Vinculin phosphorylation impairs vascular endothelial junctions promoting atherosclerosis. (16th November 2022)

• Record Type: Journal Article
• Authors: Shih, Yu-Tsung; Wei, Shu-Yi; Chen, Jin-Hua; Wang, Wei-Li; Wu, Hsin-Yi; Wang, Mei-Cun; Lin, Chia-Yu; Lee, Pei-Lin; Lin, Chih-Yuan; Chiang, Hung-Che; Chen, Yu-Ju; Chien, Shu; Chiu, Jeng-Jiann
• Is Part Of: European heart journal. Volume 44:Number 4(2023)
• Page Start: 304
• Page End: 318
• Publication Date: 2022-11-16
• Subjects: Atherosclerosis -- Disturbed flow -- Endothelial cell -- Phosphoproteomics -- Shear stress -- Vinculin; Cardiology -- Periodicals; Heart -- Diseases -- Periodicals; 616.12005
• DOI: 10.1093/eurheartj/ehac647
• Languages: English
• ISSNs: 0195-668X

Abstract

Abstract: Background and aims: Atherosclerosis preferentially develops in arterial branches and curvatures where vascular endothelium is exposed to disturbed flow. In this study, the effects of disturbed flow on the regulation of vascular endothelial phosphoproteins and their contribution to therapeutic application in atherogenesis were elucidated. Methods: Porcine models, large-scale phosphoproteomics, transgenic mice, and clinical specimens were used to discover novel site-specific phosphorylation alterations induced by disturbed flow in endothelial cells (ECs). Results: A large-scale phosphoproteomics analysis of native endothelium from disturbed (athero-susceptible) vs. pulsatile flow (athero-resistant) regions of porcine aortas led to the identification of a novel atherosclerosis-related phosphoprotein vinculin (VCL) with disturbed flow-induced phosphorylation at serine 721 (VCL S721p ). The induction of VCL S721p was mediated by G-protein-coupled receptor kinase 2 (GRK2) S29p and resulted in an inactive form of VCL with a closed conformation, leading to the VE-cadherin/catenin complex disruption to enhance endothelial permeability and atherogenesis. The generation of novel apolipoprotein E-deficient ( ApoE −/− ) mice overexpressing S721-non-phosphorylatable VCL mutant in ECs confirmed the critical role of VCL S721p in promoting atherosclerosis. The administration of a GRK2 inhibitor to ApoE −/− mice suppressed plaque formation by inhibiting endothelial VCL S721p . Studies on clinical specimens from patients with coronary artery disease (CAD) revealed that endothelial VCL S721p is a critical clinicopathological biomarker for atherosclerosis progression and that serum VCL S721p level is a promising biomarker for CAD diagnosis. Conclusions: The findings of this study indicate that endothelial VCL S721p is a valuable hemodynamic-based target for clinical assessment and treatment of vascular disorders resulting from atherosclerosis. Structured Graphical Abstract: Structured Graphical Abstract Proposed mechanisms by which disturbed flow induces vinculin phosphorylation at serine 721 (VCL S721p ) in vascular endothelium, leading to atherosclerosis. A combination of porcine models, large-scale phosphoproteomics, transgenic mice, and clinical specimens was used to demonstrate that disturbed flow induces endothelial VCL S721p via G-protein-coupled receptor kinase 2 (GRK2) S29p, resulting in an inactive form of VCL with a closed conformation. This disrupted the VE-cadherin junction/catenin complex to enhance endothelial permeability and atherosclerosis. Statin therapy was associated with reduced levels of sVCL S721p and sVCL S721p /sVCL in the serum of patients with coronary artery disease. Thus, endothelial VCL S721p and serum levels of sVCL S721p and sVCL S721p /sVCL are promising molecular targets for the clinical assessment and treatment of atherosclerosis. AJ, adherens junction.