This competitive renewal application proposes to investigate the downstream mechanisms by which ROCK regulates eNOS mRNA stability and activity. Preliminary studies suggest that phosphorylation of the translational elongation factor-1? (eEF1A) by ROCK2 leads to decreased eNOS mRNA stability and expression. Furthermore, we found that activation of Phosphatase and TENsin homologue (PTEN) by ROCK2 leads to decreased phosphatidylinositol 3,4,5-trisphosphate (PtdIns-3,4,5-P3) levels and inhibition of Akt-mediated eNOS activity. Based upon these pilot studies, two comprehensive specific aims are proposed, which will determine how the regulation of eEF1A and PTEN by ROCK2 could affect eNOS expression and activity, respectively, and whether this contributes importantly to the pathogenesis of endothelial dysfunction and vascular disease. It is hoped that these studies will help establish the importance of ROCK2 as a therapeutic targets for improving endothelial function and decreasing cardiovascular disease.
Specific aim 1 : To determine whether ROCK2 regulates endothelial function in vivo, we will generate endothelial-specific ROCK2Tie2-/- and EC-caROCK mice, and investigate whether eNOS expression, endothelium-dependent vascular relaxation, and atherosclerosis are altered due to the loss- or gain-of-function of endothelial ROCK2. Further studies are proposed to determine how phosphorylation of eEF1A by ROCK2 leads to eNOS mRNA instability and decreased eNOS expression.
Specific aim 2 : To determine how ROCK2 could inhibit the Akt/eNOS pathway, we will investigate how ROCK2 decreases PIP3 levels through inhibition of PI3K activity, stimulation of PTEN activity, or both. We will determine whether ROCK2 can bind to and phosphorylate PTEN, and identify putative ROCK phosphorylation site(s) on PTEN. We will then determine whether phosphorylation of PTEN by ROCK2 increases PTEN activity, leading to decreased PIP3 levels and inhibition of Akt Thr308 phosphorylation and activity. These findings will be correlated with Akt-dependent eNOS Ser1179 phosphorylation and the degree of vascular remodeling.

Public Health Relevance

Cardiovascular disease is a major cause of morbidity and mortality worldwide. The underlying mechanism is partly due to endothelial dysfunction. This research application proposes to investigate the role of an emerging signaling pathway, Rho kinase (ROCK), as an important regulator of endothelial function in vascular disease.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
2R01HL052233-15
Application #
8439356
Study Section
Vascular Cell and Molecular Biology Study Section (VCMB)
Program Officer
Liu, Lijuan
Project Start
1995-07-01
Project End
2017-06-30
Budget Start
2013-07-17
Budget End
2014-06-30
Support Year
15
Fiscal Year
2013
Total Cost
$376,040
Indirect Cost
$138,040
Name
University of Chicago
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
005421136
City
Chicago
State
IL
Country
United States
Zip Code
60637
Girard, Romuald; Khanna, Omaditya; Shenkar, Robert et al. (2016) Peripheral plasma vitamin D and non-HDL cholesterol reflect the severity of cerebral cavernous malformation disease. Biomark Med 10:255-64
Tabit, Corey E; Chen, Phetcharat; Kim, Gene H et al. (2016) Elevated Angiopoietin-2 Level in Patients With Continuous-Flow Left Ventricular Assist Devices Leads to Altered Angiogenesis and Is Associated With Higher Nonsurgical Bleeding. Circulation 134:141-52
Hofmann Bowman, Marion A; Liao, James K (2016) Relative Lack of Culprit and Obstructive Coronary Lesions in Patients With Acute Ischemic Stroke and Elevated Cardiac Troponin. Circulation 133:1228-9
Lennon, Frances E; Cianci, Gianguido C; Kanteti, Rajani et al. (2016) Unique fractal evaluation and therapeutic implications of mitochondrial morphology in malignant mesothelioma. Sci Rep 6:24578
Kasahara, David I; Mathews, Joel A; Ninin, Fernanda M C et al. (2016) Role of ROCK2 in CD4(+) cells in allergic airways responses in mice. Clin Exp Allergy :
Kajikawa, Masato; Noma, Kensuke; Nakashima, Ayumu et al. (2015) Rho-associated kinase activity is an independent predictor of cardiovascular events in acute coronary syndrome. Hypertension 66:892-9
Knipe, Rachel S; Tager, Andrew M; Liao, James K (2015) The Rho kinases: critical mediators of multiple profibrotic processes and rational targets for new therapies for pulmonary fibrosis. Pharmacol Rev 67:103-17
Hsieh, Min-Ling; Liu, Ping-Yen; Wu, Jing-Ming et al. (2015) Interventional Transcatheter Closure Ameliorates the Leukocyte Rho Kinase Activities among Patients with Patent Ductus Arteriosus. Acta Cardiol Sin 31:494-9
Kasahara, David I; Ninin, Fernanda M C; Wurmbrand, Alison P et al. (2015) Abrogation of airway hyperresponsiveness but not inflammation by rho kinase insufficiency. Clin Exp Allergy 45:457-70
Liao, Yi-Chu; Liu, Ping-Yen; Lin, Hsiu-Fen et al. (2015) Two functional polymorphisms of ROCK2 enhance arterial stiffening through inhibiting its activity and expression. J Mol Cell Cardiol 79:180-6

Showing the most recent 10 out of 137 publications