Role of Heme-Oxygenase-1 in Biology of Cardiomyocytes Derived from Human Induced Pluripotent Stem Cells.

Mateusz Jeż,Krzysztof Szade,Kalina Andrysiak,Ewelina Pośpiech,Olga Mucha,Alicja Martyniak,Józef Dulak,Marian H Lewandowski,Anna M Sanetra,Katarzyna Palus-Chramiec,Jacek Stępniewski,Łukasz Chrobok,Alan Kania

doi:10.3390/cells10030522

Abstract

Heme oxygenase-1 (HO-1, encoded by HMOX1) is a cytoprotective enzyme degrading heme into CO, Fe2+, and biliverdin. HO-1 was demonstrated to affect cardiac differentiation of murine pluripotent stem cells (PSCs), regulate the metabolism of murine adult cardiomyocytes, and influence regeneration of infarcted myocardium in mice. However, the enzyme’s effect on human cardiogenesis and human cardiomyocytes’ electromechanical properties has not been described so far. Thus, this study aimed to investigate the role of HO-1 in the differentiation of human induced pluripotent stem cells (hiPSCs) into hiPSC-derived cardiomyocytes (hiPSC-CMs). hiPSCs were generated from human fibroblasts and peripheral blood mononuclear cells using Sendai vectors and subjected to CRISPR/Cas9-mediated HMOX1 knock-out. After confirming lack of HO-1 expression on the protein level, isogenic control and HO-1-deficient hiPSCs were differentiated into hiPSC-CMs. No differences in differentiation efficiency and hiPSC-CMs metabolism were observed in both cell types. The global transcriptomic analysis revealed, on the other hand, alterations in electrophysiological pathways in hiPSC-CMs devoid of HO-1, which also demonstrated increased size. Functional consequences in changes in expression of ion channels genes were then confirmed by patch-clamp analysis. To the best of our knowledge, this is the first report demonstrating the link between HO-1 and electrophysiology in human cardiomyocytes.

Highlights

The leading cause of death in developed countries, despite significant advances in pharmacotherapy and cardiac surgery, are cardiovascular diseases, including ischemic heart diseases (IHD) [1]
Human Induced Pluripotent Stem Cells Initiative’s collection. human induced pluripotent stem cells (hiPSCs).1 was reprogrammed from commercially available BJ fibroblasts (ATCC, CRL-2522), and hiPSC.3 were reprogrammed from PBMCs isolated from a healthy donor
heme oxygenase-1 (HO-1) KO hiPSC clones were generated from the characterized lines through the nucleofection with a plasmid encoding Cas9 and designed sgRNAs targeting HMOX1 exon

Summary

Introduction

The leading cause of death in developed countries, despite significant advances in pharmacotherapy and cardiac surgery, are cardiovascular diseases, including ischemic heart diseases (IHD) [1]. Myocardial infarction (MI) leads to ischemia and hypoxia of the heart muscle, resulting in loss of up to one billion cardiomyocytes Due to the adult human heart’s negligible regenerative capabilities, a scar is formed in place of lost cardiomyocytes. A non-contractile collagen tissue further weakens electromechanical properties of a damaged hearts, which may lead to arrhythmias, severe failure, and eventually, death [3]. Despite significant advances in cardiology, organ transplantation is the only solution for many patients with chronic heart failure. A promising alternative to heart transplantations may be a rapidly growing field of personalized and regenerative medicine

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