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 5

Src/c-Abl pathway participates in integrin α5β1–mediated YAP phosphorylation at Tyr357.

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Src/c-Abl pathway participates in integrin α5β1–mediated YAP phosphoryla...
(A–C) HUVECs were seeded on dishes coated with fibronectin or collagen and cultured for 6 hours with or without inhibitors (10 μmol/l), as indicated. (A and B) Western blot analysis of expression of indicated proteins and ratio of p-YAPY357 to t-YAP. Data are mean ± SEM, *P < 0.05 (2-way ANOVA with Bonferroni multiple comparison post hoc test), n = 6. (C) Immunofluorescence staining of YAP (red) and DAPI (blue). Represent images are shown, n = 6. Scale bars: 20 μm. (D and E) HUVECs were exposed to OSS for 6 hours with or without bosutinib (10 μmol/l). (D) Western blot analysis of p-YAPY357, p-c-AblY412, t-YAP, c-Abl, and VCAM-1. (E) Quantification of protein levels in panel D. Data are mean ± SEM, *P < 0.05 (2-way ANOVA with Bonferroni multiple comparison post hoc test), n = 5. (F) HUVECs were transfected with control or ABL1 siRNA for 24 hours, then underwent OSS or not for 6 hours. Western blot analysis of c-Abl, p-YAPY357, t-YAP, and VCAM-1. (G) Quantification of protein levels in panel F. Data are mean ± SEM, *P < 0.05 (2-way ANOVA with Bonferroni multiple comparison post hoc test), n = 6.