted in increased nuclear localization of Forkhead box O1 (FOXO1), reduced phosphorylation of AKT (the direct regulator of FOXO1) and reduced expression of several proliferation promoting genes in endocardial cells (EdCs). The cardiac defects seen in endocardial Tie2-deficient hearts were phenocopied by forced activation of FOXO1 within the endocardium and partially rescued by loss of FoxO1. Conversely, excessive PI3K signaling (the direct regulator of phosphorylation of AKT) in EdCs caused cardiac hypotrabeculation. Further, we identify that endocardial loss of Tie2 resulted in impaired cleavage of versican, which was associated with enhanced expression of ADAMTS endogenous inhibitor TIMP3 in Tie2-cko EdCs. Thus, our results suggest that TIE2 signaling in the early embryonic endocardium promotes endocardial growth via sequestering FOXO1 within the cytoplasm and modulates cardiomyocyte proliferation via stimulating versican cleavage to orchestrate myocardial trabeculation, providing insights into the pathogenesis of CHD.
2026-06-30 | GSE334972 | GEO
Project description:We generated a tamoxifen-inducible, adipocyte-specific ATGL-KO animals to analyze the role of adipocyte ATGL in cardiac ischemia/reperfusion. KO was induced in 10 weeks old male mice via 7d of consecutive i.p.-injections of 500µg hydroxytamoxifen/day. Pnpla2-flox homozygous, Cre-negative littermates were used as control. After 2 weeks washout phase mice underwent closed chest ischemia with first ligature-induction surgery and 3-7 d later 1 hour closed chest ischemia followed by 24 h reperfusion. Mice were sacrificed and hearts excised to perform spatial transcriptomic analysis on 10X Visium Gene Expression slides. The goal was to identify differences in gene expression between KO and control in the different zones of the heart after ischemia (eg. ischemic, border and remote zone).
2026-02-28 | E-MTAB-12700 | biostudies-arrayexpress
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Project description:The activation and accumulation of lung fibroblasts, leading to excessive extracellular matrix (ECM) deposition, is a pathogenic hallmark of Idiopathic Pulmonary Fibrosis (IPF), a lethal and currently incurable disease. The collagen-rich fibrotic ECM perpetuates fibroblast activation and accumulation, which includes the promotion of podosome formation and ECM invasion. Proteoglycans (PGs), a significant component of the interstitial ECM, fine-tune the overall tissue architecture determined by fibrous proteins and regulate several ECM-controlled signalling pathways. In this report, increased expression of Versican (VCAN ), a multifunctional PG, was detected in human and mouse pulmonary fibrosis, predominantly in monocytic cells and fibroblasts. Genetic reduction of Vcan expression in mice promoted collagen expression and modulated pulmonary ECM composition and structure, exacerbating pulmonary fibrosis and delaying its resolution. Reduced ECM Vcan levels resulted in longer, thicker, and more tangled collagen fibrils, which stimulated podosome formation in fibroblasts and their ECM invasion. Moreover, the decrease of Vcan in the ECM and the ensuing reorganisation stimulated Tenascin-C (TNC) expression from fibroblasts, which was further shown to be a potent, TLR4-dependent, autologous podosome inducer, promoting ECM invasion. Thus, Vcan expression from fibroblasts serves as an autologous fibrotic brake, controlling the underlying ECM composition, structure and mechanotransduction, and suppressing fibroblast invasion and pulmonary fibrosis.
2026-02-23 | PXD063546 | Pride