A novel mechanism for NLRP3 inflammasome activation in human macrophages

人类巨噬细胞中 NLRP3 炎症小体激活的新机制

• 批准号: 10343393
• 负责人: Andrea Dorfleutner
• 金额: $ 76.57万
• 依托单位: CEDARS-SINAI MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-16 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10343393
• 关键词: ATP phosphohydrolase; Address; Affect; Alzheimer' s Disease; Asbestosis; Atherosclerosis; Binding; Biochemical; Biological; CASP1 gene; CRISPR/Cas technology; Cell Death; Cells; Cellular Stress; Complex; Development; Diabetes Mellitus; Disease; Equilibrium; Event; Gout; Health; Homeostasis; Human; Immune; Immune response; Immunologic Receptors; Immunology; Individual; Infection; Infectious Agent; Inflammasome; Inflammatory; Inflammatory Response; Interleukin-1 beta; Interleukin-18; Interleukins; Knock-out; Knockout Mice; Licensing; Link; Malignant Neoplasms; Mediating; Modeling; Modification; Molecular; Mus; Mutation; Outcome; Pathology; Patients; Pattern; Pattern recognition receptor; Periodicity; Phenocopy; Positioning Attribute; Proteins; Regulation; Research; Role; Sensory; Signal Transduction; Silicosis; Sterility; Syndrome; Tissues; associated symptom; base; cytokine; design; disabling symptom; humanized mouse; improved; in vivo; inducible gene expression; kidney dysfunction; knockout gene; macrophage; marenostrin; mouse genetics; mutant; novel; novel therapeutics; pathogen; pathogenic microbe; polymerization; prevent; receptor; recruit; response; sensor; stress granule; tool; treatment strategy; wound healing

Infections and cellular stress can trigger cytoplasmic pattern recognition receptors to assemble an inflammasome complex, which promotes the release of the inflammatory cytokines IL-1β, IL-18 and the induction of pyroptotic cell death. Inflammasome responses are also perpetuated and propagated to bystander cells. Ultimately, this response contributes to pathogen clearance and wound healing. However, excessive inflammasome activation can contribute to- or cause debilitating symptoms associated with inflammatory diseases. Particularly, the NLRP3 inflammasome has been directly linked to numerous diseases. It has a unique position by not only sensing infections, but also cellular stress and tissue damage. Even though the NLRP3 inflammasome is of utmost importance for balancing between homeostasis and disease, and is therefore a prime target for novel treatment strategies, the underlying molecular mechanisms, particularly in human macrophages, are still poorly understood. There are numerous human inflammasome components that are absent in mice and their functional contribution to human health and disease are even less well understood than the more conserved factors. Elucidating unique human responses is the main focus of our lab. Innate immune receptor oligomerization initiates inflammatory host responses, including inflammasome activation. The research outlined in this proposal is designed to mechanistically unravel a novel NLRP3 inflammasome activation concept in human macrophages. We discovered a novel NLRP3 inflammasome component in human macrophages, which interacts with NLRP3, but is absent from mice and our preliminary studies revealed that NLRP3 requires this co-sensor for oligomerization as well as for recruiting the inflammasome adaptor, ASC. Furthermore, NLRP3 and its co-sensor are necessary for efficiently nucleating ASC polymerization and caspase-1 activation. Knock out of the co-sensor phenocopies NLRP3 knock out in human macrophages. Significantly, it is absolutely necessary for cytokine release driven by NLRP3 mutations that cause Cryopyrin-Associated Periodic Syndrome (CAPS). We propose two specific aims that investigate the mechanism and function of the co-sensor in NLRP3 inflammasome assembly and activation in macrophages, as well as the molecular events that enable this co-sensor to promote NLRP3 inflammasome activation. We will utilize CRISPR/Cas9 knock out and restored expression of wild type and mutant co-sensor proteins and a humanized mouse expressing the human co-sensor for studying its function in vivo. We expect that our research will uncover novel molecular mechanisms that not only change our current understanding of control mechanisms that prevent inappropriate NLRP3 inflammasome activation for maintaining homeostasis and human health, but also NLRP3-driven pathologies in inflammatory diseases. The outcomes of our study will move the field forward and will be highly significant for understanding disease pathologies and for the development of novel therapies that benefit patients and positively affect human health.

感染和细胞应激会触发细胞质模式识别受体，以组装 炎性体复合物，促进炎性细胞因子IL-1β，IL-18和 诱导凋亡细胞死亡。炎性组的反应也会永久存在并传播给旁观者 细胞。最终，这种反应有助于病原体清除和伤口愈合。但是，超过 炎性体激活会导致或引起与炎症有关的衰弱症状 疾病。特别是，NLRP3炎性体已与许多疾病直接相关。它有一个 独特的位置不仅是感染感染，还通过细胞应力和组织损伤。即使 NLRP3炎性体对于平衡体内平衡和疾病至关重要，并且是 因此，是新型治疗策略的主要目标，是基本的分子机制，特别是 人类巨噬细胞仍然很少了解。有许多人类炎症组成部分 小鼠不存在，它们对人类健康和疾病的功能贡献甚至不太了解 比更保守的因素。阐明独特的人类反应是我们实验室的主要重点。 先天免疫受体寡聚启动炎症宿主反应，包括炎症体 激活。该提案中概述的研究旨在从机械上解开新的NLRP3 人类巨噬细胞中的炎症体激活概念。我们发现了一个新颖的NLRP3炎症体 与NLRP3相互作用的人类巨噬细胞中的成分，但不存在小鼠和我们的初步 研究表明，NLRP3需要此共同传感器进行寡聚以及招募 炎症适配器，ASC。此外，NLRP3及其共同传感器对于有效成核是必需的 ASC聚合和caspase-1激活。从共同传感器的表演中敲出NLRP3敲门 人类巨噬细胞。值得注意的是，由NLRP3突变驱动的细胞因子释放绝对必要 这会导致冷冻蛋白相关的周期性综合征（CAPS）。我们提出了两个调查的具体目标 NLRP3炎性组组装中共同传感器的机理和功能和激活中的机理和功能 巨噬细胞以及能够促进NLRP3炎性体的分子事件 激活。我们将利用CRISPR/CAS9敲除并恢复野生型和突变共同传感器的表达 蛋白质和人源化的小鼠表达人类共同传感器，用于研究其在体内的功能。 我们预计我们的研究将发现新颖的分子机制，不仅改变了我们的当前 理解控制机制，以防止不适当的NLRP3炎性体激活 维持体内稳态和人类健康，以及炎症性疾病中NLRP3驱动的病理。 我们研究的结果将向前进，对于理解疾病非常重要 病理学和开发新的疗法，使患者受益并积极影响人类健康。