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

• 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
• 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

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（通过应用提供）：2-肾上腺素能受体（？2AR）在正常的肺部生理和疾病中是中心的，但是有关其调节的关键问题仍未得到解答。由于？2AR信号导致气道平滑肌松弛，因此受体的激动剂是哮喘和COPD的主要疗法。受体蛋白下调是控制2AR的关键机制，并且在延长的激动剂刺激后通过泛素化和随后的序列降解。这种体内稳态机制可防止潜在的持续激活，但价格有所下降：激动人物治疗的效率通常会因同样的过程而降低。更好地理解？2AR降解将使基本受体生物学和转化疗法受益？激动剂治疗。使用独特的基因组宽RNA干扰（RNAi）屏幕，我们发现了？ - arrestin3，它是由新型的人类基因ARRDC3（含有3）编码的，是？2AR降解机械的关键组成部分。 alpha-arrestin-3以激动剂依赖性方式共定位并与？2AR相互作用。通过特定的相互作用，？ - arrestin3募集泛素E3连接酶Nedd4-1激活？2AR以介导接收器的泛素化和随后的降解。在这些分子研究的基础上，我们现在建议检查-Arrestin3介导的？2AR调节途径在生理和人类生物学水平上的功能。我们假设这是？ - arrestin3介导的？2AR降解在肺中起重要的身体作用，并且降解途径的扰动会影响？2AR信号传导功能结果的所有方面。该提议是用三个目标检验这一假设。在AIM 1中，我们将确定-Arrestin3在调节气道平滑细胞中的生理功能中的作用。 AIM 2将测试？ - arrestin3在调节体内小鼠中的支气管保护作用中的作用。 AIM 3将研究哮喘患者的遗传变异与2AR介导的？2AR降解途径与支气管扩张剂反应（BDR）的关联。这个高度整合的项目结合了分子和细胞生物学，动物模型和药物遗传学，并将在调节肺中调节？2AR功能中建立关键作用。而且，结果将导致 更好地理解个体的遗传基础？激动人心的反应，最终可能有助于改善和个性化的哮喘和其他慢性肺部疾病的激动人物治疗。