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

Abstract

Chronic cardiac hypertrophy is maladaptive and contributes to the pathogenesis of heart failure. The objective of this study was to identify small molecule inhibitors of pathological cardiomyocyte hypertrophy. High content screening was performed with primary neonatal rat ventricular myocytes (NRVMs) cultured on 96-well plates and treated with a library of 3241 distinct small molecules. Non-toxic hit compounds that blocked hypertrophy in response to phenylephrine (PE) and phorbol myristate acetate (PMA) were identified based on their ability to reduce cell size and inhibit expression of atrial natriuretic factor (ANF), which is a biomarker of pathological cardiac hypertrophy. Many of the hit compounds are existing drugs that have not previously been evaluated for benefit in the setting of cardiovascular disease. One such compound, the anti-malarial drug artesunate, blocked left ventricular hypertrophy (LVH) and improved cardiac function in adult mice subjected to transverse aortic constriction (TAC). These findings demonstrate that phenotypic screening with primary cardiomyocytes can be used to discover anti-hypertrophic lead compounds for heart failure drug discovery. Using annotated libraries of compounds with known selectivity profiles, this screening methodology also facilitates chemical biological dissection of signaling networks that control pathological growth of the heart.

Epigenomic regulation of heart failure: integrating histone marks, long noncoding RNAs, and chromatin architecture

“…Exhaustive and sophisticated medicinal chemistry programs in industry and academia have led to the development of highly selective and potent inhibitors of a wide array of epigenetic targets, and many of the compounds are available to the scientific community through programs such as the Structural Genomics Consortium 30 . Coupling the use of these compounds with well-validated phenotypic assays, such as cell-based assays of cardiomyocyte hypertrophy or fibrosis 31 , has the potential to rapidly uncover novel roles for epigenetic regulators in the control of heart failure and thus provide crucial mechanistic insights.…”

Biology of the cardiac myocyte in heart disease

“…, 2014 ), endothelin-1 (ET-1; Menaouar et al. , 2014 ), and the diacylglycerol mimetic phorbol 12-myristate 12-acetate (PMA; Reid et al. , 2016 ) (see Table 2 later in this paper).…”

“…, 2012 ). Further expansion of this high-throughput imaging approach was used in a recent study that measured cell size and ANF induction in NRVMs exposed to PE and PMA ( Reid et al. , 2016 ).…”

“…, 2016 ). Using this high-content imaging approach, the authors were able to successfully identify compounds that successfully blocked two distinct NRVM hypertrophic models ( Reid et al. , 2016 ).…”

Food Bioactive HDAC Inhibitors in the Epigenetic Regulation of Heart Failure

“…Trichostatin A (TSA), for example, is a pan-HDAC inhibitor that has been shown to inhibit pathological cardiac hypertrophy and fibrosis [ 47 ]. While TSA has been shown to regulate histone hyper-acetylation and gene expression [ 48 , 49 ], its actions on pathological heart enlargement appear to be regulated, in part, through inhibition of mitogen-activated protein kinase (MAPK) signaling [ 50 ]. These data would suggest epigenetic and non-epigenetic (e.g., signaling mediated) mechanisms of action.…”