Mechanisms of opioid- mediated HIV neuropathogenesis

阿片类药物介导的 HIV 神经发病机制

• 批准号: 9919529
• 负责人: Joan Weinberger Berman
• 金额: $ 83.41万
• 依托单位: ALBERT EINSTEIN COLLEGE 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: 2019-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9919529
• 关键词: American; Astrocytes; Blood - brain barrier anatomy; Brain; Brain Injuries; CCL2 gene; CD14 gene; CD4 Positive T Lymphocytes; Cells; Central Nervous System Diseases; Central Nervous System Infections; Chronic; Cognitive deficits; Development; Disease; Enhancers; Exposure to; FCGR3B gene; Functional disorder; Gene Expression; Genes; Genetic Techniques; Goals; HIV; HIV Infections; HIV Seropositivity; HIV antiretroviral; HIV therapy; Heroin; Human; Impaired cognition; In Vitro; Individual; Infection; Inflammation; Inflammatory; Intervention; Length; Macrophage Activation; Mediating; Mediator of activation protein; Microglia; Modeling; Molecular; Morphine; Mus; National NeuroAids Tissue Consortium; Neurons; Neuropathogenesis; Opioid; Opioid Receptor; Palliative Care; Pathway interactions; Pattern; Peripheral; Plasma; Process; Production; Proteins; Proviruses; Quality of life; Seeds; Signal Pathway; Substance Use Disorder; Substance abuse problem; Techniques; Therapeutic; Transgenic Mice; Transgenic Organisms; Viral; Viral Proteins; Viremia; Virus; antiretroviral therapy; brain endothelial cell; chemokine; cognitive function; cyclin T1; cytokine; excitotoxicity; immune activation; improved; in vivo; innovation; macrophage; migration; monocyte; mouse model; neuroinflammation; opioid abuse; opioid use; opioid user; palliative; peripheral blood; prescription opioid; promoter; single-cell RNA sequencing; therapeutic target; tissue culture; transcriptome

This proposal is to examine molecular mechanisms of HIV-mediated neuroinflammation in the presence of ART and opiods. We will use morphine as it exacerbates inflammation and CNS disease in many HIV infected people. HIV infection of the CNS results in chronic inflammation that leads to cognitive deficits in > 50% of infected people. This inflammation and subsequent CNS damage is not mitigated with ART. Inflammation is a key process in HIV disease and therapies to limit this and ongoing CNS viral seeding must be developed to improve the quality of life of infected people. This is even more pressing as HIV positive people live longer. HIV enters the CNS soon after peripheral infection and despite ART, persists within infected cells. HIV entry into the brain is mediated, at least in part, by infected monocyte transmigration across the blood brain barrier (BBB). Mature monocytes expressing CD14 and CD16 are key mediators of HIV neuropathogenesis. These monocytes are productively infected with HIV and primed to cross the BBB. Once within the CNS, they may differentiate into infected macrophages that can persist for years. This leads to infection and/or activation of CNS cells, including macrophages and microglia, resulting in chronic inflammation characterized by production of virus and/or viral proteins, and cytokines, and chemokines. Chemokines, in particular CCL2, increase transmigration of peripheral blood infected/uninfected monocytes, continuing inflammation and viral seeding of the CNS that mediates neuronal dendritic pruning and degeneration in a large number of infected people by mechanisms not well understood. ART does not eliminate cells harboring HIV. Thus, monocyte/macrophage activation, and production of HIV early proteins continue, resulting in brain injury despite successful ART. We will characterize effects of morphine, HIV, and ART on mechanisms that mediate monocyte entry into the CNS and on subsequent viral reseeding and neuroinflammation. We will use state of the art in vitro techniques, the powerful approach of single cell RNA sequencing, and transgenic mice to characterize potential therapeutics to limit inflammation and guide efficacy of ART. We will characterize the impact of morphine and ART on transmigration of HIV infected and uninfected human monocytes across a model of the human BBB and use scRNA-seq to identify unique genes expressed by individual transmigrating HIV-infected and HIV-exposed monocytes in the presence or absence of morphine; characterize the impact of HIV, ART, and/or morphine on the function of human macrophages and, using scRNA-seq, on expression of inflammatory genes by individual human macrophages; apply an HIV transgenic mouse model to evaluate the in vivo impact of opioids and HIV on inflammatory genes expressed in vivo by individual monocytes from the mice that transmigrated across the BBB, and by resident individual brain macrophages/microglia from these mice; and compare expression of inflammatory genes by individual macrophages/microglia isolated from the brains of HIV-naïve and HIV- infected individuals including those from people who were on palliative opioid treatment using scRNA-seq.

该建议是检查在存在下HIV介导的神经炎症的分子机制 艺术和OOPID。我们将使用吗啡会加剧许多感染的HIV感染和中枢神经系统疾病 人们。中枢神经系统的HIV感染导致慢性感染，导致> 50％的认知缺陷 感染人群。这种感染和随后的中枢神经系统损害不会用艺术减轻。炎症是 艾滋病毒疾病的关键过程和限制这一病毒播种的疗法的关键过程必须发展为 改善感染者的生活质量。随着艾滋病毒积极的人的寿命更长，这更加紧迫。艾滋病病毒 周围感染和目的地艺术后不久进入中枢神经系统，持续在感染的细胞内。艾滋病毒进入 大脑至少部分通过在血脑屏障上感染的单核细胞传播来介导 （BBB）。表达CD14和CD16的成熟单核细胞是HIV神经病发生的关键介体。这些 单核细胞有效地感染了HIV，并启动以越过BBB。一旦进入中枢神经系统，他们可能 区分可能持续多年的感染巨噬细胞。这导致感染和/或激活 CNS细胞，包括巨噬细胞和小胶质细胞，导致慢性炎症为特征于产生 病毒和/或病毒蛋白，细胞因子和趋化因子的含量。趋化因子，尤其是CCL2，增加了 外周血液感染/未感染的单核细胞的转移，持续炎症和病毒播种 介导许多受感染者的神经元树突修剪和变性的中枢神经系统 机制不太了解。 ART不会消除带有艾滋病毒的细胞。那是单核细胞/巨噬细胞 激活和艾滋病毒早期蛋白质的产生继续，导致脑损伤绝望的艺术。我们 将表征吗啡，艾滋病毒和艺术对介导单核细胞进入中枢神经系统的机制的影响 以及随后的病毒恢复和神经炎症。我们将在体外技术中使用最新技术， 单细胞RNA测序和转基因小鼠的强大方法表征了潜在的治疗 限制炎症和指导艺术效率。我们将表征吗啡和艺术对 在人类BBB模型中感染和未感染的人类单核细胞的艾滋病毒转移和使用 SCRNA-SEQ鉴定由个人传播HIV感染和HIV暴露的独特基因 单核细胞在存在或不存在吗啡的情况下；表征艾滋病毒，艺术和/或吗啡对 人类巨噬细胞的功能以及使用SCRNA-SEQ对个体炎症基因表达的功能 人类巨噬细胞；应用HIV转基因小鼠模型来评估阿片类药物和HIV的体内影响 关于由小鼠的单个单核细胞在体内表达的炎性基因 BBB，以及这些小鼠的居民单个脑巨噬细胞/小胶质细胞；并比较表达 由从HIV-NOPHE和HIV-的大脑中分离出的个体巨噬细胞/小胶质细胞的炎症基因 感染的个体，包括使用Scrna-Seq进行姑息治疗的人的感染者。