Research ArticlePulmonary Hypertension

iPSC–endothelial cell phenotypic drug screening and in silico analyses identify tyrphostin-AG1296 for pulmonary arterial hypertension

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Science Translational Medicine  05 May 2021:
Vol. 13, Issue 592, eaba6480
DOI: 10.1126/scitranslmed.aba6480

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Pursuing promising candidates for PAH

Endothelial cells and smooth muscle cells are dysfunctional in pulmonary arterial hypertension (PAH). Gu et al. used transcriptomics and induced pluripotent stem cell–derived endothelial cells from patients with PAH to identify a tyrosine kinase inhibitor, tyrphostin-AG1296, that improved survival of cells. AG1296 activated bone morphogenetic protein receptor signaling and limited the proliferation of PAH smooth muscle cells, and its drug signature correlated with an anti-PAH gene signature. Treatment of a rat model of pulmonary hypertension reversed vascular remodeling and AG1296-induced regression of lesions in lung organ cultures ex vivo. Combining transcriptomics with functional cell assays could accelerate identification of drug candidates for PAH.

Abstract

Pulmonary arterial hypertension (PAH) is a progressive disorder leading to occlusive vascular remodeling. Current PAH therapies improve quality of life but do not reverse structural abnormalities in the pulmonary vasculature. Here, we used high-throughput drug screening combined with in silico analyses of existing transcriptomic datasets to identify a promising lead compound to reverse PAH. Induced pluripotent stem cell–derived endothelial cells generated from six patients with PAH were exposed to 4500 compounds and assayed for improved cell survival after serum withdrawal using a chemiluminescent caspase assay. Subsequent validation of caspase activity and improved angiogenesis combined with data analyses using the Gene Expression Omnibus and Library of Integrated Network-Based Cellular Signatures databases revealed that the lead compound AG1296 was positively associated with an anti-PAH gene signature. AG1296 increased abundance of bone morphogenetic protein receptors, downstream signaling, and gene expression and suppressed PAH smooth muscle cell proliferation. AG1296 induced regression of PA neointimal lesions in lung organ culture and PA occlusive changes in the Sugen/hypoxia rat model and reduced right ventricular systolic pressure. Moreover, AG1296 improved vascular function and BMPR2 signaling and showed better correlation with the anti-PAH gene signature than other tyrosine kinase inhibitors. Specifically, AG1296 up-regulated small mothers against decapentaplegic (SMAD) 1/5 coactivators, cAMP response element–binding protein 3 (CREB3), and CREB5: CREB3 induced inhibitor of DNA binding 1 and downstream genes that improved vascular function. Thus, drug discovery for PAH can be accelerated by combining phenotypic screening with in silico analyses of publicly available datasets.

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