EXTRACELLULAR VESICLES DERIVED FROM MESENCHYMAL STROMAL CELLS: DELIVERY OF THERAPEUTICS GENES FOR LIVER CIRRHOSIS

ESTEBAN FIORE; DOMÍNGUEZ, LUCIANA; MILAGROS ALBORNOZ; JUAN BAYO; CANTERO, MARIA JOSE; BARBARA BUELONI; CATALINA ATORRASAGASTI; MARIANA GARCIA; GUSTAVO YANNARELLI; GUILLERMO MAZZOLINI

Congreso; Reunión Anual 2022 de la sociedad argentina de fisiologia; 2022

Liver cirrhosis involves chronic damage, wound healingand fibrogenic processes. Mesenchymal stem cells(MSC)-derived extracellular vesicles (EVs) are an interestingtherapeutic option for regenerative medicine. Wepreviously demonstrated that EVs mediates the therapeuticeffect of human umbilical cord perivascular cells (atype of MSC) over-expressing IGF1 (AdIGF-I-HUCPVC)on liver fibrosis. Our aim is to establish a new tool employingEVs derived from MSCs to deliver therapeuticfactors for liver fibrosis therapy. First, we compared ascalable method by ion exchange chromatography toisolate EVs from HUCPVC conditioned media with classicultracentrifugation method. By the chromatograph,EVs was collected in 3 elution fractions, showed typicaland homogeneous morphology, and CD9, CD63 andCD81 markers expression. Second, we evaluated thetherapeutic potential on liver fibrosis of EVs derived fromdifferent clinically relevant sources of MSC; adipose tissue(ASC-EVs), HUCPVC-EVs and induced pluripotentstem cells-derived MSC (iMSC-EV). In vitro EVs antifibroticeffect was tested on hepatic stellate cells (HSC)line. After incubation, EVs from different MSC sourcesdown-regulated pro-fibrogenic genes Col1a2 and α-SMAexpression in similar levels. Then, the therapeutic potentialof EVs was compared on experimental mice model ofliver fibrosis (thioacetamide for 8 weeks in BALB/c mice).On week 6, ASC-EV, iMSC-EV and HUCPVC-EV werei.v. injected every 5 days for a total of 3 doses. At week8, animals were sacrificed, and liver samples analyzed.The treatment with the three different EVs decreased collagendeposit and αSMA levels in liver tissue. In addition,EVs from different sources induce the hepatocellularproliferation compared with vehicle. Third, we confirmedby ELISA that chromatograph isolated’ EVs derived fromAdIGF-I-HUCPVC transport IGF1. Incubation of HSCswith EV-IGF1 resulted in downregulation of Col1a2 andαSMA expression demonstrating a reduction of its activationstatus. Finally, in vivo treatment with EV-IGF1 resultedin a further amelioration of collagen deposition andαSMA levels in liver tissue in comparison with controls.Consistently, an increase of PCNA+ cells after EV-IGF1application show the induction of liver regeneration. Conclusion:EVs derived from HUCPVCs load and transporttherapeutic factors, enhancing its anti-fibrotic and pro-regenerativepotential. The scalable chromatographicmethod retained the therapeutics potential of engineeredEV, emerging as an alternative for the treatment of liverdisease.