Chronic Cardiotoxicity Assays Using Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes (hiPSC-CMs)

Abstract

Cardiomyocytes (CMs) differentiated from human induced pluripotent stem cells (hiPSCs) are increasingly used in cardiac safety assessment, disease modeling and regenerative medicine. A vast majority of cardiotoxicity studies in the past have tested acute effects of compounds and drugs; however, these studies lack information on the morphological or physiological responses that may occur after prolonged exposure to a cardiotoxic compound. In this review, we focus on recent advances in chronic cardiotoxicity assays using hiPSC-CMs. We summarize recently published literature on hiPSC-CMs assays applied to chronic cardiotoxicity induced by anticancer agents, as well as non-cancer classes of drugs, including antibiotics, anti-hepatitis C virus (HCV) and antidiabetic drugs. We then review publications on the implementation of hiPSC-CMs-based assays to investigate the effects of non-pharmaceutical cardiotoxicants, such as environmental chemicals or chronic alcohol consumption. We also highlight studies demonstrating the chronic effects of smoking and implementation of hiPSC-CMs to perform genomic screens and metabolomics-based biomarker assay development. The acceptance and wide implementation of hiPSC-CMs-based assays for chronic cardiotoxicity assessment will require multi-site standardization of assay protocols, chronic cardiac maturity marker reproducibility, time points optimization, minimal cellular variation (commercial vs. lab reprogrammed), stringent and matched controls and close clinical setting resemblance. A comprehensive investigation of long-term repeated exposure-induced effects on both the structure and function of cardiomyocytes can provide mechanistic insights and recapitulate drug and environmental cardiotoxicity.

Fluorescent hiPSC-derived MYH6-mScarlet cardiomyocytes for real-time tracking, imaging, and cardiotoxicity assays

“…Following the treatment with different cardiotoxic drugs, we observed a dosedependent effect on CM viability and changes in myocardial architecture. Over the past decade, many studies have shown the use of iPSC-CMs to assess the cardiac toxicity of different drugs (Narkar et al 2022;Sharma et al 2017;Thomas et al 2021). However, monitoring live CMs at different drug concentrations gives an additional application of the reporter CMs on drug dose effects and identifies drugs that interfere with the CMs contractile machinery but may not directly impact the CMs electrophysiology and viability.…”

Human in vitro neurocardiac coculture ( iv NCC ) assay development for evaluating cardiac contractility modulation

“…How to cite this article: Narkar, A., Feaster, T. K., Casciola, M., & Blinova, K. (2022). Human in vitro neurocardiac coculture (ivNCC) assay development for evaluating cardiac contractility modulation.…”

“…Primary and secondary antibodies were diluted in 2.5% (wt/vol) bovine serum albumin (BSA)/PBS/0.5% Triton X-100. Specimens were incubated with primary antibodies Cardiac troponin (Catalog # MA5-12960, Invitrogen), Peripherin (Catalog # AB1530, EMD Millipore) at 1:500 overnight, washed three times for 5-10 min, and incubated with fluorescently conjugated secondary antibodies for 2-4 h as described in detail elsewhere (Narkar et al, 2021). All washes were performed with PBS/0.5% Triton X-100.…”

“…The tridiagonal medical treatment strategy for systolic heart failure includes, but is not limited to, β‐blockers, angiotensin‐converting enzyme inhibitors, and mineralocorticoid receptor antagonists. Recent additions include sacubitril/valsartan and ivabradine for heart failure (Campbell et al, 2020 ; Narkar et al, 2022 ; Upadhya & Kitzman, 2020 ). Recently added to the treatment regimen is cardiac contractility modulation (CCM), an FDA approved intracardiac device therapy that targets patients with chronic heart failure, QRS <130 ms and a left ventricular ejection fraction (LVEF) 25%–45% who are currently receiving optimal medical therapy (OMT) yet remain symptomatic (Campbell et al, 2020 ; FDA.gov, 2019 ).…”

Chronic cardiotoxicity assessment by cell optoporation on microelectrode arrays

“…Although hiPSC-CMs can be maintained in culture for long periods of time with stable baseline functions (Kopljar et al 2017;Dias et al 2018), the current methods such as patch-clamp, voltage indicators, and MEA technology do not offer at the same time high sensitivity for measuring transmembrane ion currents and low-invasiveness for 3 monitoring cells over long time. Therefore, despite the recent advances in chronic cardiotoxicity assays using hiPSC-CMs (Narkar et al 2022), long-term (chronic) cardiac adverse effects remain a major barrier of drug development. The in vitro detection of chronic cardiotoxicity represents a key turning point to avoid the progression of a drug candidate with "delayed" cardiac adverse effect.…”