Abstract 18674: High-Content Phenotypic Screen for Compounds that Induce Proliferation of Human iPSC-Derived Cardiomyocytes
Introduction: Over 5 million people in the United States are affected by heart failure, a consequence of the heart’s limited ability to regenerate functional cardiac tissue post-injury. Enhancing proliferation of resident cardiomyocytes is a promising potential therapeutic strategy for cardiac regeneration. Here, we performed high-content proliferation screens to identify compounds that enhance proliferation of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs).
Methods: We performed an initial high-content screen of hiPSC-CMs of 5000 compounds from AstraZeneca’s phenotypic screening library consisting of pre-clinical, clinical, and tool compounds. The primary screen measured DNA synthesis in hiPSC-CMs exposed to EdU over 2 days, and the initial hits were assayed in a counter-screen of cardiac fibroblasts to select compounds that preferentially induced proliferation in hiPSC-CMs. To further prioritize the candidate compounds, we developed a novel secondary screen assay that combined automated live-cell microscopy and image analysis. The secondary screen assay tracked changes in cell counts, DNA content, Ki67, and binucleation events over 6 days. We assayed 5-point concentration responses to the top 48 compounds and implemented automated segmentation and image processing algorithms to extract proliferation metrics.
Results: Quantitative image analysis of the primary screen identified a rank-ordered list of 48 compounds that maximally enhanced DNA synthesis without toxicity in in hiPSC-CMs. Secondary screening identified that 41 of those 48 compounds also enhanced hiPSC-CM proliferation. The top compounds putatively target proteins previously shown (e.g. p38, GSK3, CaMKII) as well as proteins not previously associated with cardiomyocyte proliferation (e.g. L-type calcium channels and dopamine receptors).
Conclusions: This study establishes a high-content hiPSC-CM proliferation assay, as well as compounds and potential new mechanisms that modulate cardiomyocyte proliferation.
Author Disclosures: L.A. Woo: None. S. Tkachenko: None. M. Ding: Employment; Significant; AstraZeneca. A.T. Plowright: Employment; Significant; AstraZeneca. O. Engkvist: Employment; Significant; AstraZeneca. H. Andersson: Employment; Significant; AstraZeneca. L. Drowley: Employment; Significant; AstraZeneca. I. Barrett: Employment; Significant; AstraZeneca. M. Firth: Employment; Significant; AstraZeneca. M.J. Wolf: None. S. Bekiranov: None. D.L. Brautigan: None. Q. Wang: Employment; Significant; AstraZeneca. J.J. Saucerman: Research Grant; Significant; AstraZeneca.
- © 2016 by American Heart Association, Inc.