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：
8699256
负责人：
RICHARD A COHEN
金额：
$ 43.36万
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/8699256
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 adenoviral-mediated angiogenesis cell motility chemical reaction cytokine farnesylation feeding improved insight matrigel migration mutant novel strategies overexpression prevent response subcutaneous tissue repair

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.

描述（由申请人提供）：本实验室的研究阐明了信号传导的氧化还原调节在血管细胞功能中的作用。在其他氧化还原敏感蛋白中，我们发现 p21ras 半胱氨酸 118 (C118) 通过活性氮 (RNS)（包括 7NO 和低浓度过氧亚硝酸盐 (OONO-)）进行 S-谷胱甘肽化，急剧增加其 GTP 酶活性和 MAP- 和PI3 激酶信号传导。组织修复过程中血管生成所需的内皮细胞 (EC) 的迁移也是由 7NO 介导的，但对于介导血管生成很重要的 7NO 氧化还原调节蛋白靶标尚不清楚。我们的初步研究表明，野生型 (WT) p21ras 的表达（而非 C118 丝氨酸 (C118S) 突变体）会诱导 EC 迁移。此外，谷氧还蛋白-1 (GRX1) 的过表达会抑制 VEGF、7NO 和 WT p21ras 刺激的 EC 迁移，表明 p21ras 半胱氨酸 118 硫醇的 S-谷胱甘肽化是必需的。因此，拟议的研究将调查 p21ras 的氧化还原调节，该调节支持由 7NO 和 HMG CoA 还原酶抑制剂（他汀类药物）刺激的正常 EC 迁移。我们的初步研究表明，他汀类药物通过干扰 C 端半胱氨酸的法呢基化（通常将 p21ras 束缚在质膜上），促进 p21ras 重新定位到线粒体，从而增加线粒体裂变、ROS/RNS 产生以及 p21ras 介导的氧化还原依赖性血管生成信号传导。我们还将研究 EC 暴露于与炎症和代谢性血管疾病相关的氧化剂和反应性脂质是否会破坏正常的 p21ras 活性和/或亚细胞定位。这些培养 EC 的研究结果将与我们对喂食高脂肪、高蔗糖 (HFHS) 饮食的肥胖小鼠的研究相关，我们发现在这些小鼠中血管生成受到损害。我们的初步研究还表明，GRX1 转基因小鼠缺血后肢的血管生成减少，这表明代谢疾病中 GRX1 表达改变可能抑制 p21ras 和血管生成的假设。通过了解血管生成通常由 p21ras 和 GRX1 氧化还原调节的分子机制，我们预计能够确定代谢疾病中血管生成受损的原因。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Both hydrogen peroxide and transforming growth factor beta 1 contribute to endothelial Nox4 mediated angiogenesis in endothelial Nox4 transgenic mouse lines.

在内皮 Nox4 转基因小鼠系中，过氧化氢和转化生长因子 β 1 均有助于内皮 Nox4 介导的血管生成。

DOI：
10.1016/j.bbadis.2014.10.007
发表时间：
2014-12-01
期刊：
Biochimica et biophysica acta
影响因子：
0
作者：
Lili Chen;Jennifer Xiao;J. Kuroda;T. Ago;J. Sadoshima;R. Cohen;Xiaoyong Tong
通讯作者：
Xiaoyong Tong

Oxidative inhibition of the vascular Na+-K+ pump via NADPH oxidase-dependent β1-subunit glutathionylation: implications for angiotensin II-induced vascular dysfunction.

通过 NADPH 氧化酶依赖性 β1 亚基谷胱甘肽氧化抑制血管 Na -K 泵：对血管紧张素 II 诱导的血管功能障碍的影响。

DOI：
发表时间：
2013-12
期刊：
影响因子：
7.4
作者：
Liu, Chia;Karimi Galougahi, Keyvan;Weisbrod, Robert M;Hansen, Thomas;Ravaie, Ramtin;Nunez, Andrea;Liu, Yi B;Fry, Natasha;Garcia, Alvaro;Hamilton, Elisha J;Sweadner, Kathleen J;Cohen, Richard A;Figtree, Gemma A
通讯作者：
Figtree, Gemma A

Regulation of neovascularization by S-glutathionylation via the Wnt5a/sFlt-1 pathway.

S-谷胱甘肽通过 Wnt5a/sFlt-1 途径调节新血管形成。

DOI：
10.1042/bst20140213
发表时间：
2014-12
期刊：
Biochemical Society transactions
影响因子：
3.9
作者：
Murdoch CE;Bachschmid MM;Matsui R
通讯作者：
Matsui R

Glutathione peroxidase-1 deficiency potentiates dysregulatory modifications of endothelial nitric oxide synthase and vascular dysfunction in aging.

谷胱甘肽过氧化物酶-1 缺乏会加剧衰老过程中内皮一氧化氮合酶的失调修饰和血管功能障碍。

DOI：
发表时间：
2014-02
期刊：
影响因子：
0
作者：
Oelze, Matthias;Kröller;Steven, Sebastian;Lubos, Edith;Doppler, Christopher;Hausding, Michael;Tobias, Silke;Brochhausen, Christoph;Li, Huige;Torzewski, Michael;Wenzel, Philip;Bachschmid, Markus;Lackner, Karl J;Schulz, Eberhard
通讯作者：
Schulz, Eberhard

Redox Regulation of Ischemic Angiogenesis　- Another Aspect of Reactive Oxygen Species.

缺血性血管生成的氧化还原调节——活性氧的另一个方面。