Role of Gut Microbiome- Brain Axis in Modulating CNS Inflammasomes in the Neuropathology Produced by Opioid Exposure and HIV

肠道微生物组-脑轴在阿片类药物暴露和 HIV 产生的神经病理学中调节中枢神经系统炎症小体的作用

• 批准号: 10754697
• 负责人: Sabita Roy
• 金额: $ 4.43万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-30 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10754697
• 关键词: Acceleration; Acquired Immunodeficiency Syndrome; Animals; Antibiotics; Attention; Attenuated; Bacterial Translocation; Binding; Biological; Brain; Breeding; Cells; Chronic; Complex; Correlation Studies; Data; Development; Disease; Disease Progression; Drug abuse; Drug usage; Drug user; Endotoxins; Enterococcus faecalis; Extravasation; Firmicutes; Functional disorder; Genes; Germ-Free; Gram-Negative Bacteria; Gram-Positive Bacteria; HIV; HIV Envelope Protein gp120; HIV Infections; HIV-1; Hand; Immune; Immune system; Individual; Infection; Inflammasome; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Response; Lead; Leaky Gut; Ligands; LoxP-flanked allele; Microglia; Modeling; Molecular; Morphine; Mus; Neurocognitive; Neuropathogenesis; Opiate Addiction; Opioid; Pathogen detection; Patients; Pattern; Predisposition; Prevalence; Probiotics; Production; Proteins; Ribosomes; Rodent Model; Role; Signal Pathway; Signal Transduction; TLR2 gene; Testing; Tissues; Toll-like receptors; antagonist; bacterial community; cytokine; drug abuser; dysbiosis; glial activation; gut microbiome; gut microbiota; marenostrin; microbial; microbial composition; microbiota; neurocognitive disorder; neuroinflammation; neuropathology; opioid abuse; opioid exposure; opioid use; probiotic therapy; receptor; sex; substance use; systemic inflammatory response

Abstract/Project Summary Chronic opioid use is associated with gut microbial dysbiosis, gut barrier disruption and systemic bacterial translocation. Gut leakiness, microbial translocation and systemic inflammation are hallmarks of HIV disease progression. In recent years, much attention has been focused towards elucidating the impact of Toll-like receptors (TLRs) in microglial activation and NLRP3 inflammasome assembly as a mechanism for sustained neuroinflammation and neurocognitive disease progression. While previous studies have investigated the direct effect of HIV associated toxic proteins (TAT and gp120) and opioids on inflammasome activation on microglial little is known about the role of gut derived products and their impact on inflammasome activation and neurocognitive disorders that are prevalent in substance using HIV patients. Although most studies correlate endotoxin levels and bacterial products derived from gram-negative bacteria with inflammation and HIV disease progression, recent studies clearly show a distinct enrichment and prevalence of gram positive bacterial communities in HIV infected patients when compared to normal healthy individuals. Our central hypothesis is that microbial dysbiosis with predominant expansion of Gram positive bacterial communities leading to systemic leakage of G+ bacterial products serve as cognate ligands on TLR2 on microglial cells leading to TLR2 activation and induction of inflammasome components assembly. Our central hypothesis is supported by the following preliminary data. Using a rodent model of HIV in the context of drug abuse, we demonstrate 1) Significant increase in a) gut bacterial translocation b) systemic inflammation c) NLRP3 expression on microglial cells in HIV infected animals that are dramatically exacerbated in HIV+ Morphine treated animals when compared to their respective controls. 2) Morphine induced NLRP3 expression is only observed in the presence of TLR2 cognate ligand in BV2 microglial cells. We will test the hypothesis that gut- microbiota immune brain axis activate inflammasome signaling pathways in brain microglial cells leading to sustained inflammation contributing to the neuropathology of HAND in an opioid abusing HIV infected patients. Aim 1: We will establish the role of the gut microbiota in microglial inflammasome activation using a) germ free mice b) differential antibiotic (gram specific depletion)treated mice and c) infection with gram positive Enterococcus Faecalis. We will also investigate if sex of the animals serves as a biological variable in microglia NLRP3 activation in our model of opioid dependence in the context of HIV infection. Aim 2: Determine the role of TLR2 in NLRP3 induction in microglial cells using tissue specific deletion by breeding floxed TLR2 with microglial specific cre expression using Cx3Cr1 Cre mice. Aim 3: Determine if combination of probiotics and TLR2 antagonist attenuate NLRP3 induction on microglial cells.

摘要/项目摘要 慢性阿片类药物使用与肠道微生物营养不良，肠道屏障破坏和全身细菌有关 易位。肠道泄漏，微生物易位和全身炎症是艾滋病毒疾病的标志 进展。近年来，人们一直专注于阐明类似Toll的影响 小胶质细胞激活和NLRP3炎性体组装中的受体（TLR）作为持续的机制 神经炎症和神经认知疾病进展。虽然先前的研究调查了 HIV相关毒物（TAT和GP120）和阿片类药物对炎症体激活的直接影响对 关于肠道衍生产物的作用及其对炎症体激活的影响，小胶质细胞已知 和使用HIV患者在物质中普遍存在的神经认知障碍。虽然大多数研究 相关的内毒素水平和细菌产物来自革兰氏阴性细菌与炎症和 HIV疾病进展，最近的研究清楚地表明了革兰氏阳性的明显富集和流行率 与正常健康个体相比，艾滋病毒感染患者的细菌群落感染。我们的中心 假设是微生物营养不良，革兰氏阳性细菌群落的主要扩张 导致G+细菌产物的全身泄漏作为小胶质细胞上TLR2上的同源配体 导致TLR2激活和诱导炎症组成分组件的诱导。我们的中心假设是 由以下初步数据支持。在药物滥用的背景下，使用啮齿动物的艾滋病毒模型，我们 证明1）a）肠道细菌易位的显着增加b）全身炎症c）NLRP3 在HIV+吗啡中严重恶化的HIV感染动物中的小胶质细胞上的表达 与各自对照相比，治疗动物。 2）吗啡诱导的NLRP3表达仅是 在BV2小胶质细胞中的TLR2同源配体存在下观察到。我们将测试肠道的假设 微生物群免疫脑轴激活脑小胶质细胞中的炎性体信号通路，导致 持续的炎症导致阿片类药物感染患者的手部神经病理学。 AIM 1：我们将使用a）无胚膜来确定肠道微生物群在小胶质细胞炎症体激活中的作用 小鼠b）治疗的小鼠差异抗生素（革兰氏特异性耗竭）和c）革兰氏阳性感染 肠球菌粪便。我们还将调查动物的性别是否充当小胶质细胞的生物变量 在我们的阿片类药物依赖模型中，NLRP3激活在HIV感染的情况下。目标2：确定角色 通过与组织特异性缺失在小胶质细胞中NLRP3诱导中的TLR2的of of tlr2诱导，通过将Floxed TLR2与 使用CX3CR1 CRE小鼠的小胶质细胞特异性CRE表达。目标3：确定益生菌和 TLR2拮抗剂减弱了小胶质细胞上的NLRP3诱导。