A predictive in vitro risk assessment platform for pro-arrhythmic toxicity using human 3D cardiac microtissues

Abstract

Cardiotoxicity of pharmaceutical drugs, industrial chemicals, and environmental toxicants can be severe, even life threatening, which necessitates a thorough evaluation of the human response to chemical compounds. Predicting risks for arrhythmia and sudden cardiac death accurately is critical for defining safety profiles. Currently available approaches have limitations including a focus on single select ion channels, the use of non-human species in vitro and in vivo, and limited direct physiological translation. We have advanced the robustness and reproducibility of in vitro platforms for assessing pro-arrhythmic cardiotoxicity using human induced pluripotent stem cell-derived cardiomyocytes and human cardiac fibroblasts in 3-dimensional microtissues. Using automated algorithms and statistical analyses of eight comprehensive evaluation metrics of cardiac action potentials, we demonstrate that tissue-engineered human cardiac microtissues respond appropriately to physiological stimuli and effectively differentiate between high-risk and low-risk compounds exhibiting blockade of the hERG channel (E4031 and ranolazine, respectively). Further, we show that the environmental endocrine disrupting chemical bisphenol-A (BPA) causes acute and sensitive disruption of human action potentials in the nanomolar range. Thus, this novel human 3D in vitro pro-arrhythmic risk assessment platform addresses critical needs in cardiotoxicity testing for both environmental and pharmaceutical compounds and can be leveraged to establish safe human exposure levels.

A Change of Heart: Human Cardiac Tissue Engineering as a Platform for Drug Development

“…It was demonstrated that this platform was able to detect changes in cellular viability, endoplasmic reticulum integrity, and mitochondrial membrane potential. Beyond structural effects, cardiac spheroids have been used to stratify pro-arrhythmic toxicity of hERG channel blockers and environmental toxins [ 34 ]. Using cardiac spheroids, it is also possible to model disease states.…”

Assessing Drug-Induced Mitochondrial Toxicity in Cardiomyocytes: Implications for Preclinical Cardiac Safety Evaluation

“…The reactivity of cardiac organoids induced by clinical compounds, including antibiotic, antidiabetic, and anticancer drugs, was shown to be more physiologically relevant compared with 2D-cultured hiPSC-CMs [ 335 , 336 ]. Based on a panel of eight metrics, cardiac organoids responded appropriately to pro-arrhythmic stimuli and effectively differentiated between high- and low-risk hERG-inhibiting compounds, meeting the critical demand in pro-arrhythmic cardiotoxicity prediction [ 337 ]. In addition to electrophysiology, cardiac organoids are also sensitive to drugs affecting cardiac contractility and can be applied in HTS format using a customized image acquisition workflow and optical flow analysis methods [ 329 , 335 ].…”

Integrating nonlinear analysis and machine learning for human induced pluripotent stem cell‐based drug cardiotoxicity testing

“…Recently, automated algorithms and in-depth statistical analyses have gathered more attention to effectively differentiate the cardiotoxicity types for drug candidates (Kofron et al, 2021). For example, a semi-automatic data analysis software was developed to improve the accuracy, reliability, and throughput of measurements of hiPSC-CMs' field potentials.…”

“…Using commercially available calcium sensitive dyes or genetically encoded fluorescent calcium indicators (e.g., GCaMP6f), intracellular calcium transient can be measured through fluorescent microscopy to represent the contractile functionality of hiPSC‐CMs under drug exposure (Sirenko et al., 2013). Recently, automated algorithms and in‐depth statistical analyses have gathered more attention to effectively differentiate the cardiotoxicity types for drug candidates (Kofron et al., 2021). For example, a semi‐automatic data analysis software was developed to improve the accuracy, reliability, and throughput of measurements of hiPSC‐CMs’ field potentials.…”