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.

感染和细胞应激可以触发细胞质模式识别受体组装细胞膜。 炎性体复合物，其促进炎性细胞因子IL-1β、IL-18和炎性细胞因子的释放。 诱导热凋亡细胞死亡。炎性反应也持续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。但过度 炎性小体激活可促成或引起与炎性细胞因子相关的衰弱症状。 疾病特别是，NLRP 3炎性体与许多疾病直接相关。它有一个 通过不仅感测感染，而且感测细胞应力和组织损伤的独特位置。即使 NLRP 3炎性小体对于平衡体内平衡和疾病至关重要， 因此，它是新治疗策略的主要靶点，其潜在的分子机制，特别是在 人类巨噬细胞，仍然知之甚少。存在许多人类炎性体组分， 在小鼠中是不存在的，它们对人类健康和疾病的功能贡献甚至更不清楚 而不是更保守的因素。阐明独特的人类反应是我们实验室的主要重点。 先天性免疫受体寡聚化启动炎症宿主反应，包括炎性小体 activation.该提案中概述的研究旨在从机械上解开一种新的NLRP 3 炎性小体激活概念。我们发现了一种新的NLRP 3炎性小体 人巨噬细胞中的一种成分，它与NLRP 3相互作用，但在小鼠和我们的初步研究中不存在。 研究表明，NLRP 3需要这种共传感器来进行寡聚化以及招募 炎性小体适配器此外，NLRP 3及其共传感器对于高效成核是必需的 ASC聚合和半胱天冬酶-1活化。共传感器表型NLRP 3敲除 人类巨噬细胞。值得注意的是，NLRP 3突变驱动的细胞因子释放是绝对必要的， 导致Cryopyrin-Associated Periodic Syndrome（CAPS）。我们提出了两个具体的目标，调查 共传感器在NLRP 3炎性小体组装和激活中的机制和功能 巨噬细胞，以及使这种共传感器能够促进NLRP 3炎性体的分子事件 activation.我们将利用CRISPR/Cas9敲除并恢复野生型和突变型共传感器的表达， 蛋白和表达人共传感器的人源化小鼠，用于研究其体内功能。 我们希望我们的研究将揭示新的分子机制，不仅改变我们目前的 了解防止不适当的NLRP 3炎性小体激活的控制机制， 维持体内平衡和人类健康，以及炎症性疾病中NLRP 3驱动的病理。 我们的研究成果将推动该领域的发展，并将对理解疾病具有重要意义。 用于治疗疾病和开发有益于患者并对人类健康产生积极影响的新疗法。