NF-kappaB and Mitochondrial Signals as Positive and Negative Regulators of Inflammation

NF-kappaB 和线粒体信号作为炎症的正向和负向调节剂

• 批准号: 10266224
• 负责人: Michael Karin
• 金额: $ 3.38万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-29 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10266224
• 关键词: Ablation; Acids; Affect; Agonist; Alzheimer' s Disease; Anti-Inflammatory Agents; Attenuated; Autophagocytosis; Binding; Binding Sites; Biochemical; Biological; Cardiovascular Diseases; Citric Acid Cycle; Cysteine; Cytidine Monophosphate; Cytokine Gene; DNA biosynthesis; DNA metabolism; Degenerative Disorder; Degenerative polyarthritis; Dementia; Dependence; Development; Disease; Drug Targeting; Ensure; Enzymes; Event; Exhibits; Exposure to; Foundations; Gene Expression; Generations; Genetic Transcription; Goals; Gouty Arthritis; Hydroxyapatites; IRF1 gene; Immunity; Inflammasome; Inflammation; Inflammatory; Inflammatory Response; Interleukin-1 beta; Interleukin-10; Interleukin-18; Knock-in Mouse; Knockout Mice; Laboratories; Ligands; Liposomes; Maps; Mediating; Metabolic; Mitochondria; Mitochondrial DNA; NF-kappa B; Nerve Degeneration; Nigericin; Non-Insulin-Dependent Diabetes Mellitus; Nucleotides; Oxidants; Oxides; Pathogenesis; Pathogenicity; Pathology; Pathway interactions; Pharmaceutical Preparations; Phosphotransferases; Process; Production; Prostaglandins; Reactive Oxygen Species; Receptor Signaling; Regulation; Research; Resistance; Role; Scaffolding Protein; Signal Transduction; Sterility; Stress; Structure; TLR3 gene; TLR4 gene; Testing; Tissues; Toll-like receptors; Uridine Kinase; adduct; age related; aluminum sulfate; antimicrobial; base; cyclopentanone; cytokine; experimental study; in vitro Assay; in vivo; joint inflammation; knock-down; macrophage; mitochondrial metabolism; novel; nuclease; oxidation; prevent; regenerative; repair enzyme; response; therapeutic target; tissue injury; tissue repair; transcription factor

PROJECT SUMMARY This renewal application of our long-term effort to understand how IKK-dependent NF-κB signaling controls inflammation and immunity is focused on positive and negative regulation of the NLRP3 inflammasome by NF-κB and mitochondrial (mt) metabolism. Our effort will be placed on complete elucidation of a novel signaling mechanism, identified in our laboratory, through which engagement of Toll-like receptors (TLR) renders macrophages (M) responsive to stress, damage signals and microparticles that trigger NLRP3 inflammasome activation and induce IL-1β and IL-18 production. Persistent NLRP3 inflammasome activation is involved in several neurodegenerative, metabolic and inflammatory diseases, e.g. Alzheimer’s disease, type II diabetes and osteoarthritis (OA), but its poor understanding has prevented development of novel NLRP3-specific anti- inflammatory drugs. Furthermore, previous attempts to alleviate inflammation by targeting IKK-dependent NF- κB signaling have failed due to enhanced NLRP3 inflammasome activation. By studying how NF-κB negatively regulates the NLRP3 inflammasome, we identified a critical role for mitochondria in control of inflammasome activity. Whereas, NF-κB- and p62-dependent mitophagy terminates NLRP3 inflammasome activation in stimulated M, TLR4 or TLR3 engagement triggers mtDNA replication via a novel, pathway based on activation of IRF-1 and induction of the nucleotide kinase CMPK2. This pathway is essential for production of oxidized (Ox) mtDNA in TLR-activated M that were exposed to diverse NLRP3 inflammasome activators, e.g. ATP, nigericin, alum and DOTAP liposomes. Our results suggest that Ox-mtDNA is the ultimate NLRP3 ligand responsible for inflammasome assembly and activation. We will continue to study this pathway and investigate the suitability of its targeting for treatment of currently incurable inflammatory diseases, such as OA. Accordingly, we will determine whether CMPK2 knockout and knockin mice exhibit defective NLRP3 inflammasome activation and are therefore resistant to hydroxyapatite-induced joint inflammation. We will also determine how CMPK2- dependent mtDNA replication supports Ox-mtDNA production and examine whether the latter binds NLRP3 directly, map the binding site and conduct biochemical and structural studies to determine how Ox-mtDNA binding induces the association of NLRP3 with the inflammasome scaffold protein ASC. We will investigate how other mt signals and metabolites, reactive oxygen species (ROS) and itaconic acid (IA), modulate synthesis of pro-IL-1β, pro-IL-18 and other cytokines. These studies will focus on the role of the oxidant-responsive transcription factor NRF2 in cytokine gene expression and will also explore how mtROS and IA affect NF-κB activity and its crosstalk with NRF2. These studies will expand our basic understanding of the fundamental mechanisms that control inflammation and will lay the foundation for developing novel anti-inflammatory drugs that inhibit IL-1β and IL-18 production without interfering with antimicrobial immunity.

