Arrhythmia Assessment in Heterotypic Human Cardiac Myocyte-Fibroblast Microtissues.
Arrhythmia Assessment in Heterotypic Human Cardiac Myocyte-Fibroblast Microtissues.
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DOI：
10.1007/978-1-0716-2261-2_10
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发表时间：
2022
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化学诱导心律失常的风险评估试验至关重要，但目前的心脏毒性试验存在重大局限性，包括侧重于单一选择离子通道，体外和体内使用非人类物种，以及有限的直接生理转化。为了预测实际的不良临床药理学风险，化学品和药物的心律失常评估模型应适合目的，并适合于评估作用机制可能不完全已知的化合物。在这里，我们描述了使用纯化的人诱导多能干细胞（hiPSC）衍生的心肌细胞和人心脏成纤维细胞的3D人类心脏微组织模型有效和可靠地筛选致瘤性心脏毒性的方法。将电压和钙敏感染料的光学映射-一种评估心脏动作电位和钙瞬变的既定方法-应用于3D异型心肌细胞-成纤维细胞组织，可以生成大量单个微组织并对其进行功能分析，以提供更大的通量和高的分析统计功效。标准细胞培养板中的数百个微组织可以以低变异性逐拍、微组织间和跨hiPSC-心肌细胞分化批次产生，从而减少了每个条件下预测输出所需的微组织数量。本文所述的平台可用作敏感、有效和预测性的临床前模型，经验证可用于评估人类促流产风险。
Risk assessment assays for chemically induced arrhythmia are critical, but significant limitations exist with current cardiotoxicity testing, including a focus on single select ion channels, the use of non-human species in vitro and in vivo, and limited direct physiological translation. To be predictive of actual adverse clinical arrhythmic risk, arrhythmia assessment models for chemicals and drugs should be fit-for-purpose and suited for evaluating compounds in which the mechanism of action may not be entirely known. Here, we describe methods for efficient and reliable screening for arrhythmogenic cardiotoxicity with a 3D human cardiac microtissue model using purified human-induced pluripotent stem cell (hiPSC)-derived cardiomyocytes and human cardiac fibroblasts. Applying optical mapping of voltage and calcium-sensitive dyes—an established approach to evaluate cardiac action potentials and calcium transients—to 3D heterotypic cardiac myocyte–fibroblast tissues allows for the generation and functional analysis of a large number of individual microtissues to provide greater throughput and high statistical power in analyses. Hundreds of microtissues in standard cell culture plates can be produced with low variability beat-to-beat, microtissue-to-microtissue, and across hiPSC-cardiomyocyte differentiation batches, reducing the number of microtissues required per condition for predictive outputs. The platform described here can be used as a sensitive, efficient, and predictive preclinical model validated for the purpose of assessing human pro-arrhythmic risk.