Phenotypic Screening with Human iPS Cell–Derived Cardiomyocytes: HTS-Compatible Assays for Interrogating Cardiac Hypertrophy

Abstract

A major hurdle for cardiovascular disease researchers has been the lack of robust and physiologically relevant cell-based assays for drug discovery. Derivation of cardiomyocytes from human-induced pluripotent stem (iPS) cells at high purity, quality, and quantity enables the development of relevant models of human cardiac disease with source material that meets the demands of high-throughput screening (HTS). Here we demonstrate the utility of iPS cell-derived cardiomyocytes as an in vitro model of cardiac hypertrophy. Exposure of cardiomyocytes to endothelin 1 (ET-1) leads to reactivation of fetal genes, increased cell size, and robust expression of B-type natriuretic peptide (BNP). Using this system, we developed a suite of assays focused on BNP detection, most notably a high-content imaging-based assay designed for phenotypic screening. Miniaturization of this assay to a 384-well format enabled the profiling of a small set of tool compounds known to modulate the hypertrophic response. The assays described here provide consistent and reliable results and have the potential to increase our understanding of the many mechanisms underlying this complex cardiac condition. Moreover, the HTS-compatible workflow allows for the incorporation of human biology into early phases of drug discovery and development.

Defined Engineered Human Myocardium With Advanced Maturation for Applications in Heart Failure Modeling and Repair

“…To recapitulate sympathetic overstimulation, we exposed EHM to NE at clinically relevant concentrations (0.001–1 μmol/L) for 7 days. We also included a group of EHM exposed to endothelin-1 (0.01 μmol/l), a well established inducer of cardiomyocyte hypertrophy via the alternative Gq-protein transduction pathway 41 . Similarly as observed in patients, chronic NE stimulation induced contractile dysfunction in a concentration dependent manner (Figure 4a) with desensitization to acute β-adrenergic stimulation (Figure 4b), which according to its underlying mechanism only occurred under NE and not ET-1.…”

BET bromodomain inhibition suppresses innate inflammatory and profibrotic transcriptional networks in heart failure

“…We therefore asked whether BET bromodomain inhibition could block pathologic cardiomyocyte hypertrophy in human cells. We tested the effects of JQ1 in a widely used human induced pluripotent stem cell–derived cardiomyocyte (iPSC-CM) culture system that originated from an iPSC line derived from a healthy human donor [iCell Cardiomyocytes, Cellular Dynamics International Inc. (CDI)] (48). Previous studies have demonstrated that these cells can mount a hypertrophic response to 10 nM endothelin-1 (ET-1) (48).…”

“…6C) and other typical marker genes induced in human iPSC-CMs during ET-1 stimulation (fig. S5A) (48). Quantitative enzyme-linked immunosorbent assay (ELISA) confirmed that JQ1 attenuated the ET-1–stimulated increase in BNP protein abundance (Fig.…”

Discovery of novel small molecule inhibitors of cardiac hypertrophy using high throughput, high content imaging

“…Adaptation of this hypertrophy assay to 384-well plates will facilitate expanded screening efforts. Furthermore, this approach could be employed with iPS-derived cardiac myocytes [21], although optimized culture methods will likely be needed to maximize the responses of these cells to hypertrophic agonists [22]. …”