Use of FDSS/μCell imaging platform for preclinical cardiac electrophysiology safety screening of compounds in human induced pluripotent stem cell-derived cardiomyocytes
Use of FDSS/μCell imaging platform for preclinical cardiac electrophysiology safety screening of compounds in human induced pluripotent stem cell-derived cardiomyocytes
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DOI：
10.1016/j.vascn.2016.05.009
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发表时间：
2016-09-01
影响因子：
1.9
通讯作者：
Sannajust, Frederick
中科院分区：
文献类型：
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作者：
Zeng, Haoyu;Roman, Maria I.;Sannajust, Frederick
FDSS/mu Cell是一种高速采集成像平台（Hamamatsu Ltd.，Hamamatsu，日本），其允许在受控条件下同时进行高通量阅读。我们评估了在存在或不存在已知干扰心脏离子通道的44种药理学试剂（例如，hERG、IKs、NaV1.5、CaV1.2）。我们测试了两种Ca 2+敏感的荧光染料（Codex ACTOne（R）和EarlyTox（R））和一种膜电位染料（FLIPR（R）膜电位染料）。我们能够定量和报告药物诱导的早期后除极（EAD）样波形、心肌细胞异位搏动和急性作用（1小时内）的药物亚组的搏动率变化。心血管药物如多非利特和D，L-索他洛尔分别在3 nM和10 μ M时显示EAD样信号。中枢神经系统药物，如氟哌啶醇和舍吲哚，表现出EAD样信号和异位搏动在30 nM和1 μ M，分别。其他药物，如阿司咪唑，索利那新，和阿托沙星，分别在30 nM，3 μ M和300 μ M时表现出类似的心律失常。我们的数据表明，膜电位和细胞内Ca 2+信号是紧密耦合的，这支持了以下观点：报告的EAD样信号是心脏动作电位EAD信号的准确代表。最后，EAD样Ca 2+信号与临床相关浓度密切相关，在健康志愿者口服我们测试的某些化合物时观察到尖端扭转型室性心动过速（TdPs），如PharmaPendium（R）所报道。(C)2016 Elsevier Inc. All rights reserved.
FDSS/mu Cell is a high-speed acquisition imaging platform (Hamamatsu Ltd., Hamamatsu, Japan) that allows for simultaneous high-throughput reading under controlled conditions. We evaluated the Ca2+ transients or optical membrane potential changes of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) (iCells) in the presence or absence of 44 pharmacological agents known to interfere with cardiac ion channels (e.g., hERG, IKs, NaV1.5, CaV1.2). We tested two Ca2+-sensitive fluorescence dyes (Codex ACTOne (R) and EarlyTox (R)) and a membrane potential dye (FLIPR (R) membrane potential dye). Wewere able to quantify and report drug-induced early-after depolarizations (EAD)-like waveforms, cardiomyocyte ectopic beats and changes in beating rate froma subgroup of pharmacological agents acting acutely (within a 1-hour period). Cardiovascular drugs, such as dofetilide and D,L-sotalol, exhibited EAD-like signals at 3nMand 10 mu M, respectively. CNS drugs, such as haloperidol and sertindole, exhibited EAD-like signals and ectopic beats at 30 nMand 1 mu M, respectively. Other drugs, such as astemizole, solifenacin, andmoxifloxacin, exhibited similar arrhythmias at 30 nM, 3 mu Mand 300 mu M, respectively. Our data suggest that the membrane potential and intracellular Ca2+ signal are tightly coupled, supporting the idea that the EAD-like signals reported are the accurate representation of an EAD signal of the cardiac action potential. Finally, the EAD-like Ca2+ signal waswell correlated to clinically-relevant concentrationswhere Torsade de Pointes (TdPs) arrhythmiaswere noted in healthy volunteers treated orallywith some of the compounds we tested, as reported in PharmaPendium (R). (C) 2016 Elsevier Inc. All rights reserved.