Studies in this laboratory have elucidated the role of redox regulation of signaling in vascular cell function. Amongst other redox-sensitive proteins, we found that S-glutathiolation of p21ras cysteine-118 (C118) by reactive nitrogen species (RNS), including 7NO and low concentrations of peroxynitrite (OONO-), acutely increases its GTPase activity and MAP- and PI3-kinase signaling. Migration of endothelial cells (EC) which is required for angiogenesis during tissue repair is also mediated by 7NO, but the redox-regulated protein targets of 7NO that are important for mediating angiogenesis are unknown. Our preliminary studies show that expression of wild type (WT) p21ras, but not a C118 serine (C118S) mutant induces EC migration. Furthermore, overexpression of glutaredoxin-1 (GRX1), inhibits EC migration stimulated by VEGF, 7NO, and WT p21ras, suggesting that S-glutathiolation of the p21ras cysteine-118 thiol is essential. The proposed studies will therefore investigate the redox regulation of p21ras that supports normal EC migration which is stimulated by 7NO and HMG CoA reductase inhibitors (statins). Our preliminary studies suggest that statins, by interfering with farnesylation of the C-terminal cysteine which normally keeps p21ras tethered to the plasma membrane, facilitate relocation of p21ras to mitochondria, thereby increasing mitochondrial fission, ROS/RNS production, and p21ras-mediated, redox-dependent angiogenic signaling. We will also investigate whether exposure of EC to oxidants and reactive lipids associated with inflammation and metabolic vascular disease disrupts normal p21ras activity and/or subcellular localization. Results of these studies in EC in culture will be relevant for our studies in obese mice fed a high fat, high sucrose (HFHS) diet in which we find angiogenesis to be impaired. Our preliminary studies also indicate that angiogenesis is diminished in the ischemic hindlimb of GRX1 transgenic mice, suggesting the hypothesis that altered GRX1 expression in metabolic disease may inhibit p21ras and angiogenesis. By understanding the molecular mechanisms by which angiogenesis is normally redox-regulated by p21ras and GRX1, we anticipate being able to determine why angiogenesis is impaired in metabolic disease.

Public Health Relevance

Cardiac and limb ischemia caused by diabetic vascular disease is due in part to impaired angiogenesis, the formation of new blood vessels. We propose that angiogenesis is impaired because oxidants and lipids prevent the normal chemical reactions between nitric oxide and the signaling protein, p21ras which controls angiogenesis. Our studies will not only help explain why angiogenesis is impaired in diabetes, but also provide insight into the therapeutic mechanism of HMG CoA reductase inhibitors (statins) and novel strategies to improve diabetic vascular disease.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37HL104017-03
Application #
8294637
Study Section
Atherosclerosis and Inflammation of the Cardiovascular System Study Section (AICS)
Program Officer
Gao, Yunling
Project Start
2010-07-15
Project End
2015-06-30
Budget Start
2012-07-01
Budget End
2013-06-30
Support Year
3
Fiscal Year
2012
Total Cost
$442,408
Indirect Cost
$170,157
Name
Boston University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
604483045
City
Boston
State
MA
Country
United States
Zip Code
02118
Watanabe, Yosuke; Murdoch, Colin E; Sano, Soichi et al. (2016) Glutathione adducts induced by ischemia and deletion of glutaredoxin-1 stabilize HIF-1α and improve limb revascularization. Proc Natl Acad Sci U S A 113:6011-6
Peskin, Alexander V; Pace, Paul E; Behring, Jessica B et al. (2016) Glutathionylation of the Active Site Cysteines of Peroxiredoxin 2 and Recycling by Glutaredoxin. J Biol Chem 291:3053-62
Tong, Xiaoyong; Khandelwal, Alok R; Wu, Xiaojuan et al. (2016) Pro-atherogenic role of smooth muscle Nox4-based NADPH oxidase. J Mol Cell Cardiol 92:30-40
Cohen, Richard A; Murdoch, Colin E; Watanabe, Yosuke et al. (2016) Endothelial Cell Redox Regulation of Ischemic Angiogenesis. J Cardiovasc Pharmacol 67:458-64
Ji, Yuhuan; Bachschmid, Markus M; Costello, Catherine E et al. (2016) S- to N-Palmitoyl Transfer During Proteomic Sample Preparation. J Am Soc Mass Spectrom 27:677-85
Yao, Chunxiang; Behring, Jessica B; Shao, Di et al. (2015) Overexpression of Catalase Diminishes Oxidative Cysteine Modifications of Cardiac Proteins. PLoS One 10:e0144025
Tong, Xiaoyong; Khandelwal, Alok R; Qin, Zhexue et al. (2015) Role of smooth muscle Nox4-based NADPH oxidase in neointimal hyperplasia. J Mol Cell Cardiol 89:185-94
Mei, Yu; Thompson, Melissa D; Cohen, Richard A et al. (2015) Autophagy and oxidative stress in cardiovascular diseases. Biochim Biophys Acta 1852:243-51
Seta, Francesca; Cohen, Richard A (2014) The endothelium: paracrine mediator of aortic dissection. Circulation 129:2629-32
Mei, Yu; Thompson, Melissa D; Shiraishi, Yasunaga et al. (2014) Sarcoplasmic/endoplasmic reticulum Ca2+ ATPase C674 promotes ischemia- and hypoxia-induced angiogenesis via coordinated endothelial cell and macrophage function. J Mol Cell Cardiol 76:275-82

Showing the most recent 10 out of 32 publications