Combined genetic and chemical screens indicate protective potential for EGFR inhibition to cardiomyocytes under hypoxia

Klaus Elenius,Ilkka Paatero,Juho Heliste,Katri Vaparanta,Anne Jokilammi

doi:10.1038/s41598-021-96033-z

Abstract

The return of blood flow to ischemic heart after myocardial infarction causes ischemia–reperfusion injury. There is a clinical need for novel therapeutic targets to treat myocardial ischemia–reperfusion injury. Here we screened for targets for the treatment of ischemia–reperfusion injury using a combination of shRNA and drug library analyses in HL-1 mouse cardiomyocytes subjected to hypoxia and reoxygenation. The shRNA library included lentiviral constructs targeting 4625 genes and the drug library 689 chemical compounds approved by the Food and Drug Administration (FDA). Data were analyzed using protein–protein interaction and pathway analyses. EGFR inhibition was identified as a cardioprotective mechanism in both approaches. Inhibition of EGFR kinase activity with gefitinib improved cardiomyocyte viability in vitro. In addition, gefitinib preserved cardiac contractility in zebrafish embryos exposed to hypoxia-reoxygenation in vivo. These findings indicate that the EGFR inhibitor gefitinib is a potential candidate for further studies of repurposing the drug for the treatment of myocardial infarction.

Highlights

The return of blood flow to ischemic heart after myocardial infarction causes ischemia–reperfusion injury
The cells were subjected into a dropout screening approach, in which the enrichment or depletion of individual short hairpin RNAs (shRNA) species in the cells surviving the hypoxia-reoxygenation treatment was assessed by comparing to quantities of the same shRNAs in cells cultured in normoxia
We screened for novel therapeutic targets for myocardial ischemia–reperfusion injury using a combination of systematic functional screens based on shRNA and drug libraries in cardiomyocytes subjected to experimental hypoxia-reoxygenation

Summary

Introduction

The return of blood flow to ischemic heart after myocardial infarction causes ischemia–reperfusion injury. We screened for targets for the treatment of ischemia–reperfusion injury using a combination of shRNA and drug library analyses in HL-1 mouse cardiomyocytes subjected to hypoxia and reoxygenation. Gefitinib preserved cardiac contractility in zebrafish embryos exposed to hypoxia-reoxygenation in vivo These findings indicate that the EGFR inhibitor gefitinib is a potential candidate for further studies of repurposing the drug for the treatment of myocardial infarction. We used a combination of two functional screens, one based on an shRNA library targeting 4625 genes and the other on a chemical library including 689 FDA-approved drugs, to screen for targets of myocardial ischemia–reperfusion injury using HL-1 cardiomyocytes exposed to hypoxia and reoxygenation. Protein–protein interaction and pathway analyses of the top hits from both screens suggested EGFR inhibition as a potential mechanism to enhance the survival of hypoxic cardiomyocytes. EGFR inhibitors may represent a novel approach to be investigated for the treatment of myocardial ischemia–reperfusion injury

Methods

Results

Conclusion