The action potential and comparative pharmacology of stem cell-derived human cardiomyocytes
The action potential and comparative pharmacology of stem cell-derived human cardiomyocytes
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DOI：
10.1016/j.vascn.2010.01.014
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发表时间：
2010-05-01
影响因子：
1.9
通讯作者：
Bruening-Wright, Andrew
中科院分区：
文献类型：
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作者：
Peng, Shengde;Lacerda, Antonio E.;Bruening-Wright, Andrew
干细胞来源的人心肌细胞（SC-hCMs）的心脏动作电位（CAP）可能是人类心脏安全性和有效性的最强大的临床前生物标志物。我们的实验通过检查这些细胞的CAP和相关药理学来验证这一假设。方法：将SC-hCMs与家兔和犬浦肯野纤维（PFs）的电生理和药理特性进行比较。心室SC-hCMs在心室、心房和结型心肌细胞（CMs）群体中提供了主要的电生理表型（类似于82%）。在SC-hCMs中，采用穿孔贴片、电流钳记录测量参比化合物的作用。选择I(Kr)， I(Ks)， I(Ca，L)， I（Na）和去甲肾上腺素（NE）抑制剂对SC-hCM动作电位（APs）进行检测。结果：暴露于hERG通道阻滞剂（特非那定、奎尼丁、西沙比利、索他洛尔、E-4031和维拉帕米）后，观察到AP延长，潜伏期明显短于PF试验。对于致反性化合物，特非那定和奎尼丁，SC-hCM AP延长的浓度明显低于犬或兔的PF AP。此外，I（Ks）阻滞剂chromanol 2936延长了sc - hcm的APs，而在缺乏肾上腺素能预处理的情况下，兔和犬的PF试验对I（Ks）阻滞剂不敏感。在SC-hCM实验中，100 nM E-4031和100 nM西沙匹利诱导了早期后去极化（EADs），但在犬和兔的PF实验中没有。选择性抑制I（Na）和I（Ca，L）分别减缓V（max）和缩短AP持续时间。NE延长sc - hcm的AP持续时间。讨论：SC-hCMs的CAP已被验证为心脏安全性和有效性的强大临床前生物标志物。除了它的人性，SC-hCM AP分析消除了扩散延迟，减少了测试化合物的消耗，证明了比传统的兔或犬PF分析更大的总体药理学敏感性，并准确地预测了已知的变性化合物的心脏风险。(C) 2010爱思唯尔公司版权所有。
Introduction: The cardiac action potential (CAP) of stem cell-derived human cardiomyocytes (SC-hCMs) is potentially the most powerful preclinical biomarker for cardiac safety and efficacy in humans. Our experiments tested this hypothesis by examining the CAP and relevant pharmacology of these cells. Methods: The electrophysiological and pharmacological profiles of SC-hCMs were compared to rabbit and canine Purkinje fibers (PFs). Ventricular SC-hCMs provided the dominant electrophysiological phenotype (similar to 82%) in a population of ventricular, atrial and nodal cardiomyocytes (CMs). The effects of reference compounds were measured in SC-hCMs using perforated patch, current clamp recording. Selective inhibitors of I(Kr), I(Ks), I(Ca,L), and I(Na), and norepinephrine (NE), were tested on SC-hCM action potentials (APs). Results: AP prolongation was observed upon exposure to hERG channel blockers (terfenadine, quinidine, cisapride, sotalol, E-4031 and verapamil), with significantly shorter latencies than in PF assays. For the torsadogenic compounds, terfenadine and quinidine, SC-hCM AP prolongation occurred at significantly lower concentrations than in canine or rabbit PF APs. Moreover, the I(Ks) blocker chromanol 2936 prolonged APs from SC-hCMs, whereas both rabbit and canine PF assays are insensitive to I(Ks) blockers in the absence of adrenergic preconditioning. Early afterdepolarizations (EADs) were induced by 100 nM E-4031 and 100 nM cisapride in the SC-hCM assay, but not in the canine or rabbit PF assay. Selective inhibition of I(Na) and I(Ca,L) slowed V(max) and shortened AP duration, respectively. NE prolonged the AP duration of SC-hCMs. Discussion: The CAP of SC-hCMs has been validated as a powerful preclinical biomarker for cardiac safety and efficacy. In addition to its human nature, the SC-hCM AP assay removes diffusion delays, reduces test compound consumption, demonstrates an overall pharmacological sensitivity that is greater than conventional rabbit or canine PF assays, and accurately predicts cardiac risk of known torsadogenic compounds. (C) 2010 Elsevier Inc. All rights reserved.