Abstract：
BACKGROUND: Cardiac safety assessment, such as lethal arrhythmias and contractility dysfunction, is critical during drug development. Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) have been shown to be useful in predicting drug-induced proarrhythmic risk through international validation studies. Although cardiac contractility is another key function, fit-for-purpose hiPSC-CMs in evaluating drug-induced contractile dysfunction remain poorly understood. In this study, we investigated whether alignment of hiPSC-CMs on nanopatterned culture plates can assess drug-induced contractile changes more efficiently than non-aligned monolayer culture.
METHODS: Aligned hiPSC-CMs were obtained by culturing on 96-well culture plates with a ridge-groove-ridge nanopattern on the bottom surface, while non-aligned hiPSC-CMs were cultured on regular 96-well plates. Next-generation sequencing and qPCR experiments were performed for gene expression analysis. Contractility of the hiPSC-CMs was assessed using an imaging-based motion analysis system.
RESULTS: When cultured on nanopatterned plates, hiPSC-CMs exhibited an aligned morphology and enhanced expression of genes encoding proteins that regulate contractility, including myosin heavy chain, calcium channel, and ryanodine receptor. Compared to cultures on regular plates, the aligned hiPSC-CMs also showed both enhanced contraction and relaxation velocity. In addition, the aligned hiPSC-CMs showed a more physiological response to positive and negative inotropic agents, such as isoproterenol and verapamil.
CONCLUSIONS: Taken together, the aligned hiPSC-CMs exhibited enhanced structural and functional properties, leading to an improved capacity for contractility assessment compared to the non-aligned cells. These findings suggest that the aligned hiPSC-CMs can be used to evaluate drug-induced cardiac contractile changes.
METHODS: Aligned hiPSC-CMs were obtained by culturing on 96-well culture plates with a ridge-groove-ridge nanopattern on the bottom surface, while non-aligned hiPSC-CMs were cultured on regular 96-well plates. Next-generation sequencing and qPCR experiments were performed for gene expression analysis. Contractility of the hiPSC-CMs was assessed using an imaging-based motion analysis system.
RESULTS: When cultured on nanopatterned plates, hiPSC-CMs exhibited an aligned morphology and enhanced expression of genes encoding proteins that regulate contractility, including myosin heavy chain, calcium channel, and ryanodine receptor. Compared to cultures on regular plates, the aligned hiPSC-CMs also showed both enhanced contraction and relaxation velocity. In addition, the aligned hiPSC-CMs showed a more physiological response to positive and negative inotropic agents, such as isoproterenol and verapamil.
CONCLUSIONS: Taken together, the aligned hiPSC-CMs exhibited enhanced structural and functional properties, leading to an improved capacity for contractility assessment compared to the non-aligned cells. These findings suggest that the aligned hiPSC-CMs can be used to evaluate drug-induced cardiac contractile changes.
摘要：
背景：心脏安全性评估，如致命的心律失常和收缩功能障碍，在药物开发过程中至关重要。通过国际验证研究，人类诱导的多能干细胞衍生的心肌细胞（hiPSC-CM）已被证明可用于预测药物诱导的心律失常风险。虽然心脏收缩是另一个关键功能，适合用途的hiPSC-CM在评估药物诱导的收缩功能障碍方面仍然知之甚少。在这项研究中,我们调查了hiPSC-CM在纳米图案培养板上的排列是否比非排列单层培养更有效地评估药物诱导的收缩变化.
方法：通过在96孔培养板上培养底面上带有脊-凹槽-脊纳米图案，获得了对齐的hiPSC-CM，而非对齐的hiPSC-CM在常规96孔板上培养。进行下一代测序和qPCR实验用于基因表达分析。使用基于成像的运动分析系统评估hiPSC-CM的收缩性。
结果：在纳米图案板上培养时，hiPSC-CM表现出比对的形态和增强的基因表达编码调节收缩性的蛋白质，包括肌球蛋白重链,钙通道，还有ryanodine受体.与普通盘子上的培养物相比，对齐的hiPSC-CM也显示出增强的收缩和弛豫速度。此外,排列的hiPSC-CM对正性和负性肌力药物显示出更多的生理反应，如异丙肾上腺素和维拉帕米。
结论：综合来看，对齐的HiPSC-CM表现出增强的结构和功能特性，与非对齐细胞相比，提高了收缩力评估的能力。这些发现表明，对齐的hiPSC-CM可用于评估药物诱导的心脏收缩变化。
方法：通过在96孔培养板上培养底面上带有脊-凹槽-脊纳米图案，获得了对齐的hiPSC-CM，而非对齐的hiPSC-CM在常规96孔板上培养。进行下一代测序和qPCR实验用于基因表达分析。使用基于成像的运动分析系统评估hiPSC-CM的收缩性。
结果：在纳米图案板上培养时，hiPSC-CM表现出比对的形态和增强的基因表达编码调节收缩性的蛋白质，包括肌球蛋白重链,钙通道，还有ryanodine受体.与普通盘子上的培养物相比，对齐的hiPSC-CM也显示出增强的收缩和弛豫速度。此外,排列的hiPSC-CM对正性和负性肌力药物显示出更多的生理反应，如异丙肾上腺素和维拉帕米。
结论：综合来看，对齐的HiPSC-CM表现出增强的结构和功能特性，与非对齐细胞相比，提高了收缩力评估的能力。这些发现表明，对齐的hiPSC-CM可用于评估药物诱导的心脏收缩变化。
