c-Abl regulates YAPY357 phosphorylation to activate endothelial atherogenic responses to disturbed flow
Bochuan Li, … , Yi Zhu, Ding Ai
Bochuan Li, … , Yi Zhu, Ding Ai
Published January 10, 2019
Citation Information: J Clin Invest. 2019;129(3):1167-1179. https://doi.org/10.1172/JCI122440.
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Research Article Vascular biology

c-Abl regulates YAPY357 phosphorylation to activate endothelial atherogenic responses to disturbed flow

Abstract

Local flow patterns determine the uneven distribution of atherosclerotic lesions. This research aims to elucidate the mechanism of regulation of nuclear translocation of Yes-associated protein (YAP) under oscillatory shear stress (OSS) in the atheroprone phenotype of endothelial cells (ECs). We report here that OSS led to tyrosine phosphorylation and strong, continuous nuclear translocation of YAP in ECs that is dependent on integrin α5β1 activation. YAP overexpression in ECs blunted the anti-atheroprone effect of an integrin α5β1–blocking peptide (ATN161) in Apoe–/– mice. Activation of integrin α5β1 induced tyrosine, but not serine, phosphorylation of YAP in ECs. Blockage of integrin α5β1 with ATN161 abolished the phosphorylation of YAP at Y357 induced by OSS. Mechanistic studies showed that c-Abl inhibitor attenuated the integrin α5β1–induced YAP tyrosine phosphorylation. Furthermore, the phosphorylation of c-Abl and YAPY357 was significantly increased in ECs in atherosclerotic vessels of mice and in human plaques versus normal vessels. Finally, bosutinib, a tyrosine kinase inhibitor, markedly reduced the level of YAPY357 and the development of atherosclerosis in Apoe–/– mice. The c-Abl/YAPY357 pathway serves as a mechanism for the activation of integrin α5β1 and the atherogenic phenotype of ECs in response to OSS, and provides a potential therapeutic strategy for atherogenesis.

Authors

Bochuan Li, Jinlong He, Huizhen Lv, Yajin Liu, Xue Lv, Chenghu Zhang, Yi Zhu, Ding Ai

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Figure 3

OSS induced phosphorylation of YAP at Tyr357 through integrin α5β1.

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OSS induced phosphorylation of YAP at Tyr357 through integrin α5β1. (A a...
(A and B) HUVECs were exposed to OSS for different times, as indicated. Cells with static treatment (ST) were a control. Western blot analysis of protein levels of activated integrin α5 (Act-α5), total integrin α5 (t-α5), p-YAPS127, p-YAPS397, p-YAPY357 and t-YAP. (B) Quantification of YAP phosphorylation and Act-α5 normalized with t-YAP and t-α5. Data are mean ± SEM, *P < 0.05 versus ST (1-way ANOVA with Bonferroni multiple comparison post hoc test), n = 5. (C and D) HUVECs were seeded on dishes coated with collagen (Col) or doses of fibronectin (FN) and cultured for 6 hours. Western blot analysis of protein level of activated integrin α5 (Act-α5), total integrin α5 (t-α5), p-YAPS127, p-YAPS397, p-YAPY357, and t-YAP. (D) Quantification of YAP phosphorylation and Act-α5 normalized to t-YAP and t-α5. Data are mean ± SEM, *P < 0.05 (1-way ANOVA with Bonferroni multiple comparison post hoc test), n = 5. (E) HUVECs were exposed to OSS or ST for 6 hours with or without pretreatment of ATN161 (10 μmol/l). Western blot analysis of activated integrin α5 (Act-α5), integrin α5 (t-α5), p-YAPS127, p-YAPS397, p-YAPY357, t-YAP, ICAM-1, and VCAM-1. (F) Quantification of expressions of indicated proteins in panel E. Data are mean ± SEM, *P < 0.05 (2-way ANOVA with Bonferroni multiple comparison post hoc test), n = 6.