Redox Regulation of p21ras in Angiogenesis

p21ras 在血管生成中的氧化还原调节

基本信息

批准号：
8109964
负责人：
RICHARD A COHEN
金额：
$ 44.24万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-15 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8109964
关键词：
1-Phosphatidylinositol 3-Kinase Accounting Blood Vessels Boston C-terminal Cardiac Cardiovascular system Cell membrane Cell physiology Collaborations Cysteine Diabetes Mellitus Diabetic Angiopathies Diet Endothelial Cells Fatty acid glycerol esters Functional disorder Grx1 protein Guanosine Triphosphate Phosphohydrolases Hindlimb Hydroxymethylglutaryl-CoA Reductase Inhibitors Inflammation Insulin Resistance Ischemia Knockout Mice Label Laboratories Limb structure Lipids Mass Spectrum Analysis Measurement Mediating Metabolic Metabolic Diseases Methods Mitochondria Mitochondrial Proteins Modification Molecular Mus Nitric Oxide Obese Mice Obesity Oxidants Oxidation-Reduction PDPK1 gene Peroxonitrite Plug-in Post-Translational Protein Processing Production Proteins Proteomics Reactive Nitrogen Species Regulation Role Serine Signal Transduction Signaling Protein Sucrose Sulfhydryl Compounds Techniques Therapeutic Transgenic Mice Universities Vascular Diseases Vascular Endothelial Growth Factors Wound Healing adenoviral-mediated angiogenesis cell motility chemical reaction cytokine farnesylation feeding improved insight matrigel migration mutant novel strategies overexpression prevent public health relevance response subcutaneous

项目摘要

DESCRIPTION (provided by applicant): 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.

描述（由申请人提供）：该实验室的研究阐明了信号传导在血管细胞功能中的作用。在其他对氧化还原敏感的蛋白中，我们发现由反应性氮种（RNS）（包括7NO和低浓度的过氧亚硝酸盐（OONO-））对P21RAS半胱氨酸-118（C118）的S-谷氨酸化（C118），急性急性增加了其GTPase活性和映射 - - 映射和图3-依3-二氧化硝酸盐（OONO-）。在组织修复过程中血管生成所必需的内皮细胞（EC）的迁移也是由7NO介导的，但是对于介导血管生成很重要的7NO的氧化还原调节的蛋白质靶标尚不清楚。我们的初步研究表明，野生型（WT）p21RA的表达，而不是C118丝氨酸（C118S）突变体诱导EC迁移。此外，谷胱甘肽-1（GRX1）的过表达抑制了VEGF，7NO和WT P21RA刺激的EC迁移，这表明P21RAS CySteine-118-118硫醇的S-谷硫化是必不可少的。因此，拟议的研究将研究支持正常EC迁移的P21RA的氧化还原调节，这是由7NO和HMG COA还原酶抑制剂（Statins）刺激的。我们的初步研究表明，他汀类药物通过干扰c-末端半胱氨酸的法尼化，通常将P21RA保持在质膜上，从而促进p21ras迁移至线粒体，从而促进线粒体散发，从而增加了线粒体的生产，ROS/RNS的产生，ROS/RNS的产生，以及P21ras-p21RIDICTIDICTIDICTIDICTIDICTIDICTIDICTIDICTIDICTIDICTIDICTIDICTIDICTIDICTIDICTIDICTIDICTIDICTIDICTIDICTIDICTIDICTIDICTIDICTIDICTIDICTIDICTIDICTIDICTIDICTICTIDICTICTIDICTICTIDID。我们还将调查EC暴露于氧化剂和与炎症和代谢血管疾病有关的反应性脂质是否会破坏正常的P21RAS活性和/或细胞下定位。培养中EC研究的这些研究的结果将与我们在肥胖小鼠中喂养高脂肪，高蔗糖（HFHS）饮食的研究有关，在这种饮食中我们发现血管生成受到损害。我们的初步研究还表明，在GRX1转基因小鼠的缺血性后肢中血管生成减少，这表明在代谢疾病中改变GRX1表达的假设可能抑制P21RAS和血管生成。通过理解通常由P21RA和GRX1氧化还原调节血管生成的分子机制，我们预计能够确定为什么在代谢疾病中损害血管生成。公共卫生相关性：由糖尿病血管疾病引起的心脏和肢体缺血部分归因于血管生成受损，即新血管的形成。我们建议血管生成受损，因为氧化剂和脂质可以防止一氧化氮和信号传导蛋白之间的正常化学反应，即控制血管生成的P21RA。我们的研究不仅会有助于解释为什么糖尿病中的血管生成受损，而且还可以洞悉HMG COA还原酶抑制剂（他汀类药物）和改善糖尿病血管疾病的新型策略。