Regulation of B-arrestin2's pro-atherogenic activity by the deubiquitinase USP20

去泛素酶 USP20 对 B-arrestin2 促动脉粥样硬化活性的调节

基本信息

批准号：
8639259
负责人：
NEIL J. FREEDMAN
金额：
$ 47.46万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-02-03 至 2018-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8639259
关键词：
Affect Anti-Inflammatory Agents Anti-inflammatory Antiatherogenic Antibodies Atherosclerosis Binding Blood Vessels Breeding Cause of Death Cell Proliferation Custom Cytokine Receptors Deubiquitination Diet G-Protein-Coupled Receptors Gene Expression Genes Goals Growth Factor Receptors Health Human Hyperplasia IRAK1 gene In Vitro Inflammatory Interleukin-1 Receptors Ion Channel Ions Link MAPK3 gene Mediating Methods Modeling Molecular Mus Phosphorylation Phosphotransferases Physiological Polyubiquitination Post-Translational Protein Processing Preparation Process Protein Dephosphorylation Protein Isoforms Proteins Reagent Regulation Role Signal Transduction Signaling Protein Smooth Muscle Myocytes Stimulus System TNF gene TRAF2 gene TRAF6 gene Techniques Testing Transgenic Mice Tumor Necrosis Factor Receptor Ubiquitination United States atherogenesis atheroprotective base cell motility congenic cytokine feeding in vivo interleukin-1 receptor-associated kinase migration new therapeutic target novel prevent promoter prototype response scaffold stress-activated protein kinase 1 toll-like receptor 4 transcription factor ubiquitin-protein ligase ubiquitin-specific protease

项目摘要

DESCRIPTION (provided by applicant): Atherosclerosis remains a leading cause of death in the United States. Accumulating evidence suggests that aortic wall smooth muscle cell gene expression contributes substantially to atherogenesis. ß-arrestin2 (ßarr2) an endocytic and signaling adaptor for G protein-coupled receptors (GPCRs), growth factor receptors and ion- channels is also known to promote neointimal hyperplasia and atherosclerosis in mice. Reversible ßarr2 ubiquitination, as regulated by deubiquitinases (DUBs) is a critical post-translational modification that is required for ßarr2's adaptor functions in mediating cell-signaling. Our Preliminary Studies suggests that the DUB USP20 might affect ßarr2 ubiquitination as well as NFκB signaling induced by the atherogenic Toll-like receptor 4 (TLR4). To delineate the role(s) of ßarr2 ubiquitination/deubiquitination dynamics in vivo and to evaluate whether the ubiquitination status of ßarr2 could engender pro-inflammatory signaling in SMCs, we have generated transgenic mice expressing USP20 or its catalytically inactive isoform (DN-USP20) under control of the SMC-specific SM22α promoter. In this model, we expect that by de-ubiquitinating ßarr2, USP20 would reduce ßarr2 activity and thereby reduce the SMC pro-atherogenic proliferation and migration that engenders neointimal hyperplasia, whereas the DN-USP20 would have reciprocal effects. By utilizing these and additional novel reagents and in vivo methods involving diet and gene-dependent atherosclerosis and in vitro techniques employing primary vascular smooth muscle cells we will test the hypotheses that USP20 in SMCs mitigates atherosclerosis through mechanisms involving deubiquitination of ßarr2, and/or deubiquitination of TRAF6 or TRAF2 in a manner dependent upon ßarr2 scaffolding by accomplishing following specific aims: (1) To determine the atheroprotective role of SMC USP20, and whether USP20's mechanism of action requires de-ubiquitination of ßarr2 (2) To determine whether USP20 activity regulates ßarr2-dependent SMC proliferation, migration and signaling triggered by inflammatory stimuli and (3) To elucidate the mechanistic basis of USP20's effects on ßarr2-dependent signaling.

描述（由申请人提供）：动脉粥样硬化仍然是美国死亡的主要原因，越来越多的证据表明，主动脉壁平滑肌细胞基因表达对动脉粥样硬化形成有重要影响。偶联受体（GPCR）、生长因子受体和离子通道也被认为可促进内膜增生和动脉粥样硬化。受去泛素酶 (DUB) 调节的可逆 ßarr2 泛素化是 ßarr2 介导细胞信号转导的接头功能所必需的关键翻译后修饰。由致动脉粥样硬化 Toll 样受体 4 诱导(TLR4)。为了描述 ßarr2 泛素化/去泛素化动态在体内的作用，并评估 ßarr2 泛素化状态是否会在 SMC 中产生促炎信号传导，我们培育了表达 USP20 或其催化失活亚型的转基因小鼠 (TLR4)。 DN-USP20）在 SMC 特异性 SM22α 启动子的控制下。通过去泛素化 ßarr2，USP20 将降低 ßarr2 活性，从而减少 SMC 促动脉粥样硬化增殖和迁移，从而引起内膜增生，而 DN-USP20 通过利用这些以及其他涉及饮食的新型试剂和体内方法，会产生相反的作用。和基因依赖性动脉粥样硬化以及采用原代血管平滑肌细胞的体外技术，我们将测试 USP20 在SMC 通过涉及 ßarr2 去泛素化和/或 TRAF6 或 TRAF2 去泛素化的机制来减轻动脉粥样硬化，以依赖于 ßarr2 支架的方式实现以下具体目标： (1) 确定 SMC USP20 的动脉粥样硬化作用，以及 USP20 的作用机制是否需要ßarr2 的去泛素化 (2) 确定 USP20 活性是否调节 ßarr2 依赖性 SMC 增殖、迁移和炎症刺激触发的信号传导，以及 (3) 阐明 USP20 对 ßarr2 依赖性信号传导影响的机制基础。