Jans Danny, Callewaert Geert, Krylychkina Olga, Hoffman Luis, Gullo Francesco, Prodanov Dimiter, Braeken Dries
IMEC Life Science Technologies, Kapeldreef 75, B3001 Leuven, Belgium.
IMEC Life Science Technologies, Kapeldreef 75, B3001 Leuven, Belgium.
J Pharmacol Toxicol Methods. 2017 Sep;87:48-52. doi: 10.1016/j.vascn.2017.05.003. Epub 2017 May 23.
Drug-induced cardiotoxicity poses a negative impact on public health and drug development. Cardiac safety pharmacology issues urged for the preclinical assessment of drug-induced ventricular arrhythmia leading to the design of several in vitro electrophysiological screening assays. In general, patch clamp systems allow for intracellular recordings, while multi-electrode array (MEA) technology detect extracellular activity. Here, we demonstrate a complementary metal oxide semiconductor (CMOS)-based MEA system as a reliable platform for non-invasive, long-term intracellular recording of cardiac action potentials at high resolution. Quinidine (8 concentrations from 10 to 2.10M) and verapamil (7 concentrations from 10 to 10M) were tested for dose-dependent responses in a network of cardiomyocytes. Electrophysiological parameters, such as the action potential duration (APD), rates of depolarization and repolarization and beating frequency were assessed. In hiPSC, quinidine prolonged APD with EC of 2.2·10M. Further analysis indicated a multifactorial action potential prolongation by quinidine: (1) decreasing fast repolarization with IC of 1.1·10M; (2) reducing maximum upstroke velocity with IC of 2.6·10M; and (3) suppressing spontaneous activity with EC of 3.8·10M. In rat neonatal cardiomyocytes, verapamil blocked spontaneous activity with EC of 5.3·10M and prolonged the APD with EC of 2.5·10M. Verapamil reduced rates of fast depolarization and repolarization with ICs of 1.8 and 2.2·10M, respectively. In conclusion, the proposed action potential-based MEA platform offers high quality and stable long-term recordings with high information content allowing to characterize multi-ion channel blocking drugs. We anticipate application of the system as a screening platform to efficiently and cost-effectively test drugs for cardiac safety.
药物诱导的心脏毒性对公众健康和药物研发产生负面影响。心脏安全药理学问题促使人们对药物诱导的室性心律失常进行临床前评估，从而设计出多种体外电生理筛选试验。一般来说，膜片钳系统可进行细胞内记录，而多电极阵列（MEA）技术则检测细胞外活动。在此，我们展示了一种基于互补金属氧化物半导体（CMOS）的MEA系统，它是一个可靠的平台，可用于高分辨率、无创、长期的心脏动作电位细胞内记录。在心肌细胞网络中测试了奎尼丁（8种浓度，范围为10至2.10M）和维拉帕米（7种浓度，范围为10至10M）的剂量依赖性反应。评估了电生理参数，如动作电位持续时间（APD）、去极化和复极化速率以及搏动频率。在人诱导多能干细胞（hiPSC）中，奎尼丁使APD延长，其半数有效浓度（EC）为2.2·10M。进一步分析表明，奎尼丁对动作电位的延长具有多因素作用：（1）以1.1·10M的半数抑制浓度（IC）降低快速复极化；（2）以2.6·10M的IC降低最大上升速度；（3）以3.8·10M的EC抑制自发活动。在大鼠新生心肌细胞中，维拉帕米以5.3·10M的EC阻断自发活动，并以2.5·10M的EC延长APD。维拉帕米分别以1.8和2.2·10M的IC降低快速去极化和复极化速率。总之，所提出的基于动作电位的MEA平台提供高质量、稳定的长期记录，具有高信息含量，能够对多离子通道阻断药物进行表征。我们预期该系统可作为筛选平台，高效且经济地测试药物的心脏安全性。
