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（由适用提供）：动脉粥样硬化仍然是美国的主要死亡原因。积累的证据表明，主动脉壁平滑肌细胞基因表达对动脉粥样硬化有很大贡献。 ß-arrestin2（ßarr2）G蛋白偶联受体（GPCR），生长因子受体和离子通道的内吞和信号衔接子也已知小鼠的新内膜增生和动脉粥样硬化。由去泛素酶（DUBS）调节的可逆ßarr2泛素化是一种关键的翻译后修饰，是ßarr2的适配器在介导细胞信号时所必需的。我们的初步研究表明，DUB USP20可能会影响ßarr2泛素化以及由动脉粥样硬化Toll样受体4（TLR4）诱导的NFκB信号传导。要描述体内泛泛素化/去泛素化动力学的作用，并评估ßarr2的泛素化状态是否可以在SMC中引起促发炎信号，我们已经在USP20产生了转基因小鼠，表达了USP20或表达其催化性的SPERMOFERMEFERM（DN-SPEC22）。在该模型中，我们预计，通过去泛素化β2，USP20将减少ßarr2活性，从而减少SMC亲动脉粥样硬化的增殖和迁移，从而导致新的增生增生，而DN-USP20会产生相互影响的效果。 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通过实现特定目的进行脚手架：（1）确定SMC USP20的动脉保护作用，以及USP20的作用机理是否需要对ßarr2（2）的泛素化进行去泛素化，以调节USP20活性确定eplym2依赖性SMC的迁移和信号的效果是否触发了（3）的机构（3） ßarr2依赖性信号传导。