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

Cells

Mateusz Jeż¹  Olga Mucha¹  Józef Dulak¹  Krzysztof Szade¹  Kalina Andrysiak¹  Alicja Martyniak¹  Jacek Stępniewski¹  AnnaM. Sanetra²  Alan Kania²  Katarzyna Palus-Chramiec²  MarianH. Lewandowski²  Łukasz Chrobok²  Ewelina Pośpiech³ 

[1] Department of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Krakow, Poland;Department of Neurophysiology and Chronobiology, Institute of Zoology and Biomedical Research, Jagiellonian University, 30-387 Krakow, Poland;Human Genome Variation Research Group, Malopolska Centre of Biotechnology, Jagiellonian University, 30-387 Krakow, Poland;

关键词: hiPSC-CMs;    HO-1;    CRISPR/Cas9;    electrophysiology;    hypertrophy;    IGF2;   

DOI  :  10.3390/cells10030522

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown