Use of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes (hiPSC-CMs) to Monitor Compound Effects on Cardiac Myocyte Signaling Pathways.
Use of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes (hiPSC-CMs) to Monitor Compound Effects on Cardiac Myocyte Signaling Pathways.
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DOI：
10.1002/9780470559277.ch150035
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发表时间：
2015-09-01
影响因子：
--
通讯作者：
Davis M
中科院分区：
文献类型：
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作者：
Guo L;Eldridge S;Furniss M;Mussio J;Davis M
有必要开发基于机制的测定，以更好地告知心脏毒性的风险。人诱导多能干细胞衍生的心肌细胞（hiPSC-CM）作为用于药物发现和心脏毒性筛选的生物学相关体外模型正在获得快速接受。利用hiPSC-CM进行机制研究将受益于已知调节心肌细胞活力和功能的细胞途径的表达和活性的确认。本单元描述了一种方法来证明hiPSC-CM中信号通路的存在和功能以及治疗对这些通路的影响。我们提出了一个工作流程，该工作流程采用协议来证明蛋白质表达，感兴趣的信号传导途径的功能完整性，并表征信号调节的生物学后果。这些方案利用结构、功能和生化终点的独特组合来询问化合物对心肌细胞的作用。
There is a need to develop mechanism-based assays to better inform risk of cardiotoxicity. Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) are gaining rapid acceptance as a biologically relevant in vitro model for use in drug discovery and cardiotoxicity screens. Utilization of hiPSC-CMs for mechanistic investigations would benefit from confirmation of the expression and activity of cellular pathways that are known to regulate cardiac myocyte viability and function. This unit describes an approach to demonstrate the presence and function of signaling pathway(s) in hiPSC-CMs and the effects of treatments on these pathways. We present a workflow that employs protocols to demonstrate protein expression, functional integrity of signaling pathway(s) of interest and that characterize biological consequences of signaling modulation. These protocols utilize a unique combination of structural, functional and biochemical endpoints to interrogate compound effects on cardiomyocytes.