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.

项目摘要 芥末受体TRPA1是一种非选择性同型阳离子通道 有害化学刺激物激活其激活的感觉神经元有助于疼痛感知和 局部炎症。反过来，局部炎症提示使感觉神经元对疼痛刺激敏感。之内 TRPA1的疼痛和局部炎症调节周期是正调节剂及其 失调可能导致慢性疼痛的发展。小鼠TRPA1的遗传丧失 消除了对化学刺激物，机械和热超敏反应的疼痛感知 组织损伤和哮喘诱导的气道炎症支持该模型。功能收益TRPA1 突变导致先天性疼痛疾病在人类中突出其在疼痛感中的直接作用。 这使得理解TRPA1的功能和失调非常重要。基础科学研究 在Paulsen实验室中，旨在确定TRPA1调控的分子机制和 通过蛋白质 - 蛋白质相互作用，第二使者的失调，局部炎症提示， 正如新型功能增益突变所赋予的。高分辨率TRPA1结构存在于 开放和封闭状态，在正常通道活性期间进行采样。虽然它们代表 主要进步，这些结构并未解决TRPA1如何变为的基本问题 敏感以赋予疾病中的通道多动症。了解这些机制将开放 开发新的靶向治疗剂的门。在接下来的5年中，我们将使用补充 生化，生物物理和结构生物学方法来确定分子基础 新型功能获得的TRPA1突变赋予的通道多动症。我们的初步数据 建议该TRPA1突变蛋白与野生型TRPA1亚基共组装以形成多动态 频道。我们想了解该TRPA1突变体引入的结构变化如何影响 通道功能。此外，我们将确定如何通过钙敏化和/或激活TRPA1 和钙调蛋白。许多局部炎症提示间接激活G蛋白下游的TRPA1 促进细胞内钙释放的耦合受体。 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，我们想了解如何钙调蛋白 结合与钙结合位点共同授予TRPA1钙依赖性通道 活动。总的来说，这项工作将增强我们对TRPA1的调节和失调的理解 在分子水平上，将发现用于靶向异常通道的药物开发的新途径 在慢性疼痛和炎症状态下。