Cardiac protection induced by urocortin-2 enables the regulation of apoptosis and fibrosis after ischemia and reperfusion involving miR-29a modulation

Isabel Mayoral-González; Eva M Calderón-Sánchez; Isabel Galeano-Otero; Marta Martín-Bórnez; Encarnación Gutiérrez-Carretero; María Fernández-Velasco; Nieves Domenech; María Generosa Crespo-Leiro; Ana María Gómez; Antonio Ordóñez-Fernández; Abdelkrim Hmadcha; Tarik Smani

doi:10.1016/j.omtn.2022.01.003

Cardiac protection induced by urocortin-2 enables the regulation of apoptosis and fibrosis after ischemia and reperfusion involving miR-29a modulation

Mol Ther Nucleic Acids. 2022 Jan 10:27:838-853. doi: 10.1016/j.omtn.2022.01.003. eCollection 2022 Mar 8.

Authors

Isabel Mayoral-González^{1

2}, Eva M Calderón-Sánchez^{2

3}, Isabel Galeano-Otero^{2

4}, Marta Martín-Bórnez^{1

2}, Encarnación Gutiérrez-Carretero^{1

2

3}, María Fernández-Velasco^{3

5}, Nieves Domenech^{3

6}, María Generosa Crespo-Leiro^{3

6}, Ana María Gómez⁷, Antonio Ordóñez-Fernández^{1

2

3}, Abdelkrim Hmadcha^{8

9}, Tarik Smani^{2

3

4}

Affiliations

¹ Department of Surgery, University of Seville, Seville, Spain.
² Group of Cardiovascular Pathophysiology, Institute of Biomedicine of Seville (IBiS), University Hospital of Virgen del Rocío/University of Seville/CSIC, Seville, Spain.
³ Centro de Investigación Biomédica en Red Enfermedades Cardiovaculares (CIBERCV), Madrid, Spain.
⁴ Department of Medical Physiology and Biophysics, University of Seville, Seville, Spain.
⁵ Innate Immune Response Group, IdiPAZ, La Paz University Hospital, Madrid, Spain.
⁶ Cardiology Department, Instituto de Investigación Biomédica de A Coruña, Complexo Hospitalario Universitario de A Coruña, Servicio Gallego de Salud, Universidade da Coruña, Coruña, Spain.
⁷ Signaling and Cardiovascular Pathophysiology, INSERM, Université Paris Saclay, Châtenay-Malabry, France.
⁸ Department of Biotechnology, University of Alicante, Alicante, Spain.
⁹ University of Pablo Olavide, Seville, Spain.

Abstract

Urocortin-2 (Ucn-2) has demonstrated cardioprotective actions against myocardial ischemia-reperfusion (I/R) injuries. Herein, we explored the protective role of Ucn-2 through microRNAs (miRNAs) post-transcriptional regulation of apoptotic and pro-fibrotic genes. We determined that the intravenous administration of Ucn-2 before heart reperfusion in a Wistar rat model of I/R recovered cardiac contractility and decreased fibrosis, lactate dehydrogenase release, and apoptosis. The infusion of Ucn-2 also inhibited the upregulation of 6 miRNAs in revascularized heart. The in silico analysis indicated that miR-29a and miR-451_1∗ are predicted to target many apoptotic and fibrotic genes. Accordingly, the transfection of neonatal rat ventricular myocytes with mimics overexpressing miR-29a, but not miR-451_1∗, prevented I/R-induced expression of pro- and anti-apoptotic genes such as Apaf-1, Hmox-1, and Cycs, as well as pro-fibrotic genes Col-I and Col-III. We also confirmed that Hmox-1, target of miR-29a, is highly expressed at the mRNA and protein levels in adult rat heart under I/R, whereas, Ucn-2 abolished I/R-induced mRNA and protein upregulation of HMOX-1. Interestingly, a significant upregulation of Hmox-1 was observed in the ventricle of ischemic patients with heart failure, correlating negatively with the left ventricle ejection fraction. Altogether, these data indicate that Ucn-2, through miR-29a regulation, provides long-lasting cardioprotection, involving the post-transcriptional regulation of apoptotic and fibrotic genes.

Keywords: Ucn-2; apoptosis; cardiac remodeling; fibrosis; heart failure; ischemia and reperfusion; miRNA.