Abstract：
Human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes (CMs) have found utility for conducting in vitro drug screening and disease modelling to gain crucial insights into pharmacology or disease phenotype. However, diseases such as atrial fibrillation, affecting >33 M people worldwide, demonstrate the need for cardiac subtype-specific cells. Here, we sought to investigate the base characteristics and pharmacological differences between commercially available chamber-specific atrial or ventricular hiPSC-CMs seeded onto ultra-thin, flexible PDMS membranes to simultaneously measure contractility in a 96 multi-well format. We investigated the effects of GPCR agonists (acetylcholine and carbachol), a Ca2+ channel agonist (S-Bay K8644), an HCN channel antagonist (ivabradine) and K+ channel antagonists (4-AP and vernakalant). We observed differential effects between atrial and ventricular hiPSC-CMs on contractile properties including beat rate, beat duration, contractile force and evidence of arrhythmias at a range of concentrations. As an excerpt of the compound analysis, S-Bay K8644 treatment showed an induced concentration-dependent transient increase in beat duration of atrial hiPSC-CMs, whereas ventricular cells showed a physiological increase in beat rate over time. Carbachol treatment produced marked effects on atrial cells, such as increased beat duration alongside a decrease in beat rate over time, but only minimal effects on ventricular cardiomyocytes. In the context of this chamber-specific pharmacology, we not only add to contractile characterization of hiPSC-CMs but propose a multi-well platform for medium-throughput early compound screening. Overall, these insights illustrate the key pharmacological differences between chamber-specific cardiomyocytes and their application on a multi-well contractility platform to gain insights for in vitro cardiac liability studies and disease modelling.
摘要：
人诱导多能干细胞（hiPSC）衍生的心肌细胞（CM）已发现可用于进行体外药物筛选和疾病建模，以获得对药理学或疾病表型的重要见解。然而,心房颤动等疾病,影响全球超过3300万人，证明需要心脏亚型特异性细胞。这里,我们试图调查市场上可买到的特定心房或心室hiPSC-CM的基本特征和药理学差异，柔性PDMS膜以96多孔格式同时测量收缩性。我们研究了GPCR激动剂（乙酰胆碱和卡巴胆碱）的作用，Ca2+通道激动剂（S-BayK8644），HCN通道拮抗剂（伊伐布雷定）和K通道拮抗剂（4-AP和vernakalant）。我们观察到心房和心室hiPSC-CM对收缩特性的不同影响，包括心率，节拍持续时间,在一定浓度范围内的收缩力和心律失常的证据。作为化合物分析的摘录，S-BayK8644治疗显示心房hiPSC-CM搏动持续时间诱导的浓度依赖性瞬时增加，而心室细胞随着时间的推移显示出搏动速率的生理增加。卡巴胆碱治疗对心房细胞产生显著影响，例如，随着时间的推移，节拍持续时间增加，节拍率下降，但对心室心肌细胞的影响很小。在这种特定于房间的药理学的背景下，我们不仅增加了hiPSC-CM的收缩特性，还提出了一种用于中等通量早期化合物筛选的多孔平台。总的来说,这些见解说明了腔室特异性心肌细胞之间的关键药理学差异及其在多孔收缩性平台上的应用，以获得对体外心脏负债研究和疾病建模的见解。
