Cardiac Toxicity From Ethanol Exposure in Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes

Abstract

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 Ca 2þ 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 Ca 2þ 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.

Melphalan induces cardiotoxicity through oxidative stress in cardiomyocytes derived from human induced pluripotent stem cells

“…Differentiation cultures were analyzed for CM purity using antibodies against NKX2-5, a cardiac specific transcription factor. Images were acquired and quantitatively analyzed using ArrayScan TM XTI Live High Content Platform (Thermo Fisher Scientific) with mask modifiers for NKX2-5 restricted to the nucleus [21].…”

“…Ca 2+ transient assay. hiPSC-CMs at low density were stained with 5µM Fluo-4 AM in 1× normal Tyrode solution [21]. Dynamic fluorescent images were recorded using the ImageXpress Micro…”

“…Particularly, SOD3 level detected was 5.5 times higher in cells exposed to 10 µM melphalan compared with no melphalan treatment and even higher To further evaluate if ROS production plays a crucial role in melphalan-induced cardiotoxicity, we treated hiPSC-CMs with 0, 10, and 20 µM melphalan in combination with or without 1 mM of ROS scavenger NAC concomitantly, for 3 days, and measured cell viability and ROS production. The dose selection of NAC was based on previous studies in which 1 mM of NAC effectively attenuated the ethanol-and doxorubicin-induced oxidative stress in hiPSC-CMs [21,27]. As shown in Fig.…”

Beyond Family: Modeling Non-hereditary Heart Diseases With Human Pluripotent Stem Cell-Derived Cardiomyocytes

“…Similarly, hPSC-CMs were used to model the role of the mitochondrial calcium uniporter in cardiac fetal development and maturation (Shanmughapriya et al, 2018). Finally, although chemically induced cardiotoxicity will not be a subject of this review [see (Magdy et al, 2018)], one recent study considered the impact of ethanol on hPSC-CM functionality as a model of prenatal exposure during maternal alcohol intoxication (Rampoldi et al, 2019).…”

Melphalan induces cardiotoxicity through oxidative stress in cardiomyocytes derived from human induced pluripotent stem cells

“…Differentiation cultures were analyzed for CM purity using antibodies against NKX2-5, a cardiac speci c transcription factor. Images were acquired and quantitatively analyzed using ArrayScan XTI Live High Content Platform (Thermo Fisher Scienti c) with mask modi ers for NKX2-5 restricted to the nucleus [22].…”