Project description:CADM1, an immunoglobulin superfamily member, frequently inactivated but function as a tumor suppressor in many solid tumors. However, how CADM1 expression in ovarian cancer cells and the mechanisms of its tumor suppressor function is not fully understood. We created ovarian cancer ES-2 cell lines in which CADM1 was stably up-regulated. Genes differentially expressed in CADM1-overexpressing ES-2 cells.
Project description:Background and aims: Cytotoxic T cells have long been postulated to facilitate autoimmune destruction of intestinal epithelium; and the precise causal events leading to inflammatory bowel diseases (IBDs) remain unresolved. CADM1, a membrane adhesion protein which can shed its extracellular ectodomain enters the systemic circulation as well as bind the receptor CRTAM present on CD8+ T cells. Methods: We performed RNA and small RNA sequencing on colon tissue from IBD and non-IBD (NIBD) control patients. We performed spatial transcriptomics and stimulated lamina propria mononuclear cells (LPMCs) with the soluble form of CADM (sCADM1) in these patients. Dextran Sulfate Sodium (DSS) was used to induced colitis in a conditional myeloid Cadm1 loss-of-function mouse. Results: MicroRNA-375 was significantly decreased while its direct target CADM1 was markedly up-regulated in UC patients compared to NIBD control subjects. Single cell proteomics data identified CADM1 enrichment in multiple clusters including macrophages and dendritic cells from colonic tissue of human subjects with IBD. An increased number of CADM1+CD68+ cells was detected adjacent to CD8+ T-cells within the intestinal epithelium of colon sections from patients with ulcerative colitis (UC) compared to NIBD subjects. Myeloid Cadm1 conditional knockout mice were protected against DSS-induced colitis indicating Cadm1 may facilitate binding/localization of cytotoxic cell populations within the gut epithelium. Furthermore, we observed that serum levels of the cleaved extracellular ectodomain of CADM1 (sCADM1) are elevated in medically refractory UC patients compared to non-IBD and UC donors in remission and treatment of LPMCs with recombinant sCADM1 enhanced STAT3 phosphorylation. Conclusions: Together these results identify CADM1 as a potent mediator of pro-inflammatory signaling pathways and demonstrate its potential as a diagnostic and therapeutic target for preventing the IBDs.
Project description:CADM1 expression in non-adherent conditions was found to potently induce caspase-independent cell death. We sought to use microarray analysis to glean insights to the mechanism associated with CADM1 induced non-adherent cell death.
Project description:Cytotoxic T cells have been postulated to facilitate the destruction of intestinal epithelium in inflammatory bowel diseases (IBDs). CADM1, which encodes a membrane adhesion protein that can bind the T cell receptor CRTAM, was markedly up regulated in colon of IBD patients compared to non-IBD (NIBD) patients. We then identified CADM1 enrichment in multiple immune cell clusters including macrophages and dendritic cells in the colons of IBD patients. Increased numbers of CADM1+ myeloid cells were measured adjacent to CD8+ T cells within colons of ulcerative colitis patients compared to NIBD patients. Conditional deletion of Cadm1 in myeloid cells resulted in reduced numbers of activated T cell populations and protected mice from chemical-induced colitis. Similarly, administration of a Cadm1 ‘neutralizing’ antibody which binds its extracellular domain reduced tissue inflammation and breakdown of the intestinal epithelium and crypts after induction of colitis in mice. Lastly, serum levels of sCADM1 were elevated in IBD patients compared to NIBD controls and treatment of LPMCs with recombinant sCADM1 enhanced inflammatory STAT3 phosphorylation. Therefore, we concluded that CADM1 is a mediator of pro-inflammatory signaling cascades in the colon and a potential therapeutic target for the IBDs.
Project description:Overexpression of transcription factor Sox17 in human ES cells-derived endothelial cells and hematopoietic cells enhances expansion of hemogenic endothelium-like cells.
Project description:RNA-seq of murine ES cells with Lima1 knockout, murine EpiS cells with Lima1 overexpression and human iPS cells with LIMA1 overexpression
Project description:In chimera assays, murine naïve embryonic stem (ES) cells usually produce better chimeras than the more primed epiblast-derived stem (EpiS) cells. Overexpression of the cytoskeleton-associated protein LIMA1/EPLIN in EpiS cells improves the rate of successful chimeras, while the knockout of LIMA1 in ES cells results in a metabolic state reminiscent of EpiS cells. To investigate the effects of LIMA1 we performed RNA-seq in Lima1 loss-of-function murine ES cells and in Lima1 gain-of-function murine EpiS cells and human induced pluripotent stem cells.
Project description:miRNA abnormalities are increasingly relevent to cancer development, We used microarrays to detail the global programme of gene expression upon miR-483 overexpression in sarcoma cell line MHH-ES-1.
Project description:Empty vector, SRF-dM-VP16 (control construct lacking the SRF DNA binding domain), or SRF-VP16 were expressed in wild-type ES cells (E14wt), SRF heterozygotes (E99-/+) or two SRF ko cell lines (E81-/-,E100-/-). RNA was harvested 72h after transfection. Each condition was performed in duplicate, except E100-/- vector transfected. Keywords: ordered
