Molecular mechanisms of gasdermin recognition by proteases and autophagy proteins in cytokine release

细胞因子释放中蛋白酶和自噬蛋白识别gasdermin的分子机制

• 批准号: 10223159
• 负责人: Tsan Sam Xiao
• 金额: $ 40.25万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-24 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10223159
• 关键词: Alzheimer' s Disease; Apoptosis; Autoimmune; Autophagocytosis; Binding Sites; CASP1 gene; CASP3 gene; CASP8 gene; Caspase; Caspase Inhibitor; Catalytic Domain; Cell Death; Cell membrane; Cells; Collaborations; Complex; Crystallization; Cytolysis; Data; Data Analyses; Disease; Epithelial Cells; Event; Experimental Designs; Goals; Human; Immune; Immune signaling; Immunity; Infection; Inflammasome; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Interleukin-1 beta; Interleukin-18; Investigation; Knowledge; Lead; Length; Lipid Binding; Lytic; Masks; Mediating; Membrane; Molecular; Molecular Chaperones; Multiple Sclerosis; Mus; Neutrophil Activation; Nonlytic; Outcome; Peptide Hydrolases; Peptide Signal Sequences; Play; Proteins; Proteomics; Regulation; Reporting; Role; Serine Protease; Signal Pathway; Signal Transduction; Site; Structure; T-Lymphocyte; Testing; Tissues; Vesicle; autoinflammatory; base; cell type; cytokine; enzyme substrate; enzyme substrate complex; insight; macrophage; mast cell; membrane assembly; neutrophil; novel; pathogen; programs; receptor; recruit; repaired; success; therapeutic development; therapeutic target

Abstract The goal of this proposal is to investigate mechanisms for pyroptosis-dependent and novel pyroptosis- independent export of proinflammatory cytokines mediated by caspases, secretory autophagy proteins, and gasdermin D (GSDMD), a recently identified effector molecule for pyroptosis. The inflammasomes are crucial innate immune signaling platforms implicated in immune defense against infections and autoimmune/ autoinflammatory disorders such as inflammatory bowel disease, multiple sclerosis, and Alzheimer’s disease. Activation of the inflammasome signaling pathways leads to the maturation and secretion of cytokines such as IL-1β and IL-18, and an inflammatory form of programmed cell death in certain cell types called pyroptosis. GSDMD assembles membrane pores upon cleavage by inflammatory caspases-1, 4, 5 and 11 and caspase-8. In macrophages, such pores allow the release of mature cytokines upon cytolysis. Intriguingly, pyroptosis- and membrane pore-independent function of GSDMD during secretion of IL-1β upon inflammasome activation has been demonstrated for neutrophils, epithelial cells and T cells (Projects 1, 2, and 4), in which GSDMD is recruited by secretory autophagy proteins to facilitate a novel non-lytic form of cytokine release. Specific recognition of GSDMD by caspases or autophagy proteins thus underlie GSDMD function in lytic or non-lytic cytokine release. We hypothesize that inflammatory caspases recognize GSDMD through distinct protein-protein interfaces during lytic cytokine release. By contrast, GSDMD is recruited by autophagy machinery to facilitate novel non-lytic cytokine secretion independent of membrane pore formation. We will focus on the following specific aims in a collaborative Program Project to test the above hypothesis. Aim 1. Define the mechanisms of protease- dependent activation of GSDMD in lytic release of cytokines. We propose to elucidate the mechanisms of GSDMD recognition by caspases and serine proteases in collaboration with Projects 1, 2 and 4. Discovery from our proposal may facilitate investigation into specific caspase inhibitors based on distinct enzyme-substrate interfaces, which may target pyroptosis, apoptosis and other inflammatory signaling pathways involving caspases. Aim 2. Define the mechanisms of GSDMD-guided non-lytic secretion of cytokines. We propose to characterize the recruitment of IL-1β and GSDMD by chaperones and autophagy proteins. The roles of these interaction in non-lytic cytokine release will be probed in collaboration with Projects 2 and 4. The success of this project will reveal the molecular mechanisms for the lytic or non-lytic secretion of proinflammatory cytokines mediated by GSDMD. This proposal draws on the strength of the other three Projects to facilitate and validate our structural studies. Structural insights from our studies will in turn inform experimental design and data interpretation for the other Projects. As such, outcomes from the four Projects benefit each other as an interdependent and integrated Program.

抽象的 该提案的目的是研究依赖性和新颖的凋亡的机制 由胱天蛋白酶，秘书自噬蛋白和 Gasdermin D（GSDMD），最近鉴定出的用于凋亡的效应分子。炎症是至关重要的 针对感染和自身免疫的免疫防御中实施的先天免疫信号平台/ 自身炎症性疾病，例如炎症性肠病，多发性硬化症和阿尔茨海默氏病。 炎性体信号通路的激活导致细胞因子的成熟和分泌，例如 IL-1β和IL-18，以及某些称为凋亡的细胞类型的程序性细胞死亡的炎症形式。 GSDMD通过炎症caspase-1、4、5和11和caspase-8裂解后会组装膜孔。 在巨噬细胞中，这种毛孔允许在细胞溶解后释放成熟的细胞因子。有趣的是，凋亡 - 和 GSDMD在炎症体激活后分泌IL-1β期间GSDMD的膜孔依赖性功能 被证明用于中性粒细胞，上皮细胞和T细胞（项目1、2和4），其中募集了GSDMD 由秘书自噬蛋白促进细胞因子释放的新型非散晶形式。具体认可 GSDMD由胱天蛋白酶或自噬蛋白通过裂解或非柳细胞因子释放中的GSDMD功能的基础。 我们假设炎性caspases在期间通过不同的蛋白质 - 蛋白质界面识别GSDMD 裂解细胞因子释放。相比之下，GSDMD由自噬机制招募，以促进新颖的非散热 细胞因子分泌与膜孔形成无关。我们将专注于以下特定目标 协作计划项目以检验上述假设。目标1。定义蛋白酶的机制 GSDMD的依赖性细胞因子释放中的激活。我们建议阐明 与项目1、2和4合作的caspases和串行蛋白的GSDMD识别 我们的建议可能促进基于不同酶 - 基层的特定caspase抑制剂的投资 界面，可能针对凋亡，凋亡和其他炎症信号通路的界面 caspase。目标2。定义GSDMD引导的细胞因子非乳酸分泌的机制。我们建议 表征由伴侣和自噬蛋白募集IL-1β和GSDMD。这些角色 非透明细胞因子释放中的相互作用将与项目2和4合作进行探测。 项目将揭示促炎细胞因子的裂解或非散性分泌的分子机制 由GSDMD介导。该提案借鉴了其他三个项目的实力，以促进和验证 我们的结构研究。我们研究的结构洞察力反过来将为实验设计和数据提供信息 对其他项目的解释。因此，四个项目的结果彼此受益 相互依存和集成程序。