High-throughput screening of tyrosine kinase inhibitor cardiotoxicity with human induced pluripotent stem cells

Abstract

Tyrosine kinase inhibitors (TKIs), despite efficacy as anti-cancer therapeutics, are associated with cardiovascular side effects ranging from induced arrhythmias to heart failure. We used patient-specific human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs), generated from 11 healthy individuals and 2 patients receiving cancer treatment, to screen FDA-approved TKIs for cardiotoxicities by measuring alterations in cardiomyocyte viability, contractility, electrophysiology, calcium handling, and signaling. With these data, we generated a “cardiac safety index” to assess cardiotoxicities of existing TKIs. TKIs with low cardiac safety indices exhibit cardiotoxicity in patients. We also derived endothelial cells (hiPSC-ECs) and cardiac fibroblasts (hiPSC-CFs) to examine cell type-specific cardiotoxicities. Using high-throughput screening, we determined that VEGFR2/PDGFR-inhibiting TKIs caused cardiotoxicity in hiPSC-CMs, hiPSC-ECs, and hiPSC-CFs. Using phosphoprotein analysis, we determined that VEGFR2/PDGFR-inhibiting TKIs led to a compensatory increase in cardioprotective insulin and insulin-like growth factor (IGF) signaling in hiPSC-CMs. Upregulating cardioprotective signaling with exogenous insulin or IGF1 improved hiPSC-CM viability during co-treatment with cardiotoxic VEGFR2/PDGFR-inhibiting TKIs. Thus, hiPSC-CMs can be used to screen for cardiovascular toxicities associated with anti-cancer TKIs, correlating with clinical phenotypes. This approach provides unexpected insights, as illustrated by our finding that toxicity can be alleviated via cardioprotective insulin/IGF signaling.

High throughput physiological screening of iPSC-derived cardiomyocytes for drug development

“…The potential exists to correlate drug dosing and plasma concentrations with cardiotoxic and myopathic risk. Recent studies suggest that iPSC-cardiomyocytes can reveal cardiotoxicities manifesting as alterations in cardiomyocyte viability, contractility, intracellular signaling and gene expression [127, 128]. As above, however, there is great need for validation, especially considering that many anti-cancer drugs elicit their cardiac effects by affecting mitochondrial function [129, 130] and that iPSC-cardiomyocytes are metabolically immature [131].…”

Multiscale technologies for treatment of ischemic cardiomyopathy

“…Culturing such CMs is of great interest not only because of their applications in cell therapy but also owing to their considerable potential in the patient-specific study of heart disease 64,65 . Human-iPSCs (hiPSCs) have created unique opportunities in cardiology; specifically, CMs differentiated from these stem cells have excellent potential in the study of heart disease and can model patient-specific myocardial physiology in vitro 47,66,67 . However, the substantial structural and functional differences between hiPSC/MSC-derived CMs (derived from existing chemically defined approaches) and adult mature CMs pose major challenges for precise drug screening and highly efficient therapeutic applications 65,68–71 .…”

Mechanistic Systems Modeling to Improve Understanding and Prediction of Cardiotoxicity Caused by Targeted Cancer Therapeutics

“…Although cardiovascular pharmacology has traditionally not been well-suited for high throughput studies, the development of hiPSC-CMs has expanded the possibilities. For instance, a recent study described the development of a comprehensive assay that evaluated cellular effects of TKIs in hiPSC-CMs (Sharma et al, 2017). Using hiPSC lines from 13 individuals, the investigators examined how 21 FDA-approved small molecule TKIs affected cell viability, contractility, and gene expression.…”

“…Using hiPSC lines from 13 individuals, the investigators examined how 21 FDA-approved small molecule TKIs affected cell viability, contractility, and gene expression. By integrating the results with literature-reported TKI serum levels in patients, the authors developed a novel cardiac safety index for TKIs (Sharma et al, 2017).…”

“…As noted above, an important recent study by Sharma et al (2017) derived a "toxicity index" by examining effects of TKIs in hiPSC-CMs through several assays. Although the large-scale nature of this study justifies the "systems" label, and the toxicity index is a quantitative risk score, it's important to emphasize that QSP modeling, which can provide mechanistic insight and actionable predictions, is complementary to a high-throughput, data-driven strategy such as that used in this study (Sharma et al, 2017).…”