In Vitro Cardiotoxicity Investigation Using High Content Analysis and Human Stem Cell-Derived Models

Abstract

In the quest for more effective approaches to reduce the time, cost, and risks associated with development of new pharmaceuticals, High Content Analysis (HCA) is emerging as a valuable tool for in vitro predictive toxicity testing. Enabling rapid collection and analysis of information-rich cell images from thousands of samples a day, HCA offers high enough throughput for routine screening (High Content Screening, HCS) and mechanistic investigations to be performed at the early stages of discovery and preclinical development. By interrogating multiple endpoints simultaneously in the context of the same cells, candidate drugs can be grouped and prioritized according to the types and severity of cell effects they induce relative to reference compounds. At the same time, the signature drug response profiles captured by HCA provide valuable insights into mechanisms of toxicity. This chapter provides step-by-step protocols used for HCA cardiotoxicity testing of kinase inhibitors and other compounds of interest in oncology. Critical to the success of this approach is the selection and careful handling of a cell model that is amenable to high throughput HCA techniques while faithfully recapitulating human cardiomyocyte physiology in the dish with minimal batch-to-batch variability. Commercial human stem cell-derived cardiomyocytes (hSC-CM) provide a tractable and highly relevant model for this purpose. With robust quality-controlled hSC-CM cultures serving as a common test system, cardiotoxicity data obtained by HCA has the potential to be integrated with data from complementary in vitro tests—particularly those assessing electrophysiological effects—to provide more comprehensive evaluation and improved prediction of clinical responses.