人诱导多能干细胞来源的心房样和心室样心肌细胞中的 Na1.5 通道的生物物理特性。
Biophysical properties of Na1.5 channels from atrial-like and ventricular-like cardiomyocytes derived from human induced pluripotent stem cells.
机构信息
Department of Medicine, Laval University, Quebec City, QC, Canada.
CERVO Brain Research Centre, 2601, chemin de la Canardière, Quebec City, QC, G1J 2G3, Canada.
出版信息
Sci Rep. 2023 Nov 24;13(1):20685. doi: 10.1038/s41598-023-47310-6.
Generating atrial-like cardiomyocytes derived from human induced pluripotent stem cells (hiPSCs) is crucial for modeling and treating atrial-related diseases, such as atrial arrythmias including atrial fibrillations. However, it is essential to obtain a comprehensive understanding of the electrophysiological properties of these cells. The objective of the present study was to investigate the molecular, electrical, and biophysical properties of several ion channels, especially Na1.5 channels, in atrial hiPSC cardiomyocytes. Atrial cardiomyocytes were obtained by the differentiation of hiPSCs treated with retinoic acid (RA). The quality of the atrial specification was assessed by qPCR, immunocytofluorescence, and western blotting. The electrophysiological properties of action potentials (APs), Ca dynamics, K and Na currents were investigated using patch-clamp and optical mapping approaches. We evaluated mRNA transcript and protein expressions to show that atrial cardiomyocytes expressed higher atrial- and sinoatrial-specific markers (MYL7, CACNA1D) and lower ventricular-specific markers (MYL2, CACNA1C, GJA1) than ventricular cardiomyocytes. The amplitude, duration, and steady-state phase of APs in atrial cardiomyocytes decreased, and had a shape similar to that of mature atrial cardiomyocytes. Interestingly, Na1.5 channels in atrial cardiomyocytes exhibited lower mRNA transcripts and protein expression, which could explain the lower current densities recorded by patch-clamp. Moreover, Na currents exhibited differences in activation and inactivation parameters. These differences could be explained by an increase in SCN2B regulatory subunit expression and a decrease in SCN1B and SCN4B regulatory subunit expressions. Our results show that a RA treatment made it possible to obtain atrial cardiomyocytes and investigate differences in Na1.5 channel properties between ventricular- and atrial-like cells.
从人诱导多能干细胞(hiPSCs)生成类心房肌细胞对于模拟和治疗与心房相关的疾病(如包括心房颤动在内的心房性心律失常)至关重要。然而,获得对这些细胞电生理特性的全面了解是至关重要的。本研究旨在研究几种离子通道,特别是 Na1.5 通道,在人心房 hiPSC 心肌细胞中的分子、电和生物物理特性。通过用视黄酸(RA)处理 hiPSCs 来获得心房肌细胞。通过 qPCR、免疫细胞荧光和 Western blot 评估心房特异性的质量。使用膜片钳和光学映射方法研究动作电位(APs)、Ca 动力学、K 和 Na 电流的电生理特性。我们评估了 mRNA 转录本和蛋白质表达,以表明心房肌细胞表达更高的心房和窦房结特异性标志物(MYL7、CACNA1D)和更低的心室特异性标志物(MYL2、CACNA1C、GJA1)比心室肌细胞。心房肌细胞中 AP 的幅度、持续时间和稳态相降低,并且形状类似于成熟的心房肌细胞。有趣的是,心房肌细胞中的 Na1.5 通道表现出较低的 mRNA 转录本和蛋白质表达,这可以解释通过膜片钳记录的较低电流密度。此外,Na 电流在激活和失活参数方面表现出差异。这些差异可以通过 SCN2B 调节亚基表达的增加和 SCN1B 和 SCN4B 调节亚基表达的减少来解释。我们的结果表明,RA 处理使得获得心房肌细胞并研究心室样细胞和心房样细胞之间 Na1.5 通道特性的差异成为可能。