Regulation of Vascular Inflammatory Signaling by the Deubiquitinase USP20

去泛素酶 USP20 对血管炎症信号的调节

基本信息

批准号：
9893026
负责人：
NEIL J. FREEDMAN
金额：
$ 53.46万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-03-15 至 2023-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9893026
关键词：
Adaptor Signaling Protein Adenoviruses Amino Acids Anti-Inflammatory Agents Antiatherogenic Arrestins Arterial Injury Atherosclerosis Attenuated Binding Biological Assay Blood Vessels CCL2 gene CRISPR/Cas technology Cause of Death Cell Adhesion Molecules Chronic Cytokine Receptors Deubiquitination Dominant-Negative Mutation Endothelial Cells Endothelium Family Goals Human Hyperplasia I Kappa B-Alpha IRAK1 gene In Vitro Industrialization Inflammation Inflammatory Interleukin 4 Receptor Interleukin-1 Receptors LOX gene Link Lipopolysaccharides Mediating Molecular Mus Myocardial Infarction Partner in relationship Phosphorylation Phosphotransferases Polyubiquitination Post-Translational Protein Processing Protein Kinase Proteins RIPK1 gene ROCK1 gene Regulation Role Signal Pathway Signal Transduction Signaling Protein Smooth Muscle Myocytes Stroke TLR2 gene TLR4 gene TNF gene TNFRSF1A gene TRAF2 gene TRAF6 gene Tamoxifen Testing Transgenic Mice Transgenic Organisms Tumor Necrosis Factor Receptor Ubiquitin Ubiquitination atherogenesis cellular transduction congenic cytokine desensitization in vivo interleukin-1 receptor-associated kinase loss of function lysophosphatidic acid macrophage mutant new therapeutic target novel protein B response scaffold transcription factor ubiquitin-protein ligase ubiquitin-specific protease vascular inflammation western diet

项目摘要

PROJECT SUMMARY We recently demonstrated that ubiquitin-specific protease-20 (USP20) is scaffolded by the adaptor protein known as β-arrestin2 (βarr2), and that USP20 desensitizes ubiquitin-dependent signaling from Toll-like receptor-4 (TLR4) to NFκB activation by deubiquitinating TRAF6 and βarr2. Using transgenic mice expressing dominant-negative USP20 in smooth muscle cells, we found that USP20 reduces neointimal hyperplasia after arterial injury and that USP20 activity in SMCs reduces atherosclerosis in Ldlr-/- mice. To establish anti-atherogenic effects of systemically expressed USP20, and to elucidate further molecular mechanisms by which USP20 protects against atherosclerosis, this project will test the hypothesis that USP20 attenuates atherosclerosis by deubiquitinating several substrate proteins that were previously unassociated with USP20 but that are important in signaling pathways that activate NFκB: βarr1, TRAF6, TRAF2, and RIPK1. Furthermore, because USP20 employs βarr2 as a scaffold to facilitate association with distinct proteins, and because βarr1 reduces vascular inflammation, this project will test whether USP20’s anti-atherogenic activity involves βarr1-mediated scaffolding. To these ends, this project will study systemic effects of USP20 on atherosclerosis by comparing Usp20-/- /Ldlr-/- versus Ldlr-/- mice, on a background of βarr1+/+ or βarr1-/+. To determine the effects of endothelial USP20 on atherosclerosis, this project will compare atherosclerosis among VECad-Cre- ERT2/Usp20flox/flox/Ldlr-/- vs. Usp20flox/flox/Ldlr-/- mice treated ± tamoxifen; furthermore, we will investigate cytokine secretion, and dynamic ubiquitination of signaling proteins in primary aortic endothelial cells that are WT, Usp20-/-, Usp20-/-/βarr1-/+, or βarr1-/-. To determine what kinase in endothelial cells phosphorylates USP20 on Ser333 (and thereby abrogates USP20 deubiquitinase activity), this project will test IRAK1, PAK1, and ROCK1 with several loss-of function approaches, including a USP20 minigene, in primary endothelial cells. These studies collectively may identify USP20 phosphorylation as novel therapeutic target for atherosclerosis.

项目摘要我们最近证明了泛素特异性蛋白酶-20（USP20）由适配器脚手架蛋白质称为β-arrestin2（βarr2），USP20脱敏泛素依赖性信号传导通过去泛素化TRAF6和βARR2，Toll样受体-4（TLR4）至NFκB激活。使用转基因在平滑肌细胞中表达显性阴性USP20的小鼠，我们发现USP20降低伪影后的新内膜增生和SMC中的USP20活性降低了动脉粥样硬化 LDLR - / - 小鼠。建立系统表达的USP20的抗动脉生就性，并阐明 USP20预防动脉粥样硬化的进一步的分子机制，该项目将测试 USP20通过去泛素化几种底物蛋白来减轻动脉粥样硬化的假设以前与USP20无关，但在激活的信号通路中很重要 NFκB：βARR1，TRAF6，TRAF2和RIPK1。此外，因为USP20员工βarr2作为脚手架为了促进与不同蛋白质的关联，并且由于βARR1降低了血管感染，这项目将测试USP20的抗动脉粥样硬化活性是否涉及βarr1介导的脚手架。到这些目的是，该项目将通过比较USP20 - / - 研究USP20对动脉粥样硬化的系统影响 /ldlr - / - 与ldlr - / - 小鼠，在βarr1+/+或βarr1-/+的背景下。确定内皮的影响 USP20在动脉粥样硬化方面，该项目将比较Vecad-Cre-中的动脉粥样硬化 ERT2/USP20FLOX/FLOX/LDLR - / - vs. USP20FLOX/FLOX/LDLR - / - 小鼠接受治疗±tamoxifen;此外，我们将调查一级主动脉内皮细胞中信号蛋白的细胞因子分泌和动态泛素化 wt，USP20 - / - ，USP20 - / - /βarr1 - /+或βarr1 - / - 。确定内皮细胞中的激酶磷酸化Ser333上的USP20（从而消除了USP20去泛素酶活性），该项目将使用多种功能丧失方法测试IRAK1，PAK1和ROCK1，包括USP20 在原发性内皮细胞中的微基因。这些研究可以集体鉴定USP20磷酸化作为动脉粥样硬化的新型治疗靶标。