Shu-Yi Wei1, Yu-Tsung Shih1, Hsin-Yi Wu2, Wei-Li Wang1, Pei-Ling Lee1, Chih-I Lee1, Chia-Yu Lin1, Yu-Ju Chen3, Shu Chien4, Jeng-Jiann Chiu1,5,6,7,8,9. 1. Institute of Cellular and System Medicine, National Health Research Institutes, Miaoli, Taiwan (S.-Y.W., Y.-T.S., W.-L.W., P.-L.L., C.-I.L., C.-Y.L., J.-J.C.). 2. Instrumentation Center (H.-Y.W.), National Taiwan University, Taipei, Taiwan. 3. Institute of Chemistry, Academic Sinica, Taipei, Taiwan (Y.-J.C.). 4. Department of Bioengineering and Medicine, and Institute of Engineering in Medicine, University of California, San Diego, La Jolla (S.C.). 5. Institute of Polymer Science and Engineering (J.-J.C.), National Taiwan University, Taipei, Taiwan. 6. School of Medical Laboratory Science and Biotechnology, College of Medical Science and Technology (J.-J.C.), Taipei Medical University, Taiwan. 7. Ph.D. Program in Medical Biotechnology, College of Medical Science and Technology (J.-J.C.), Taipei Medical University, Taiwan. 8. Taipei Heart Institute (J.-J.C.), Taipei Medical University, Taiwan. 9. Institute of Biomedical Engineering, National Tsing Hua University, Hsinchu, Taiwan (J.-J.C.).
Abstract
RATIONALE: Disturbed flow occurring in arterial branches and curvatures induces vascular endothelial cell (EC) dysfunction and atherosclerosis. We postulated that disturbed flow plays important role in modulating phosphoprotein expression profiles to regulate endothelial functions and atherogenesis. OBJECTIVE: The goal of this study is to discover novel site-specific phosphorylation alterations induced by disturbed flow in ECs to contribute to atherosclerosis. METHODS AND RESULTS: Quantitative phosphoproteomics analysis of ECs exposed to disturbed flow with low and oscillatory shear stress (0.5±4 dynes/cm2) versus pulsatile shear stress (12±4 dynes/cm2) revealed that oscillatory shear stress induces phospho-YY1S118 (serine [S]118 phosphorylation of Yin Yang 1) in ECs. Elevated phospho-YY1S118 level in ECs was further confirmed to be present in the disturbed flow regions in experimental animals and human atherosclerotic arteries. This disturbed flow-induced EC phospho-YY1S118 is mediated by CK2α (casein kinase 2α) through its direct interaction with YY1. Yeast 2-hybrid library screening and in situ proximity ligation assays demonstrate that phospho-YY1S118 directly binds ZKSCAN4 (zinc finger with KRAB [krüppel-associated box] and SCAN [SRE-ZBP, CTfin51, AW-1 and Number 18 cDNA] domains 4) to induce promoter activity and gene expression of HDM2 (human double minute 2), which consequently induces EC proliferation through downregulation of p53 and p21CIP1. Administration of apoE-deficient (ApoE-/-) mice with CK2-specific inhibitor tetrabromocinnamic acid or atorvastatin inhibits atherosclerosis formation through downregulations of EC phospho-YY1S118 and HDM2. Generation of novel transgenic mice bearing EC-specific overexpression of S118-nonphosphorylatable mutant of YY1 in ApoE-/- mice confirms the critical role of phospho-YY1S118 in promoting atherosclerosis through EC HDM2. CONCLUSIONS: Our findings provide new insights into the mechanisms by which disturbed flow induces endothelial phospho-YY1S118 to promote atherosclerosis, thus indicating phospho-YY1S118 as a potential molecular target for atherosclerosis treatment.
RATIONALE: Disturbed flow occurring in arterial branches and curvatures induces vascular endothelial cell (EC) dysfunction and atherosclerosis. We postulated that disturbed flow plays important role in modulating phosphoprotein expression profiles to regulate endothelial functions and atherogenesis. OBJECTIVE: The goal of this study is to discover novel site-specific phosphorylation alterations induced by disturbed flow in ECs to contribute to atherosclerosis. METHODS AND RESULTS: Quantitative phosphoproteomics analysis of ECs exposed to disturbed flow with low and oscillatory shear stress (0.5±4 dynes/cm2) versus pulsatile shear stress (12±4 dynes/cm2) revealed that oscillatory shear stress induces phospho-YY1S118 (serine [S]118 phosphorylation of Yin Yang 1) in ECs. Elevated phospho-YY1S118 level in ECs was further confirmed to be present in the disturbed flow regions in experimental animals and human atherosclerotic arteries. This disturbed flow-induced EC phospho-YY1S118 is mediated by CK2α (casein kinase 2α) through its direct interaction with YY1. Yeast 2-hybrid library screening and in situ proximity ligation assays demonstrate that phospho-YY1S118 directly binds ZKSCAN4 (zinc finger with KRAB [krüppel-associated box] and SCAN [SRE-ZBP, CTfin51, AW-1 and Number 18 cDNA] domains 4) to induce promoter activity and gene expression of HDM2 (human double minute 2), which consequently induces EC proliferation through downregulation of p53 and p21CIP1. Administration of apoE-deficient (ApoE-/-) mice with CK2-specific inhibitor tetrabromocinnamic acid or atorvastatin inhibits atherosclerosis formation through downregulations of EC phospho-YY1S118 and HDM2. Generation of novel transgenic mice bearing EC-specific overexpression of S118-nonphosphorylatable mutant of YY1 in ApoE-/- mice confirms the critical role of phospho-YY1S118 in promoting atherosclerosis through EC HDM2. CONCLUSIONS: Our findings provide new insights into the mechanisms by which disturbed flow induces endothelial phospho-YY1S118 to promote atherosclerosis, thus indicating phospho-YY1S118 as a potential molecular target for atherosclerosis treatment.
Authors: Celine Souilhol; Blanca Tardajos Ayllon; Xiuying Li; Mannekomba R Diagbouga; Ziqi Zhou; Lindsay Canham; Hannah Roddie; Daniela Pirri; Emily V Chambers; Mark J Dunning; Mark Ariaans; Jin Li; Yun Fang; Helle F Jørgensen; Michael Simons; Rob Krams; Johannes Waltenberger; Maria Fragiadaki; Victoria Ridger; Sarah De Val; Sheila E Francis; Timothy Ja Chico; Jovana Serbanovic-Canic; Paul C Evans Journal: Sci Adv Date: 2022-08-31 Impact factor: 14.957