Paracrine effect mediated by extracellular vesicles derived from IGF-I overexpressing human umbilical cord perivascular cells in a murine model of experimental liver fibrosis

DOMÍNGUEZ L ; BAYO J; MALVICINI M; ATORRASAGASTI C; RODRIGUEZ M; ONORATO A; CANTERO M; GARCIA M; YANARELLI G; FIORE E; MAZZOLINI G

Mar del Plata

Congreso; Reunión Anual de Sociedades de Biociencia (SAIC SAFE SAB SAP); 2019

Cirrhosis is the result of chronic liver damage/regeneration cycles and fibrosis accumulation. Human umbilical cord perivascular cells (HUCPVC) are mesenchymal stromal cells that could allow tissue regeneration by secretion of soluble factors and extracellular vesicles (EV). We previously demonstrated that HUCPVC engineered to produce Insulin Growth Factor like-I (IGF-I-HUCPVC) ameliorate liver fibrosis in mice. Our aim is to evaluate the role of EVs in the therapeutic effect of IGF-I-HUCPVC in liver fibrosis.Conditioned media (CM) or EV depleted CM (DCM) were collected from HUCPVC infected with adenovirus codifying with IGF-I or green fluorescence protein (GFP). EVs were isolated from CM by differential centrifugation and characterized by electron microscopy, dynamic light scattering and flow cytometry to test shape, size and EV markers expression respectively. IGF-I levels were assayed in EV after lysis and/or dialysis by ELISA. Fibrosis was induced in BALB/c mice by administration of thioacetamide for 8 weeks (600 mg/kg/week). On week 6, IGF-I-HUCPVC and GFP-HUCPVC derived EV, CM or DCM were intravenously administrated (3 doses, 15ug/dose/mice, every 5 days) and at week 8 liver samples were collected. Hepatic Stellate Cells (CFSC-G2 cell line) and hepatic macrophages (Mø) were incubated in vitro with EV, CM or DCM, and gene expression evaluated by qPCR.CFSC-G2 incubation with CM or EV derived from IGF-I-HUCPV downregulates the expression of COL1A2 and a-SMA in comparison with DCM-IGF-I-HUCPVC (p