Mechanism and Function of Beta-2 Adrenergic Receptor Degradation in the Lung

肺内β2肾上腺素受体降解的机制和功能

基本信息

批准号：
8701380
负责人：
Quan Lu
金额：
$ 40.62万
依托单位：
HARVARD SCHOOL OF PUBLIC HEALTH
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-07-15 至 2017-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8701380
关键词：
Acute Adenylate Cyclase Adrenergic Receptor Affect Agonist Albuterol Animal Model Arrestins Asthma Biology Bronchodilator Agents Cell model Cells Chronic Chronic Obstructive Airway Disease Chronic lung disease Clinical Clinical Trials Cyclic AMP Data Degradation Pathway Development Disease Down-Regulation Genes Genetic Genetic Variation Genotype House Dust Mite Allergens Human Human Biology In Vitro Individual Inflammation Inflammatory Lead Link Lung Measures Mediating Molecular Molecular and Cellular Biology Mus Muscle function Muscle relaxation phase Pathway interactions Patients Pharmacogenetics Phenotype Physiological Physiology Play Population Price Process Production Proteins RNA Interference Recruitment Activity Regulation Role Signal Transduction Smooth Muscle Myocytes Stimulus Testing Therapeutic Ubiquitination Variant Work airway inflammation allergic airway disease arrestin3 base beta-2 Adrenergic Receptors cytokine density functional outcomes gene function genome wide association study genome-wide improved in vivo novel prevent public health relevance receptor receptor function research study respiratory smooth muscle response ubiquitin-protein ligase

项目摘要

DESCRIPTION (provided by applicant): The ?2-adrenergic receptor (?2AR) is central in normal lung physiology and disease, but key questions about its regulation remain unanswered. Since ?2AR signaling causes airway smooth muscle relaxation, agonists of the receptor are a mainstay therapy for asthma and COPD. Receptor protein down-regulation is a critical mechanism for controlling ?2AR, and works through ubiquitination and subsequent degradation of ?2AR upon prolonged agonist stimulation. This homeostatic mechanism prevents potentially detrimental persistent activation but comes with a price: the efficacy of ?-agonist therapy is often reduced by the very same process. A better understanding of ?2AR degradation would thus benefit both basic receptor biology and translational ?-agonist therapy. Using a unique genome- wide RNA interference (RNAi) screen, we discovered ?-arrestin3, which is encoded by a novel human gene ARRDC3 (arrestin domain containing 3), as a critical new component of the ?2AR degradation machinery. Alpha-arrestin-3 co-localizes and interacts with ?2AR in an agonist-dependent manner. Through a specific interaction, ?-arrestin3 recruits the ubiquitin E3 ligase NEDD4-1 to activated ?2AR to mediate the receptor ubiquitination and subsequent degradation. Building upon these molecular studies, we now propose to examine the function of ?-arrestin3-mediated ?2AR regulation pathway at the physiologic and human biology levels. We hypothesize that ?-arrestin3-mediated ?2AR degradation plays an important physiological role in the lung and that perturbation of the degradation pathway impacts all aspects of functional outcomes of ?2AR signaling. The proposal is to test this hypothesis with three Aims. In Aim 1 we will determine the role of ?-arrestin3 in regulating the physiological function of ?2AR in airway smooth cells. Aim 2 will test the role of ?-arrestin3 in regulating the bronchoprotection provided by ¿-agonist administration in mice in vivo. Aim 3 will investigate the association of genetic variations in the ?-arrestin3-mediated ?2AR degradation pathway with bronchodilator response (BDR) in asthma patients. This highly integrative project combines molecular and cellular biology, animal model, and pharmacogenetics and will establish a critical role for ?-arrestin3 in regulating ?2AR functions in the lung. Moreover, the results will lead to a better understanding of the genetic basis of individual ?-agonist responses, and may ultimately contribute to the development of improved and personalized ?-agonist therapy for asthma and other chronic lung diseases.

描述（由申请人提供）：β2-肾上腺素能受体（β2AR）在正常肺生理学和疾病中至关重要，但有关其调节的关键问题仍未得到解答，因为β2AR信号传导导致气道平滑肌松弛，因此该受体的激动剂是重要的。哮喘和慢性阻塞性肺病的主要治疗方法受体蛋白下调是控制 ?2AR 的关键机制，并通过 ?2AR 长时间泛素化和随后的降解发挥作用。这种稳态机制可以防止潜在的痛苦的持续激活，但也有代价：β-激动剂治疗的功效通常会因相同的过程而降低，因此更好地了解β2AR降解将有利于基础受体生物学和转化β。使用独特的全基因组 RNA 干扰 (RNAi) 筛选，我们发现了 β-arrestin3，它由新型人类基因 ARRDC3（包含 3 的抑制蛋白结构域）编码，是一种重要的新成分。 α-arrestin-3 以激动剂依赖性方式与 β2AR 共定位并相互作用，通过特定的相互作用，α-arrestin3 招募泛素 E3 连接酶 NEDD4-1 来激活 β2AR。基于这些分子研究，我们现在建议检查 β-arrestin3 介导的 β2AR 调节途径的功能。我们勇敢地承认，β-arrestin3 介导的 β2AR 降解在肺中起着重要的生理作用，并且降解途径的扰动会影响 β2AR 信号传导功能的各个方面。具有三个目标。在目标 1 中，我们将确定 β-arrestin3 在调节气道平滑细胞中 β2AR 的生理功能中的作用。目标 2 将测试 β-arrestin3 在调节气道平滑细胞中的作用。调节 ¿ 提供的支气管保护目标 3 将研究 β-arrestin3 介导的 β2AR 降解途径中的遗传变异与哮喘患者的支气管扩张反应 (BDR) 之间的关系。和药物遗传学，并将确定 β-arrestin3 在调节肺中 β2AR 功能中的关键作用。更好地了解个体β-激动剂反应的遗传基础，并可能最终有助于开发针对哮喘和其他慢性肺部疾病的改进和个性化β-激动剂疗法。