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

基本信息

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
项目类别：
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&apos 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

项目摘要

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.

项目概要这一更新应用是我们长期努力的成果，旨在了解 IKK 依赖性 NF-κB 信号传导如何发挥作用控制炎症和免疫的重点是 NLRP3 炎症小体的正向和负向调节我们的努力将集中于彻底阐明一种新的机制。我们实验室确定的信号传导机制，通过该机制，Toll 样受体 (TLR) 的参与使巨噬细胞 (M) 对压力、损伤信号和触发 NLRP3 炎性体的微粒有反应 NLRP3 炎症小体持续激活并诱导 IL-1β 和 IL-18 产生。多种神经退行性疾病、代谢性疾病和炎症性疾病，例如阿尔茨海默病、II 型糖尿病和骨关节炎 (OA)，但对其了解甚少，阻碍了新型 NLRP3 特异性抗骨关节炎的开发此外，先前尝试通过靶向 IKK 依赖性 NF-来减轻炎症。通过研究 NF-κB 如何抑制 NLRP3 炎症小体的激活，κB 信号传导失败。调节 NLRP3 炎症小体，我们确定了线粒体在控制炎症小体中的关键作用 NF-κB 和 p62 依赖性线粒体自噬终止 NLRP3 炎症小体的激活。受刺激的 M、TLR4 或 TLR3 接合通过基于激活的新颖途径触发 mtDNA 复制 IRF-1 的产生和核苷酸激酶 CMPK2 的诱导该途径对于氧化 (Ox) 的产生至关重要。暴露于多种 NLRP3 炎性体激活剂（例如 ATP、尼日利亚霉素、我们的结果表明，Ox-mtDNA 是 NLRP3 的最终配体。我们将继续研究该途径并研究其适用性。其目标是治疗目前无法治愈的炎症性疾病，例如 OA。确定 CMPK2 敲除和敲入小鼠是否表现出有缺陷的 NLRP3 炎性体激活和因此，我们还将确定 CMPK2- 如何抵抗羟基磷灰石引起的关节炎症。依赖的 mtDNA 复制支持 Ox-mtDNA 的产生并检查后者是否结合 NLRP3 直接绘制结合位点图谱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 的结合，我们将研究如何结合。其他 mt 信号和代谢物、活性氧 (ROS) 和衣康酸 (IA)，调节合成 pro-IL-1β、pro-IL-18 和其他细胞因子将集中于氧化剂反应性的作用。转录因子 NRF2 在细胞因子基因表达中的作用，还将探讨 mtROS 和 IA 如何影响 NF-κB 这些研究将扩展我们对 NRF2 的基本理解。控制炎症的机制将为开发新型抗炎药物奠定基础抑制 IL-1β 和 IL-18 的产生而不干扰抗菌免疫。