B-arrestin Biased B1- and B2-Adrenergic Receptor Signaling

B-arrestin 偏向 B1- 和 B2- 肾上腺素能受体信号转导

基本信息

批准号：
8382594
负责人：
Howard A Rockman
金额：
$ 35.36万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8382594
关键词：
ADRB1 gene ADRB2 gene Adenoviruses Adopted Adrenergic Receptor Adult Agonist Apoptosis Arrestins Biological Assay Bioluminescence Biosensor Cardiac Cardiac Myocytes Catecholamines Cell Survival Chronic Data Development Energy Transfer Engineering G-Protein-Coupled Receptors GTP-Binding Proteins Gene Targeting Heart Heart failure Human Ischemia Knock-in Mouse Lead Ligands Mediating Metabolic Molecular Molecular Conformation Mus Muscle Cells Myocardial Infarction Oryctolagus cuniculus Pathway interactions Phenotype Phosphoproteins Physiological Poly(ADP-ribose) Polymerases Proteins Proteomics Receptor Signaling Recruitment Activity Reperfusion Therapy Research Personnel Role Signal Pathway Signal Transduction Stimulus Stress Testing Therapeutic Agents Virus Work abstracting arrestin B arrestin3 desensitization estin in vivo mutant novel pressure protein activation receptor response trafficking

项目摘要

Abstract B-arrestins are multifunctional proteins that are recruited to G protein-coupled receptors (GCPRs) following agonist stimulation. While the classical role of (3-arrestin is to mediate receptor desensitization, work by investigators of this PPG have recently shown that P-arrestin can stimulate signaling in the absence of classical G protein activation. The existence of B-arrestin-mediated signaling independent of G proteins requires that receptors adopt multiple "active" conformations or "ligand selective states". The ability of unique ligand-receptor conformations to promote preferential B-arrestin signaling is an emerging concept known as "biased signaling". The molecular mechanisms that underlie p-an-estin-biased signaling for the p-adrenergic receptor of (PAR), and its physiological consequences in the heart, are not known. In this proposal, we will test the hypothesis that mutant p i - and P2 can be engineered that will selectively stimulate p-arrestinbiased signaling independent of G protein activation, and that p-arrestin-biased signaling will promote cardiomyocyte cell survival to limit the development of heart failure in response to pathological stimuli. Accordingly, the specific aims of the study are: Aim 1: To engineerBp1 AR mutants that show selective bias for p-arrestin recruitment. Aim 2: To identify the mechanism of activation and signaling pathways activated by P1AR and B2AR mutants in the absence of G protein activation. Aim 3: To test in adult cardiomyocytes whether p-arrestin-biasedBP2AR TYY and B1 AR mutants activate cardioprotective signaling in response to agonist stimulation and ischemia. Aim 4: To test in vivo whether the B-arrestin-biased Bp2AR TYY and pi AR mutant activities cardioprotective pathways under conditions of pathological stress. By exploring these aims, we will define the pathways by which G protein-Independent activation of BARs may lead to stimulation of cardioprotective signaling. If our hypothesis is correct, we will show that ligandstimulated PARS, which selectively activate B-arrestin signaling pathways, are cardioprotecitve. Since, by definition, the administration of a ligand that does not stimulate G protein signaling is B-blackade, we will have demonstrated proof-of concept for the development of an entirely novel class of receptor blockers. We believe these data will provide considerable impetus for the development of novel p-arrestin-biased therapeutic agents to treat human heart failure.

抽象的 B-抑制蛋白是多功能蛋白，在激动剂刺激后被招募到 G 蛋白偶联受体 (GCPR)。虽然 (3-arrestin) 的经典作用是介导受体脱敏，但该 PPG 研究人员最近的工作表明，P-arrestin 可以在缺乏经典的 G 蛋白激活。 B-抑制蛋白介导的信号传导独立于 G 蛋白的存在，要求受体采用多种“活性”构象或“配体选择性状态”。独特的能力促进优先 B-抑制蛋白信号传导的配体-受体构象是一个新兴概念，称为“偏向信号传导”。 p-肾上腺素能偏向 p-an-estin 信号传导的分子机制 (PAR) 受体及其在心脏中的生理影响尚不清楚。在本提案中，我们将测试这样的假设：突变体 p i - 和 P2 可以被设计为选择性地刺激 p-arrestin 偏向信号传导，而与 G 蛋白激活无关，并且 p-arrestin 偏向信号传导将促进心肌细胞存活，从而限制心肌细胞的发展。对病理刺激的反应导致心力衰竭。因此，本研究的具体目标是：目标 1：设计 Bp1 AR 突变体，使其对 p-arrestin 募集表现出选择性偏差。目标 2：确定激活机制以及在没有 G 蛋白激活的情况下 P1AR 和 B2AR 突变体激活的信号通路。目标 3：在成人心肌细胞中测试 p-arrestin 偏向的 BP2AR TYY 和 B1 AR 突变体是否会响应激动剂刺激和缺血而激活心脏保护信号。目标 4：体内测试偏向 B-arrestin 的 Bp2AR TYY 和 pi AR 突变体在病理应激条件下是否具有心脏保护途径。通过探索这些目标，我们将定义 BAR 的 G 蛋白独立激活可能导致心脏保护信号传导刺激的途径。如果我们的假设是正确的，我们将证明配体刺激 PARS 可选择性激活 B-arrestin 信号通路，具有心脏保护作用。由于根据定义，施用不刺激 G 蛋白信号传导的配体是 B-blackade，因此我们将证明开发一类全新的受体阻滞剂的概念验证。我们相信这些数据将为开发新型 p-抑制蛋白偏向治疗剂来治疗人类心力衰竭提供相当大的推动力。