Molecular mechanisms of TRPA1 regulation

TRPA1调节的分子机制

• 批准号: 10605343
• 负责人: Candice Elaine Paulsen
• 金额: $ 41.88万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10605343
• 关键词: Affect; Afferent Neurons; Asthma; Basic Science; Binding; Binding Sites; Biochemical; Biophysics; Calcium; Calcium Binding; Calmodulin; Cations; Chemicals; Cues; Data; Development; Disease; G-Protein-Coupled Receptors; Genetic; Human; Hyperactivity; Hypersensitivity; Inflammation; Inflammatory; Irritants; Laboratories; Mechanics; Mediating; Modeling; Molecular; Mus; Mutation; Pain; Process; Proteins; Regulation; Research; Resolution; Role; Sampling; Second Messenger Systems; Stimulus; Structure; Therapeutic; Wasabia; Work; airway inflammation; chronic pain; chronic pain management; drug development; gain of function; gain of function mutation; mutant; new therapeutic target; novel; pain chronification; pain perception; protein protein interaction; receptor; sensor; structural biology; tissue injury

PROJECT SUMMARY The wasabi receptor, TRPA1, is a non-selective homotetrameric cation channel expressed in primary sensory neurons where its activation by noxious chemical irritants contributes to pain perception and local inflammation. Local inflammatory cues, in turn, sensitize sensory neurons to painful stimuli. Within the pain and local inflammation regulatory cycle, TRPA1 serves as a positive regulator and its dysregulation could contribute to the development of chronic pain. Genetic loss of TRPA1 in mice abrogates pain perception to chemical irritants, mechanical and thermal hypersensitivity produced from tissue injury, and asthma-induced airway inflammation supporting this model. Gain-of-function TRPA1 mutations cause congenital painful disorders in humans highlighting its direct role in pain perception. This makes understanding TRPA1 function and dysregulation highly significant. Basic science research in the Paulsen Laboratory broadly aims to determine molecular mechanisms of TRPA1 regulation and dysregulation by second messengers, local inflammatory cues, through protein-protein interactions, and as imparted by novel gain-of-function mutations. High-resolution TRPA1 structures exist in the open and closed states, which are sampled during normal channel activity. While they represent a major advance, these structures do not address the fundamental question of how TRPA1 becomes sensitized to confer channel hyperactivity in disease. Understanding these mechanisms would open the door to develop new targeted therapeutics. During the next 5 years, we will use complementary biochemical, biophysical, and structural biology approaches to determine the molecular basis of channel hyperactivity conferred by a novel gain-of-function TRPA1 mutation. Our preliminary data suggest this TRPA1 mutant protein co-assembles with wild type TRPA1 subunits to form hyperactive channels. We want to understand how the structural alterations introduced by this TRPA1 mutant affect channel function. Additionally, we will determine how TRPA1 is sensitized and/or activated by calcium and calmodulin. Many local inflammatory cues indirectly activate TRPA1 downstream of G-protein coupled receptors that promote intracellular calcium release. TRPA1 could bind calcium directly or the universal calcium sensor, calmodulin could mediate calcium sensing. Our preliminary data support an interplay of calcium and calmodulin in regulating TRPA1 and we want to understand how calmodulin binding works in concert with calcium-binding sites to confer TRPA1 calcium-dependent channel activity. Collectively, this work will enhance our understanding of regulation and dysregulation of TRPA1 at the molecular level and will uncover novel avenues for drug development to target aberrant channels in chronic pain and inflammatory conditions.

项目概要 芥末受体 TRPA1 是一种非选择性同源四聚体阳离子通道，在原代细胞中表达 感觉神经元被有毒化学刺激物激活有助于疼痛感知和 局部炎症。局部炎症信号反过来又使感觉神经元对疼痛刺激敏感。之内 在疼痛和局部炎症调节周期中，TRPA1 充当正调节因子，其 调节失调可能会导致慢性疼痛的发生。小鼠 TRPA1 基因缺失 消除对化学刺激物、机械和热过敏产生的疼痛感 组织损伤和哮喘引起的气道炎症支持该模型。功能获得TRPA1 突变会导致人类先天性疼痛疾病，突显其在疼痛感知中的直接作用。 这使得了解 TRPA1 功能和失调非常重要。基础科学研究 Paulsen 实验室的主要目标是确定 TRPA1 调节的分子机制和 第二信使的失调，局部炎症信号，通过蛋白质-蛋白质相互作用， 以及由新的功能获得性突变所赋予的。高分辨率 TRPA1 结构存在于 打开和关闭状态，在正常通道活动期间采样。虽然他们代表了一个 重大进展，这些结构并没有解决 TRPA1 如何成为的基本问题 敏感以赋予疾病中的通道过度活跃。了解这些机制将打开 开发新的靶向疗法的大门。未来5年，我们将利用互补 生物化学、生物物理和结构生物学方法来确定分子基础 新型功能获得性 TRPA1 突变导致通道过度活跃。我们的初步数据 表明该 TRPA1 突变蛋白与野生型 TRPA1 亚基共同组装形成过度活跃 渠道。我们想要了解 TRPA1 突变体引入的结构改变如何影响 通道功能。此外，我们将确定 TRPA1 如何被钙敏化和/或激活 和钙调蛋白。许多局部炎症信号间接激活 G 蛋白下游的 TRPA1 促进细胞内钙释放的偶联受体。 TRPA1 可以直接结合钙或 通用钙传感器，钙调蛋白可以介导钙传感。我们的初步数据支持 钙和钙调蛋白在调节 TRPA1 中的相互作用，我们想了解钙调蛋白如何 结合与钙结合位点协同作用，赋予 TRPA1 钙依赖性通道 活动。总的来说，这项工作将增强我们对 TRPA1 调节和失调的理解 在分子水平上，并将揭示针对异常通道的药物开发的新途径 在慢性疼痛和炎症条件下。