Role of AMP-activated protein kinase in bacterial endophthalmitis

AMP 激活蛋白激酶在细菌性眼内炎中的作用

• 批准号: 9899998
• 负责人: Ashok Kumar
• 金额: $ 38.35万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9899998
• 关键词: Ablation; Adenosine Monophosphate; Adopted; Adoptive Transfer; Aging; Anti-Inflammatory Agents; Area; Bacteria; Bacterial Antibiotic Resistance; Bioenergetics; Blindness; Bone Marrow; Cells; Characteristics; Chemicals; Chimera organism; Citric Acid Cycle; Complication; Cyclic AMP-Dependent Protein Kinases; Development; Disease; Disease Progression; Endophthalmitis; Energy Metabolism; Event; Exhibits; Eye Infections; Functional disorder; Genes; Genus Hippocampus; Glycolysis; Goals; Host Defense; Human; Immune; Immune response; Infection; Infiltration; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Response; Interleukin-1 beta; Interleukin-10; Interleukin-6; Investigation; Knock-out; Knockout Mice; Lead; Light; Link; Mediating; Mediator of activation protein; Metabolic; Metabolism; Microglia; Mitochondria; Modeling; Modification; Molecular; Mus; Myeloid Cells; Natural Immunity; Nature; Nitric Oxide; Operative Surgical Procedures; Oxidative Phosphorylation; PTGS2 gene; PTPRC gene; Pathogenesis; Pathology; Pathway interactions; Pharmacology; Phase; Phenotype; Phosphorylation; Population; Prevalence; Prevention; Production; Protein Kinase; Reporting; Resolution; Retina; Ribonucleosides; Role; STK11 gene; Source; Staphylococcus aureus; Systems Biology; TNF gene; Testing; Therapeutic; Time; Tissues; Toll-like receptors; Trauma; Vision; Visual; Wild Type Mouse; adduct; antimicrobial; bacterial endophthalmitis; cell growth regulation; cell injury; cytokine; disability; immunomodulatory therapies; intravitreal injection; lipid biosynthesis; macrophage; metabolomics; microbial; mitochondrial dysfunction; monocyte; mouse model; neutrophil; new therapeutic target; novel therapeutic intervention; novel therapeutics; pathogen; pathogenic bacteria; protective effect; response; tissue repair; trafficking; transcriptome; upstream kinase

Project Summary The role of myeloid cells such as neutrophils in providing host defense to microbial infections is well- established; however, the contribution of monocytes/macrophages (Mϕ) to the pathophysiology of bacterial endophthalmitis is less clear. Our preliminary studies revealed that Mϕ depletion results in increased inflammatory mediators at the resolution phase, suggesting their involvement in the resolution of endophthalmitis. The Mϕ perform multiple tasks, including sensing pathogens, tissue repair, and, in response to host-derived mediators, they differentiate into distinct functional phenotypes; a feature termed "plasticity". The "classically activated" Mϕ (M1) produce inflammatory cytokines and nitric oxide, contributing to host tissue damage. Conversely, the "alternatively activated" Mϕ (M2) mediate tissue repair through the elimination of damaged cells/tissue and the production of anti-inflammatory molecules to resolve inflammation. Therefore, understanding the mechanisms governing the phenotypic switch of Mϕ can be utilized to develop novel therapeutic strategies. Our transcriptome and metabolomics analyses of the bacteria-infected retina directed us to the identification of adenosine monophosphate-activated protein kinase (AMPK), a metabolic gene, which modulates the infiltrating myeloid cell phenotype in endophthalmitis. We discovered that mice with global deletion (knockout) of AMPKα1 (KO) developed severe endophthalmitis and pathology compared to wild type (WT) mice. Mϕ lacking AMPKα1 maintained a low metabolic state, even in the hyper-inflammatory state. To precisely examine the role of AMPK in myeloid cells, we induced endophthalmitis in myeloid cell specific KO of AMPKα1 (LysM-KO) and observed that LysM-KO displayed exacerbated inflammation and reduced retinal function compared to WT mice, suggesting an essential role of AMPK in myeloid cells in the pathogenesis of bacterial endophthalmitis. Building on these findings, we propose to test our central hypothesis that AMPK exerts protective effects in bacterial endophthalmitis by modulating the polarization of infiltrating monocytes/Mϕ to promote inflammation resolution and that metabolic reprograming is an underlying mechanism of the monocytes/Mϕ phenotype switch. To test our hypothesis, in Aim 1, we will investigate the mechanisms underlying reduced AMPK activity in bacterial endophthalmitis by examining the modification of LKB1 via nitrosylation or chemical adduct formation. Aim 2 tests the hypothesis that AMPKα1 ablation enhances the activation state of myeloid cells and maintains their proinflammatory (M1) state during the resolution phase of the disease. In Aim 3, we will decipher the bioenergetic events, regulated by AMPK in Mϕ, that polarize and maintain their pro-inflammatory nature. The anticipated results of this study will demonstrate that defective AMPK activity in myeloid cells, mainly in monocytes/M impacts the resolution of endophthalmitis via regulation of cellular metabolism. Also, it may provide novel therapeutic targets for the development of anti- inflammatory therapies for endophthalmitis and other microbial infections.

