Dissecting inflammasome anatomy: mechanistic studies and potential intervention

解剖炎症体解剖：机制研究和潜在干预

• 批准号: 8618504
• 负责人: Qian Yin
• 金额: $ 11.99万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8618504
• 关键词: Acute; Adaptor Signaling Protein; Anatomy; Area; Award; Behavior; Biochemical; Biological; Biological Assay; Biology; Boston; Caspase-1; Cell Death; Cells; Chronic Disease; Communities; Complex; Consult; Coupled; Data; Development; Diabetes Mellitus; Disease; Educational workshop; Electron Microscopy; Filament; Future; Goals; Gout; Human; Hydrocarbons; Hydrogen Bonding; Immune; In Vitro; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Inflammatory Response; Inherited; Insecta; Interleukin-1; Interleukin-18; Intervention; Invaded; Lead; Learning; Malignant Neoplasms; Mammalian Cell; Medical; Membrane Proteins; Mentors; Metabolic Diseases; Mus; Mutation; Names; Nature; Nucleotides; Organism; Pathway interactions; Pediatric Hospitals; Peptides; Pharmaceutical Preparations; Phase; Positioning Attribute; Process; Protein Engineering; Proteins; Psoriasis; Reaction; Regulation; Research; Resolution; Resources; Sepsis; Signal Transduction; Small Molecule Chemical Library; Solubility; Stress; Structural Biologist; Structure; Surface; System; Techniques; Testing; Therapeutic; Therapeutic Intervention; Training; Work; X-Ray Crystallography; Yin; base; biophysical techniques; cell type; cytokine; drug development; high throughput screening; inhibitor/antagonist; interfacial; medical schools; microbial; mouse model; pathogen; procaspase-1; protein complex; public health relevance; receptor; reconstitution; reconstruction; repaired; screening; skills; small molecule; small molecule libraries; therapeutic development; therapeutic target; tool

DESCRIPTION (provided by applicant): The principal aim of this proposal is to elucidate the structures and activation and regulation mechanisms of the multi-protein complex inflammasomes, and to develop tool compounds that perturb the assembly of the inflammasomes with potentials for future therapeutic interventions. Dr. Qian Yin is currently a research fellow in Boston Children's Hospital and Harvard Medical School with an immediate goal of completing one-year mentored research before establishing herself in a tenure-track academic position. Dr. Yin's long-term goals include independent researches to understand human innate immune and inflammatory systems and their interaction with pathogens and environmental and intrinsic stresses, with the hope of finding amenable therapeutic targets and effective means for immune and inflammatory diseases. Dr. Yin is a well-trained structural biologist specialized in innate immune and inflammatory signaling. The project will be conducted at Boston Children's Hospital in the rich research community of Longwood Medical Area of Boston, with all facilities and resources from neighboring Harvard Medical School available to the candidate. Dr. Yin will be mentored by Dr. Hao Wu in structural and biophysical approaches to elucidate inflammasome structures and co-mentored by Dr. Timothy Mitchison in development of high throughput assays and tool compounds. During the one-year mentored period, Dr. Yin will learn new techniques such as electron microscopy and high throughput screening under the guidance of the mentors, and develop professional skills by attending classes, seminars, and workshops. After that Dr. Yin plans to make the transition to independence with the assistance of the proposed awards. Inflammasomes are multi-protein complexes that launch the first-line inflammatory responses. During the mentored phase Dr. Yin proposes to reconstruct high-resolution electron microscopy (EM) structure of the AIM2 inflammasome, with X-ray crystallography providing detailed interfacial interactions (Aim 1). This period will also see Dr. Yin clarify the working mechanism of p202, a natural AIM2 inflammasome inhibitor. The proposed work builds heavily on the preliminary EM data of in vitro reconstitution of ASC PYD domain, a common adaptor in most inflammasomes. For the independent phase, Dr. Yin will 1) investigate the assembly mechanisms of NLRP3 inflammasome and elucidate the differential requirements among various inflammasomes (Aim 2), and 2) assay structure-based stabilized helical peptides and screen chemical libraries for small molecules that can perturb inflammasome assembly, in the hope to find druggable interfaces and tool compounds with potential therapeutic values (Aim 3). RELEVANCE: Inflammation is a self-protection process that needs to be tightly regulated. Understanding inflammasome activation and regulation will provide new avenues for treatment of many metabolic diseases, e.g. diabetes and various cancers, as well as hereditary autoinflammatory diseases.

描述（由申请人提供）：本提案的主要目的是阐明多蛋白复合物炎症小体的结构、激活和调节机制，并开发扰乱炎症小体组装的工具化合物，并具有未来治疗干预的潜力。尹倩博士目前是波士顿儿童医院和哈佛医学院的研究员，其近期目标是在获得终身教职学术职位之前完成一年的指导研究。尹博士的长期目标包括开展独立研究，了解人类先天免疫和炎症系统及其与病原体、环境和内在应激的相互作用，希望找到治疗免疫和炎症疾病的合适治疗靶点和有效手段。尹博士是一位训练有素的结构生物学家，专门研究先天免疫和炎症信号传导。该项目将在波士顿朗伍德医学区丰富的研究社区的波士顿儿童医院进行，候选人可以使用邻近的哈佛医学院的所有设施和资源。尹博士将在结构和生物物理方法方面得到吴浩博士的指导，以阐明炎症小体结构，并得到 Timothy Mitchison 博士的共同指导，开发高通量检测和工具化合物。在一年的指导期间，尹博士将在导师的指导下学习电子显微镜和高通量筛选等新技术，并通过参加课程、研讨会和工作坊来发展专业技能。此后，尹博士计划在拟议奖项的帮助下过渡到独立。炎症小体是启动一线炎症反应的多蛋白复合物。在指导阶段，Yin 博士建议重建 AIM2 炎性体的高分辨率电子显微镜 (EM) 结构，并利用 X 射线晶体学提供详细的界面相互作用（目标 1）。本期尹博士还将阐明天然AIM2炎症小体抑制剂p202的工作机制。拟议的工作很大程度上建立在 ASC PYD 结构域体外重建的初步 EM 数据之上，ASC PYD 结构域是大多数炎症体中的常见接头。在独立阶段，尹博士将1）研究NLRP3炎症小体的组装机制并阐明各种炎症小体之间的差异要求（目标2），以及2）分析基于结构的稳定螺旋肽并筛选化学文库中的小分子，这些小分子可以扰乱炎症小体组装，希望找到具有潜在治疗价值的可药物界面和工具化合物（目标 3）。 相关性：炎症是一种自我保护过程，需要严格监管。 了解炎症小体的激活和调节将为许多代谢疾病的治疗提供新途径，例如代谢性疾病。糖尿病和各种癌症，以及遗传性自身炎症性疾病。