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: 10434855
• 负责人: Sabita Roy
• 金额: $ 37.87万
• 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 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10434855
• 关键词: Acquired Immunodeficiency Syndrome; Animals; Antibiotics; Attention; Attenuated; Bacterial Translocation; Binding; Biological; Brain; Breeding; Cells; Chronic; Complex; 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; 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; 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; gut microbiome; gut microbiota; marenostrin; microbial; microbial composition; microbiota; neurocognitive disorder; neuroinflammation; neuropathology; opioid abuse; opioid exposure; opioid use; pathogen; 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 长期使用阿片类药物与肠道微生物失调、肠道屏障破坏和全身细菌有关 易位。肠道渗漏、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，人们的注意力集中在阐明类似 Toll 的影响上。 小胶质细胞激活和 NLRP3 炎性体组装中的受体（TLR）作为持续性机制 神经炎症和神经认知疾病进展。虽然之前的研究已经调查了 HIV相关毒性蛋白（TAT和gp120）和阿片类药物对炎症小体激活的直接影响 小胶质细胞对肠道衍生产物的作用及其对炎症小体激活的影响知之甚少 以及在艾滋病毒滥用者中普遍存在的神经认知障碍。虽然大多数研究 将内毒素水平和源自革兰氏阴性菌的细菌产物与炎症相关联 HIV 疾病进展，最近的研究清楚地表明革兰氏阳性菌明显富集和流行 与正常健康人相比，艾滋病毒感染者的细菌群落。我们的中央 假设是微生物失调，革兰氏阳性细菌群落主要扩张 导致 G+ 细菌产物作为小胶质细胞 TLR2 上的同源配体的系统性泄漏 导致 TLR2 激活并诱导炎性体成分组装。我们的中心假设是 有以下初步数据支持。在药物滥用的情况下使用艾滋病毒啮齿动物模型，我们 证明 1) a) 肠道细菌易位 b) 全身炎症 c) NLRP3 显着增加 HIV 感染动物的小胶质细胞上的表达在 HIV+ 吗啡中显着加剧 与各自的对照组相比，接受治疗的动物。 2)吗啡诱导的NLRP3表达仅 在 BV2 小胶质细胞中存在 TLR2 同源配体时观察到。我们将检验以下假设：肠道 微生物群免疫脑轴激活脑小胶质细胞中的炎症小体信号通路，导致 在滥用阿片类药物的 HIV 感染患者中，持续的炎症导致 HAND 的神经病理学。 目标 1：我们将使用以下方法确定肠道微生物群在小胶质细胞炎症小体激活中的作用：a) 无菌 小鼠 b) 差别抗生素（革兰氏特异性去除）治疗的小鼠和 c) 革兰氏阳性菌感染 粪肠球菌。我们还将研究动物的性别是否作为小胶质细胞的生物变量 HIV 感染背景下阿片类药物依赖模型中 NLRP3 的激活。目标 2：确定角色 通过将 floxed TLR2 与 使用 Cx3Cr1 Cre 小鼠进行小胶质细胞特异性 cre 表达。目标 3：确定益生菌和 TLR2 拮抗剂减弱小胶质细胞上的 NLRP3 诱导。