项目摘要 我们长期努力的这种更新应用于了解IKK依赖性NF-κB信号传导 控制炎症和免疫力集中于NLRP3炎症体的正和负调节 由NF-κB和线粒体（MT）代谢。我们的努力将全面阐明小说 信号传导机制，在我们的实验室中确定，通过该机制，Toll样受体（TLR）的参与度呈现 巨噬细胞（M）响应压力，损害信号和触发NLRP3炎性体的微粒 激活并诱导IL-1β和IL-18产生。持续的NLRP3炎性体激活参与 几种神经退行性，代谢和炎症性疾病，例如阿尔茨海默氏病，II型糖尿病和 骨关节炎（OA），但其不良的理解阻止了新型NLRP3特异性抗 - 炎症药。此外，以前试图通过靶向IKK依赖性NF-来减轻注射 由于NLRP3炎性体激活的增强，κB信号传导失败了。通过研究NF-κB如何负面 调节NLRP3炎性体，我们确定了线粒体控制炎症体的关键作用 活动。而NF-κB-和p62依赖性线粒体终止NLRP3炎性体激活 通过基于激活的小说，刺激M，TLR4或TLR3参与触发mtDNA复制 IRF-1和核苷酸激酶CMPK2的诱导。该途径对于生产氧化（OX）至关重要 TLR激活的m中的mtDNA暴露于潜水员NLRP3炎性体激活剂，例如aTP，尼古丁， 明矾和DOTAP脂质体。我们的结果表明，ox-mtDNA是负责的最终NLRP3配体 炎症体组装和激活。我们将继续研究这一途径，并调查 它的目标是治疗当前发生的炎症性疾病，例如OA。根据，我们会 确定CMPK2敲除和敲除小鼠是否暴露了有缺陷的NLRP3炎性体激活和 因此，对羟基磷灰石诱导的关节注射具有抗性。我们还将确定CMPK2-如何 依赖的mtDNA复制支持OX-MTDNA的产生和检查后者是否结合NLRP3 直接绘制结合位点并进行生化和结构研究，以确定OX-MTDNA如何 结合诱导NLRP3与炎性体支架蛋白ASC的缔合。我们将调查如何 其他MT信号和代谢产物，活性氧（ROS）和Itaconic酸（IA），调节合成的合成 Pro-IL-1β，Pro-IL-18和其他细胞因子。这些研究将集中于氧化物响应的作用 转录因子NRF2在细胞因子基因表达中，还将探索MTROS和IA如何影响NF-κB 活动及其与NRF2的串扰。这些研究将扩大我们对基本的基本理解 控制注射并将为开发新型抗炎药奠定基础的机制 抑制IL-1β和IL-18的产生而不会干扰抗菌免疫。