Phenotypic screening with human iPS cell-derived cardiomyocytes: HTS-compatible assays for interrogating cardiac hypertrophy.
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.
1Cellular Dynamics International, Madison, WI, USA.
Coby
C
Carlson
Chad
C
Koonce
Natsuyo
N
Aoyama
Shannon
S
Einhorn
Steve
S
Fiene
Arne
A
Thompson
Brad
B
Swanson
Blake
B
Anson
Steven
S
Kattman
D015415:Biomarkers / Q000378:metabolism; D002121:Calcium Channel Blockers / Q000494:pharmacology; D006332:Cardiomegaly / Q000188:drug therapy; D002454:Cell Differentiation; D048429:Cell Size; D002478:Cells, Cultured; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D015870:Gene Expression; D057166:High-Throughput Screening Assays; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D057026:Induced Pluripotent Stem Cells / Q000502:physiology*; D020128:Inhibitory Concentration 50; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000378:metabolism; D010641:Phenotype; D011804:Quinolines / Q000494:pharmacology; D017461:Receptors, Atrial Natriuretic Factor / Q000235:genetics / Q000378:metabolism; D014700:Verapamil / Q000494:pharmacology; D000099242:Atrial Natriuretic Factor Receptor B
D015415:Biomarkers; D002121:Calcium Channel Blockers; D056572:Histone Deacetylase Inhibitors; D007093:Imidazoles; D011804:Quinolines; D017461:Receptors, Atrial Natriuretic Factor; D014700:Verapamil; D000099242:Atrial Natriuretic Factor Receptor B; C531198:dactolisib
cardiomyocytes; hypertrophy; iPS cell; phenotypic screening