Arrestin Selectivity for GPCRs in Airway Smooth Muscle

Arrestin 对气道平滑肌中 GPCR 的选择性

• 批准号: 8461974
• 负责人: RAYMOND B. PENN
• 金额: $ 37.68万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2014-10-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8461974
• 关键词: ARRB2; Ablation; Acetylcholine; Acute; Affect; Agonist; Airway Resistance; Allergic inflammation; Arrestin Beta 1; Arrestins; Asthma; Binding; Biological Assay; Biological Models; Breathing; Bronchoconstriction; Calcium; Cell Line; Cells; Chronic; Chronic Obstructive Airway Disease; Contracts; Data; Degradation Pathway; Dinoprostone; Disease; Equilibrium; Event; Exposure to; G Protein-Coupled Receptor Signaling; G alpha q Protein; G protein coupled receptor kinase; G-Protein-Coupled Receptors; GTP-Binding Proteins; Genes; Goals; Histamine; Histamine Receptor; Human; In Vitro; Inflammatory; Link; Measurement; Measures; Mediating; Modeling; Molecular Biology; Molecular Genetics; Mus; Muscarinic Antagonists; Muscarinics; Muscle Contraction; Ovalbumin; Pharmaceutical Preparations; Phospholipase C; Physiological; Process; Property; Protein Isoforms; Protocols documentation; Recombinants; Recycling; Regulation; Relative (related person); Relaxation; Role; Signal Transduction; Smooth Muscle; Smooth Muscle Myocytes; Steroids; Stimulus; System; Testing; Therapeutic; Time; Tissues; Transducers; Treatment Efficacy; Wheezing; airway hyperresponsiveness; attenuation; base; beta-2 Adrenergic Receptors; cell type; constriction; cysteinyl leukotriene receptor; cysteinyl-leukotriene; desensitization; improved; in vivo; overexpression; prevent; public health relevance; receptor; receptor coupling; receptor internalization; receptor-mediated signaling; release of sequestered calcium ion into cytoplasm; respiratory smooth muscle; response; trafficking

DESCRIPTION (provided by applicant): Arrestins regulate numerous G protein-coupled receptors (GPCRs) by disrupting GPCR-G protein interaction and "arresting" the ability of receptors to signal. Although considerable evidence of the importance of arrestins has come from studies employing cellular overexpression systems, our understanding of arrestin regulation of endogenous GPCRs in physiological systems is limited. We propose to characterize the capacity and selectivity of arrestins for regulating GPCRs in airway smooth muscle (ASM), with the ultimate goal of targeting arrestins as a means of improving the therapeutic efficacy of inhaled beta-agonists in the treatment of asthma and COPD. Based on preliminary data presented herein, we hypothesize that differences in beta-arrestin-1 and beta-arrestin-2 selectivity for Gq- and Gs- coupled receptors can be exploited to enhance the ability of beta-agonists to relax ASM contracted by other GPCRs. Aims 1 & 2 will establish the selectivity of beta-arrestin-1 and beta-arrestin-2 for those GPCRs that either mediate contraction (including muscarinic, cysteinyl leukotriene, and histamine receptors) or relaxation (beta-2-adreneric and prostaglandin E2 receptors) of ASM. Aim 1 will utilize molecular and genetic approaches to target beta-arrestin-1, beta-arrestin-2, or both in ASM cells and tissue derived from both humans and mice. Affects of arrestin inhibition or gene ablation on both ASM signaling and contractile properties by GPCRs will be assessed. Aim 2 will perform complementary physiological measurements to determine the effect of arrestin gene ablation on the regulation of ASM contraction and relaxation in vivo. Collectively, these studies will provide a highly mechanistic analysis of arrestin selectivity in ASM and identify approaches for differentially regulating receptors that mediate contraction or relaxation of ASM. PUBLIC HEALTH RELEVANCE: These studies seek to identify the usefulness (as an asthma therapy) of inhibiting a key regulatory molecule in airway smooth muscle. Understanding how beta-arrestin-1 or beta-arrestin-2 regulate receptors in smooth muscle may allow us to improve the ability of drugs to relax smooth muscle while at the same time inhibiting the stimuli that cause airway constriction and wheezing.

描述（由申请人提供）：逮捕蛋白通过破坏GPCR-G蛋白相互作用并“阻止”受体信号的能力来调节众多G蛋白偶联受体（GPCR）。尽管逮捕蛋白重要性的大量证据来自采用细胞过表达系统的研究，但我们对生理系统中内源性GPCR的抑制蛋白调节的理解是有限的。我们建议表征逮捕蛋白在调节气道平滑肌（ASM）中的能力和选择性，其最终目的是将逮捕蛋白靶向，以此作为提高哮喘和COPD治疗中吸入β-启动器的治疗功效的一种手段。基于本文提供的初步数据，我们假设可以利用GQ-和GS-耦合受体的β-arrestin-1和Beta-arrestin-2选择性的差异，以增强β-激动剂放宽其他GPCR合同的ASM的能力。 AIMS 1和2将建立β-arrestin-1和Beta-arrestin-2的选择性，以介导ASM的那些介导收缩（包括毒蕈碱，西胱烷基白细胞和组胺受体）的GPCR（包括毒蕈碱，西藻酰基白细胞受体和组胺受体）。 AIM 1将利用分子和遗传学方法来靶向β-arrestin-1，beta-arrestin-2，或在人类和小鼠衍生的ASM细胞和组织中。将评估对GPCR的ASM信号传导和收缩性质的抑制素抑制或基因消融的影响。 AIM 2将执行互补的生理测量，以确定抑制蛋白基因消融对体内ASM收缩和放松调节的影响。总的来说，这些研究将对ASM中的抑制蛋白选择性提供高度机械的分析，并确定调节介导ASM收缩或放松的受体的方法。 公共卫生相关性：这些研究旨在确定抑制气道平滑肌中关键调节分子的有用性（作为哮喘治疗）。了解β-arrestin-1或β-arrestin-2如何调节平滑肌中的受体可以使我们能够提高药物放松平滑肌的能力，同时抑制引起气道收缩和喘息的刺激。