Division of Pediatric Cardiology, Department of Pediatrics, Emory University School of Medicine and Children's Healthcare of Atlanta, Atlanta, Georgia.
Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, Georgia.
Toxicol Sci. 2019 May 1;169(1):280-292. doi: 10.1093/toxsci/kfz038.
Alcohol use prior to and during pregnancy remains a significant societal problem and can lead to developmental fetal abnormalities including compromised myocardia function and increased risk for heart disease later in life. Alcohol-induced cardiac toxicity has traditionally been studied in animal-based models. These models have limitations due to physiological differences from human cardiomyocytes (CMs) and are also not suitable for high-throughput screening. We hypothesized that human-induced pluripotent stem cell-derived CMs (hiPSC-CMs) could serve as a useful tool to study alcohol-induced cardiac defects and/or toxicity. In this study, hiPSC-CMs were treated with ethanol at doses corresponding to the clinically relevant levels of alcohol intoxication. hiPSC-CMs exposed to ethanol showed a dose-dependent increase in cellular damage and decrease in cell viability, corresponding to increased production of reactive oxygen species. Furthermore, ethanol exposure also generated dose-dependent increased irregular Ca2+ transients and contractility in hiPSC-CMs. RNA-seq analysis showed significant alteration in genes belonging to the potassium voltage-gated channel family or solute carrier family, partially explaining the irregular Ca2+ transients and contractility in ethanol-treated hiPSC-CMs. RNA-seq also showed significant upregulation in the expression of genes associated with collagen and extracellular matrix modeling, and downregulation of genes involved in cardiovascular system development and actin filament-based process. These results suggest that hiPSC-CMs can be a novel and physiologically relevant system for the study of alcohol-induced cardiac toxicity.
在怀孕前后饮酒仍然是一个严重的社会问题，可能导致胎儿发育异常，包括心肌功能受损和日后患心脏病的风险增加。酒精引起的心脏毒性传统上是在基于动物的模型中进行研究的。这些模型由于与人类心肌细胞（CMs）的生理差异而存在局限性，并且也不适合高通量筛选。我们假设人诱导多能干细胞衍生的 CMs（hiPSC-CMs）可以作为研究酒精引起的心脏缺陷和/或毒性的有用工具。在这项研究中，将乙醇以对应于临床相关酒精中毒水平的剂量处理 hiPSC-CMs。暴露于乙醇的 hiPSC-CMs 表现出细胞损伤的剂量依赖性增加和细胞活力的降低，这与活性氧的产生增加相对应。此外，乙醇暴露还产生了剂量依赖性增加的不规则 Ca2+瞬变和 hiPSC-CMs 的收缩性。RNA-seq 分析显示，属于钾电压门控通道家族或溶质载体家族的基因发生了显著改变，部分解释了乙醇处理的 hiPSC-CMs 中不规则的 Ca2+瞬变和收缩性。RNA-seq 还显示与胶原蛋白和细胞外基质建模相关的基因表达显著上调，与心血管系统发育和肌动蛋白丝为基础的过程相关的基因表达下调。这些结果表明，hiPSC-CMs 可以成为研究酒精引起的心脏毒性的新型生理相关系统。
