Cell-based two-dimensional morphological assessment system to predict cancer drug-induced cardiotoxicity using human induced pluripotent stem cell-derived cardiomyocytes

• Published in: Toxicology and applied pharmacology Vol. 383; p. 114761
• Main Authors: Matsui, Toshikatsu, Miyamoto, Kazumasa, Yamanaka, Kazunori, Okai, Yoshiko, Kaushik, Emily Pfeiffer, Harada, Kousuke, Wagoner, Matthew, Shinozawa, Tadahiro
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 15.11.2019
• Subjects: Antineoplastic Agents - toxicity; Cardiotoxicity; Cardiotoxicity - pathology; Cardiotoxins - toxicity; Cell Culture Techniques - methods; Cells, Cultured; Fluorescent Dyes - analysis; Forecasting; High-content analysis; Humans; Induced pluripotent stem cell-derived cardiomyocytes; Induced Pluripotent Stem Cells - chemistry; Induced Pluripotent Stem Cells - drug effects; Induced Pluripotent Stem Cells - pathology; Microscopy, Electron - methods; Myocytes, Cardiac - chemistry; Myocytes, Cardiac - drug effects; Myocytes, Cardiac - pathology; Sarcomere disruption; Cardiotoxicity; High-content analysis; Induced pluripotent stem cell-derived cardiomyocytes; Sarcomere disruption
• ISSN: 0041-008X; 1096-0333
• DOI: 10.1016/j.taap.2019.114761

Recent developments of novel targeted therapies are contributing to the increased long-term survival of cancer patients; however, drug-induced cardiotoxicity induced by cancer drugs remains a serious problem in clinical settings. Nevertheless, there are few in vitro cell-based assays available to predict this toxicity, especially from the aspect of morphology. Here, we developed a simple two-dimensional (2D) morphological assessment system, 2DMA, to predict drug-induced cardiotoxicity in cancer patients using human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) with image-based high-content analysis in a high-throughput manner. To assess the effects of drugs on cardiomyocytes, we treated iPSC-CMs with 28 marketed pharmaceuticals and measured two key parameters: number of cell nuclei and sarcomere morphology. Drugs that significantly perturbed these two parameters at concentrations ≤30 times the human Cmax value were regarded as positive in the test. Based on these criteria, the sensitivity and specificity of the 2DMA system were 81% and 100%, respectively. Moreover, the translational predictability of 2DMA was comparable with that of a three-dimensional cardiotoxicity assay. RNA sequencing further revealed that the expression levels of several genes related to sarcomere components decreased following treatment with sunitinib, suggesting that inhibition of the synthesis of proteins that comprise the sarcomere contributes to drug-induced sarcomere disruption. Based on these features, the 2DMA system provides mechanistic insight with high predictability of cancer drug-induced cardiotoxicity in humans, and could thus contribute to the reduction of drug attrition rates at early stages of drug development. •Drug-induced change in number of cell nuclei and sarcomere morphology was evaluated.•This assay showed 81% sensitivity and 100% specificity using 28 approved drugs.•Predictability of the assay was comparable with 3D structural cardiotoxicity assay.•Most of the sarcomere genes were downregulated upon treatment with sunitinib.•This assay can help accurately predict cancer drug-induced cardiotoxicity.