Molecular mechanisms of TRPA1 regulation

TRPA1调节的分子机制

• 批准号: 10275544
• 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/10275544
• 关键词: Acute; 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; drug development; gain of function; gain of function mutation; mutant; new therapeutic target; novel; 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.

项目摘要 山葵受体TRPA 1是一种非选择性的同源四聚体阳离子通道，表达于初级淋巴细胞中。 感觉神经元，其被有毒化学刺激物激活有助于疼痛感知， 局部炎症。局部炎症线索反过来使感觉神经元对疼痛刺激敏感。内 在疼痛和局部炎症调节周期中，TRPA 1作为正调节剂， 失调可能导致慢性疼痛的发展。小鼠中TRPA 1的遗传丢失 消除了对化学刺激物的疼痛感，机械和热超敏反应， 组织损伤和哮喘诱导的气道炎症支持该模型。功能增益TRPA 1 突变导致人类先天性疼痛障碍，突出了其在疼痛感知中的直接作用。 这使得理解TRPA 1功能和失调非常重要。基础科学研究 保尔森实验室的主要目标是确定TRPA 1调节的分子机制， 通过蛋白质-蛋白质相互作用，第二信使，局部炎症线索， 并且是由新的功能获得性突变所赋予的。高分辨率的TRPA 1结构存在于 打开和关闭状态，在正常通道活动期间进行采样。虽然他们代表了一个 虽然这是一个重大进步，但这些结构并没有解决TRPA 1如何成为 致敏以在疾病中赋予通道过度活跃。了解这些机制将打开 开发新的靶向疗法的大门。在未来5年内，我们将利用 生物化学，生物物理学和结构生物学方法来确定的分子基础， 一种新的功能获得性TRPA 1突变引起的通道过度活跃。我们的初步数据 表明该TRPA 1突变蛋白与野生型TRPA 1亚基共组装形成高活性 渠道我们想了解这种TRPA 1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TRPA 1是如何被钙敏化和/或激活的， 和钙调素。许多局部炎症因子间接激活G蛋白下游的TRPA 1 促进细胞内钙释放的偶联受体。TRPA 1可以直接结合钙离子， 钙调素是一种通用的钙传感器，它可以介导钙敏感。我们的初步数据支持 钙和钙调素在调节TRPA 1中的相互作用，我们想了解钙调素 结合与钙结合位点协同作用，赋予TRPA 1钙依赖性通道 活动总的来说，这项工作将提高我们对TRPA 1调节和失调的理解。 在分子水平上，并将揭示药物开发的新途径，靶向异常通道 在慢性疼痛和炎症条件下。