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.
Cells. 2021 Mar 1;10(3):522. doi: 10.3390/cells10030522.
Heme oxygenase-1 (HO-1, encoded by ) is a cytoprotective enzyme degrading heme into CO, Fe, 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 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.
血红素加氧酶-1（HO-1，由 编码）是一种细胞保护性酶，可将血红素降解为 CO、Fe 和胆红素。HO-1 被证明可影响鼠多能干细胞（PSCs）的心脏分化，调节鼠成年心肌细胞的代谢，并影响小鼠梗死心肌的再生。然而，迄今为止，该酶对人类心脏发生和人心肌细胞机电特性的影响尚未描述。因此，本研究旨在探讨 HO-1 在人诱导多能干细胞（hiPSCs）分化为 hiPSC 衍生的心肌细胞（hiPSC-CMs）中的作用。使用 Sendai 载体从人成纤维细胞和外周血单核细胞中生成 hiPSCs，并通过 CRISPR/Cas9 介导的 敲除。在确认蛋白质水平上缺乏 HO-1 表达后，对同基因对照和 HO-1 缺陷型 hiPSCs 进行分化为 hiPSC-CMs。在这两种细胞类型中，分化效率和 hiPSC-CMs 代谢没有差异。另一方面，全转录组分析显示，缺乏 HO-1 的 hiPSC-CMs 中存在电生理途径的改变，这也表明其大小增加。通过膜片钳分析进一步证实了离子通道基因表达变化的功能后果。据我们所知，这是第一篇报道 HO-1 与人心肌细胞电生理学之间存在联系的文章。