项目摘要 髓样细胞（例如中性粒细胞）在为微生物感染提供宿主防御方面的作用是很好的 已确立的;然而，单核细胞/巨噬细胞（Mϕ）对细菌的病理生理的贡献 内膜炎不太清楚。我们的初步研究表明，M ϕ部署导致增加 在分辨率阶段的炎症介体，表明它们参与了分辨率 内脑膜。 M ϕ执行多个任务，包括感测病原体，组织修复以及响应 对于宿主衍生的介体，它们分为不同的功能表型。一个称为“可塑性”的功能。 “经典活化”的M ϕ（M1）产生炎症细胞因子和一氧化氮，有助于宿主组织 损害。相反，“替代激活”的M ϕ（M2）通过消除来介导组织修复 受损的细胞/组织和抗炎分子的产生以解决注射。所以， 了解管理M ϕ的表型转换的机制可以用于开发新颖的机制 治疗策略。我们针对细菌的视网膜的转录组和代谢组学分析 美国鉴定腺苷单磷酸激活的蛋白激酶（AMPK），一种代谢基因，该基因 调节内咽中浸润的髓样细胞表型。我们发现与全球的老鼠 与野生型相比 （wt）小鼠。缺乏AMPKα1的M ϕ保持低代谢状态，即使处于过度炎症状态。到 精确地检查了AMPK在髓样细胞中的作用，我们诱导了髓样细胞在特异性KO中的内脑炎 AMPKα1（LYSM-KO），并观察到Lysm-KO显示出加重的注射和视网膜的减少 与WT小鼠相比，功能表明AMPK在髓样细胞中的重要作用在 细菌性内膜。在这些发现的基础上，我们建议测试我们的中心假设，即AMPK 通过调节浸润单核细胞的极化/M ϕ的极化，对细菌发挥保护作用 促进注射分辨率和代谢重编程是 单核细胞/M ϕ表型开关。为了检验我们的假设，在AIM 1中，我们将研究机制 通过检查LKB1通过 硝基化或化学加合物形成。 AIM 2检验了AMPKα1消融的假设增强了 髓样细胞的激活状态并在分辨率阶段保持促炎（M1）状态 疾病。在AIM 3中，我们将破译由MDACH中AMPK调节的生物能量事件，该事件极化和 保持他们的促炎本质。这项研究的预期结果将证明有缺陷 髓样细胞中的AMPK活性，主要是单核细胞/M影响内脑膜的分辨率通过 调节细胞代谢。此外，它可能为抗抗 - 内度炎和其他微生物感染的炎症疗法。