High-content phenotypic assay for proliferation of human iPSC-derived cardiomyocytes identifies L-type calcium channels as targets

• Published in: Journal of molecular and cellular cardiology Vol. 127; pp. 204 - 214
• Main Authors: Woo, Laura A., Tkachenko, Svyatoslav, Ding, Mei, Plowright, Alleyn T., Engkvist, Ola, Andersson, Henrik, Drowley, Lauren, Barrett, Ian, Firth, Mike, Akerblad, Peter, Wolf, Matthew J., Bekiranov, Stefan, Brautigan, David L., Wang, Qing-Dong, Saucerman, Jeffrey J.
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.02.2019
• Subjects: Calcium Channels, L-Type - metabolism; Cardiomyocyte proliferation; Cell Proliferation; DNA - biosynthesis; High-content imaging; Human iPSC-derived cardiomyocytes; Humans; Image Processing, Computer-Assisted; Induced Pluripotent Stem Cells - cytology; L-Type calcium channel blockers; Myocytes, Cardiac - cytology; Myocytes, Cardiac - metabolism; Phenotype; Phenotypic screen; Ploidies; Cardiomyocyte proliferation; L-Type calcium channel blockers; High-content imaging; Phenotypic screen; Human iPSC-derived cardiomyocytes
• ISSN: 0022-2828; 1095-8584; 1095-8584
• DOI: 10.1016/j.yjmcc.2018.12.015

Over 5 million people in the United States suffer from heart failure, due to the limited ability to regenerate functional cardiac tissue. One potential therapeutic strategy is to enhance proliferation of resident cardiomyocytes. However, phenotypic screening for therapeutic agents is challenged by the limited ability of conventional markers to discriminate between cardiomyocyte proliferation and endoreplication (e.g. polyploidy and multinucleation). Here, we developed a novel assay that combines automated live-cell microscopy and image processing algorithms to discriminate between proliferation and endoreplication by quantifying changes in the number of nuclei, changes in the number of cells, binucleation, and nuclear DNA content. We applied this assay to further prioritize hits from a primary screen for DNA synthesis, identifying 30 compounds that enhance proliferation of human induced pluripotent stem cell-derived cardiomyocytes. Among the most active compounds from the phenotypic screen are clinically approved L-type calcium channel blockers from multiple chemical classes whose activities were confirmed across different sources of human induced pluripotent stem cell-derived cardiomyocytes. Identification of compounds that stimulate human cardiomyocyte proliferation may provide new therapeutic strategies for heart failure. [Display omitted] •Novel automated hybrid live/fixed assay for cardiomyocyte proliferation.•Assay distinguishes proliferation from multinucleation and polyploidization.•Assay applied for high-content screening of compounds.•L-type calcium channel blockers induce cardiomyocyte proliferation.