PRJNA497133
miRNA-Sequencing data of extracellular vesicles derived from human induced pluripotent stem cells (iPSC-EVs)
Progression of fibrosis and the development of cirrhosis are responsible for the liver related morbidity and mortality associated with chronic liver diseases. There is currently a great unmet need for effective anti-fibrotic strategies. Stem cells play a central role in wound healing responses to restore liver homeostasis following injury. Here we tested the hypothesis that extracellular vesicles isolated from induced pluripotent stem cells (iPSC) modulate stellate cell activation and may have anti-fibrotic effects. Human iPSCs were generated by reprogramming primary fibroblasts isolated from human skin biopsies. EVs were isolated by differential centrifugation and quantified by flow cytometry (FACS) and characterized by dynamic light scattering (DLS) and electron microscopy (TEM). Primary human hepatic stellate cells (HSC) were isolated from liver donors and primed with TGF-ß (10ng/mL) with or without iPSC-EVs. The role and efficacy of iPSC-EVs on HSC phenotype and liver fibrosis was assessed in vitro and in vivo in a murine model of liver fibrosis. Characterization of iPSC-derived EVs identified two subpopulations of EVs: small (10-30 nm) and large vesicles (100-1,000 nm). EVs were also detected by electron microscopy and their quantification by FACS showed that iPSC released large amounts of EVs. Fluorescent tracing assay detected iPSC-EVs internalized into HSC. Further analysis of the HSC responses occurring during fibrogenesis showed that iPSC-derived EVs reduced HSC chemotaxis and proliferation. miRNA-sequencing of iPSC-EVs revealed the presence 22 highly expressed miRNAs, among which and miR-92a-3p resulted the most expressed. Coding transcriptome analysis identified 60 genes down-modulated and 235 up-regulated when comparing in HSC primed with TGF-ß and exposed to iPSC-EVs versus not exposed cells. A biweekly tail vein injection of 1x106 murine iPSC-EVs in mice exposed to carbon tetrachloride (CCl4) for 3 weeks, showed significant reduction in liver fibrosis and HSC activation. Conclusions: Results of this study identify iPSC-EVs as a novel anti-fibrotic approach that may reduce or reverse liver fibrosis in patients with chronic liver disease.