GMP mesenchymal stromal cell-derived extracellular vesicles cells for cardiac tissue repair after myocardial infarction

• Grant number: 21/13027-0
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• Start date: August 01, 2022
• End date: July 31, 2023
• Field of knowledge: Health Sciences - Medicine - Medical Clinics
• Principal Investigator: Katashi Okoshi
• Grantee: Luana Urbano Pagan
• Supervisor: Carolina Galvez-Monton
• Host Institution: Faculdade de Medicina (FMB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
• Institution abroad: Instituto de Investigación Germans Trias i Pujol (IGTP), Spain
• Associated to the scholarship: 18/07048-2 - Influence of empagliflozin on the myocardium of rats with heart failure induced by aortic stenosis, BP.PD

Abstract

Myocardial infarction (MI) is among the main cardiovascular diseases. After MI, part of the myocardial tissue is not efficient for regeneration into new functional cardiac tissue. Instead, a scar is formed in the infarcted area. The extension of the formed scar determines the tendency to adverse cardiac remodeling. Given the limited capacity for self-renewal, cell-based strategies to regenerate lost cardiomyocytes or to promote endogenous repair became important, and the interesse in tissue engineering therapies for the treatment of MI has increased. In this context, mesenchymal stromal cells (MSC) cultured in vitro, mainly from cardiac adipose tissue biopsies, have been studied. Extracellular vesicles (EV) are active factors secreted by MSCs, which play a role in paracrine signaling. However, their mechanism of action, the activated signaling pathways, and the optimal administration route to allow the EV dose to be locally sufficient for the beneficial effects remain unknown. In this sense, this research project aims to isolate the EV of human Wharton's jelly-derived MSC (MSC-EV) under Good Manufacturing Practices (GMP) and evaluate their anti-inflammatory effects in vitro. Furthermore, produce cardiac scaffolds with MSC-EV, implant them in a porcine model of MI, and assess cardiac function and mortality. Methods: Production and isolation of extracellular vesicles from WJ MSC will be performed under GMP conditions. Transcriptomic and proteomic analysis will be performed to distinguish functional molecules contained within EV. The levels of circulating cytokines will be measured in the supernatants of cell culture by ELISA. Decellularized pericardial scaffolds will be obtained from the BST's Tissue Bank. Scaffold replenishment will be carried out seeding EV from 20 or 40 million producing MSC, WJ mixed in a GMP-grade peptide hydrogel. The pericardium graft enriched with MSC, WJ-EV will be implanted in a porcine MI model. Forty animals will be divided in four groups: MI + no scaffold implantation (n=10), MI + scaffold implantation (n=10), MI + scaffold with low EV dose (n=10) and MI + scaffold high EV dose (n=10). Cardiac function will be assessed by magnetic resonance imaging. Animals will be euthanized 1 month after MI induction. Tissues will be collected for analysis in future studies. Overall data from all planned objectives will be analysed statistically with the corresponding most suitable statistical analysis according to data type and normality distribution. Significance level will be set at 5%. (AU)