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对来自2家主要供应商的诱导多能干细胞衍生的人类心肌细胞批次差异的评估。

Evaluation of Batch Variations in Induced Pluripotent Stem Cell-Derived Human Cardiomyocytes from 2 Major Suppliers.

作者信息

Huo Jianhua, Kamalakar Archana, Yang Xi, Word Beverly, Stockbridge Norman, Lyn-Cook Beverly, Pang Li

机构信息

Division of Biochemical Toxicology, National Center for Toxicological Research, U.S. FDA, Jefferson, Arkansas 72079.

Department of Cardiovascular Medicine, First Hospital of Xi'an Jiaotong University, 277 Yanta West Road, Xi'an, ShaanXI, 710061, China.

出版信息

Toxicol Sci. 2017 Mar 1;156(1):25-38. doi: 10.1093/toxsci/kfw235.

Abstract

Drug-induced proarrhythmia is a major safety issue in drug development. Developing sensitive in vitro assays that can predict drug-induced cardiotoxicity in humans has been a challenge of toxicology research for decades. Recently, induced pluripotent stem cell-derived human cardiomyocytes (iPSC-hCMs) have become a promising model because they largely replicate the electrophysiological behavior of human ventricular cardiomyocytes. Patient-specific iPSC-hCMs have been proposed for personalized cardiac drug selection and adverse drug response prediction; however, many procedures are involved in cardiomyocytes differentiation and purification process, which may result in large line-to-line and batch-to-batch variations. Here, we examined the purity, cardiac ion channel gene expression profile, and electrophysiological response of 3 batches of iPSC-hCMs from each of 2 major cell suppliers. We found that iPSC-hCMs from both vendors had similar purities. Most of the cardiac ion channel genes were expressed uniformly among different batches of iCells, while larger variations were found in Cor.4U cells, particularly in the expression of CACNA1C, KCND2, and KCNA5 genes, which could underlie the differences in baseline beating rate (BR) and field potential duration (FPD) measurements. Although, in general, the electrophysiological responses of different batches of cells to Na+, Ca2+, Ikr, and Iks channel blockers were similar, with Ikr blocker-induced proarrhythmia, the sensitivities were depended on baseline BR and FPD values: cells that beat slower had longer FPD and greater sensitivity to drug-induced proarrhythmia. Careful evaluation of the performance of iPSC-hCMs and methods of data analysis is warranted for shaping regulatory standards in qualifying iPSC-hCMs for drug safety testing.

摘要

药物诱导的心律失常是药物研发中的一个主要安全问题。几十年来,开发能够预测人类药物诱导心脏毒性的灵敏体外试验一直是毒理学研究的一项挑战。最近,诱导多能干细胞衍生的人类心肌细胞(iPSC-hCMs)已成为一种有前景的模型,因为它们在很大程度上复制了人类心室心肌细胞的电生理行为。已有人提出使用患者特异性iPSC-hCMs进行个性化心脏药物选择和药物不良反应预测;然而,心肌细胞分化和纯化过程涉及许多步骤,这可能导致不同细胞系和批次之间存在较大差异。在此,我们检测了来自2家主要细胞供应商的3批次iPSC-hCMs的纯度、心脏离子通道基因表达谱和电生理反应。我们发现,两家供应商的iPSC-hCMs纯度相似。大多数心脏离子通道基因在不同批次的i细胞中表达一致,而在Cor.4U细胞中发现了较大差异,特别是在CACNA1C、KCND2和KCNA5基因的表达上,这可能是基线搏动率(BR)和场电位持续时间(FPD)测量结果存在差异的原因。尽管一般来说,不同批次的细胞对Na+、Ca2+、Ikr和Iks通道阻滞剂的电生理反应相似,但对于Ikr阻滞剂诱导的心律失常,其敏感性取决于基线BR和FPD值:搏动较慢的细胞FPD较长,对药物诱导的心律失常更敏感。为了制定iPSC-hCMs用于药物安全性测试的监管标准,有必要仔细评估iPSC-hCMs的性能和数据分析方法。

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