Isorhapontigenin protects against doxorubicin-induced cardiotoxicity via increasing YAP1 expression

Abstract

As an effective anticancer drug, the clinical limitation of doxorubicin (Dox) is the time- and dose-dependent cardiotoxicity. Yes-associated protein 1 (YAP1) interacts with transcription factor TEA domain 1 (TEAD1) and plays an important role in cell proliferation and survival. However, the role of YAP1 in Dox-induced cardiomyopathy has not been reported. In this study, the expression of YAP1 was reduced in clinical human failing hearts with dilated cardiomyopathy and Dox-induced in vivo and in vitro cardiotoxic model. Ectopic expression of Yap1 significantly blocked Dox-induced cardiomyocytes apoptosis in TEAD1 dependent manner. Isorhapontigenin (Isor) is a new derivative of stilbene and responsible for a wide range of biological processes. Here, we found that Isor effectively relieved Dox-induced cardiomyocytes apoptosis in a dose-dependent manner in vitro. Administration with Isor (30 mg/kg/day, intraperitoneally, 3 weeks) significantly protected against Dox-induced cardiotoxicity in mice. Interestingly, Isor increased Dox-caused repression in YAP1 and the expression of its target genes in vivo and in vitro. Knockout or inhibition of Yap1 blocked the protective effects of Isor on Dox-induced cardiotoxicity. In conclusion, YAP1 may be a novel target for Dox-induced cardiotoxicity and Isor might be a new compound to fight against Dox-induced cardiotoxicity by increasing YAP1 expression.

Keywords: AMPK, AMP-activated protein kinase; AP-1, anti-microbial protein; AREG, amphiregulin; AUC/Dose, dose-normalized plasma exposures; Amphiregulin; Ang II, angiotensin II; CO, cardiac output; CTGF, connective tissue growth factor; Cardiomyocytes apoptosis; Cardiotoxicity; Cmax/Dose, dose-normalized maximal plasma concentrations; Connective tissue growth factor; DAB, 3,3′-diaminobenzidine; DMEM, Dulbecco's modified Eagle's medium; Dob, dobutamine; Dox, doxorubicin; Doxorubicin; EMT, epithelial mesenchymal transformation; FOXO1, forkhead box class O1; FS, fractional shortening; HE, hematoxylin–eosin; ISO, isoproterenol; Isor, isorhapontigenin; Isorhapontigenin; LVAW;d, left ventricular end-diastolic anterior wall thickness; LVAW;s, left ventricular end-systolic anterior wall thickness; LVEF, left ventricular ejection fraction; LVID;d, left ventricular end-diastolic internal diameter; LVID;s, left ventricular end-systolic internal diameter; LVPW;d, left ventricular end-diastolic posterior wall thickness; LVPW;s, left ventricular end-systolic posterior wall thickness; MAPK, mitogen-activated protein kinase; MI, myocardial infarction; NF-κB, nuclear factor kappa-B; NRCMs, neonatal rat cardiomyocytes; P2Y12 receptor, ADP receptor; PGC-1α, peroxisome proliferator-activated receptor γ coactivator-1α; PMSF, phenylmethanesulfonyl fluoride; PVDF, polyvinylidene fluoride; ROS, reactive oxygen species; SD, Sprague–Dawley; SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis; SESN2, sestrin2; TCF4, T-cell factor 4; TEAD, TEA domain transcription factor proteins; TEAD1; TUNEL, TdT-mediated dUTP nick end labeling; WGA, wheat germ agglutinin; YAP1; YAP1, Yes-associated protein 1; qRT-PCR, quantitative real-time polymerase chain reaction; sgRNAs, sequence guiding RNAs; Δψm, mitochondrial membrane potential.