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Abstract：
Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) have emerged as a promising tool for studying cardiac physiology and drug responses. However, their use is largely limited by an immature phenotype and lack of high-throughput analytical methodology. In this study, we developed a high-throughput testing platform utilizing hPSC-CMs to assess the cardiotoxicity and effectiveness of drugs. Following an optimized differentiation and maturation protocol, hPSC-CMs exhibited mature CM morphology, phenotype, and functionality, making them suitable for drug testing applications. We monitored intracellular calcium dynamics using calcium imaging techniques to measure spontaneous calcium oscillations in hPSC-CMs in the presence or absence of test compounds. For the cardiotoxicity test, hPSC-CMs were treated with various compounds, and calcium flux was measured to evaluate their effects on calcium dynamics. We found that cardiotoxic drugs withdrawn due to adverse drug reactions, including encainide, mibefradil, and cetirizine, exhibited toxicity in hPSC-CMs but not in HEK293-hERG cells. Additionally, in the effectiveness test, hPSC-CMs were exposed to ATX-II, a sodium current inducer for mimicking long QT syndrome type 3, followed by exposure to test compounds. The observed changes in calcium dynamics following drug exposure demonstrated the utility of hPSC-CMs as a versatile model system for assessing both cardiotoxicity and drug efficacy. Overall, our findings highlight the potential of hPSC-CMs in advancing drug discovery and development, which offer a physiologically relevant platform for the preclinical screening of novel therapeutics.
摘要：
人类多能干细胞衍生的心肌细胞（hPSC-CM）已成为研究心脏生理学和药物反应的有希望的工具。然而,它们的使用在很大程度上受到不成熟表型和缺乏高通量分析方法的限制.在这项研究中,我们开发了一个利用hPSC-CM评估药物心脏毒性和有效性的高通量检测平台.经过优化的分化和成熟方案，hPSC-CM表现出成熟的CM形态，表型,和功能，使它们适用于药物测试应用。我们使用钙成像技术监测细胞内钙动力学，以在存在或不存在测试化合物的情况下测量hPSC-CM中的自发钙振荡。对于心脏毒性测试，用各种化合物处理hPSC-CM，并测量钙通量以评估其对钙动力学的影响。我们发现心脏毒性药物由于药物不良反应而停用，包括encainide,Mibefradil,还有西替利嗪,在hPSC-CM中表现出毒性，但在HEK293-hERG细胞中没有。此外,在有效性测试中，hPSC-CM暴露于ATX-II，钠电流诱导剂，用于模拟3型长QT综合征，然后暴露于测试化合物。观察到的药物暴露后钙动力学的变化证明了hPSC-CM作为评估心脏毒性和药物功效的通用模型系统的实用性。总的来说,我们的发现强调了hPSC-CM在推进药物发现和开发方面的潜力，这为新疗法的临床前筛选提供了生理相关的平台。
