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.

该提案旨在研究存在 HIV 的情况下 HIV 介导的神经炎症的分子机制 ART 和阿片类药物。我们将使用吗啡，因为它会加剧许多艾滋病毒感染者的炎症和中枢神经系统疾病 人们。中枢神经系统的 HIV 感染会导致慢性炎症，导致 50% 以上的人出现认知缺陷 感染人群。 ART 无法减轻这种炎症和随后的中枢神经系统损伤。炎症是一种 必须开发艾滋病毒疾病和限制这种情况的治疗方法以及正在进行的中枢神经系统病毒播种的关键过程，以 改善感染者的生活质量。随着艾滋病毒呈阳性的人寿命更长，这一问题变得更加紧迫。艾滋病病毒 外周感染后不久就进入中枢神经系统，尽管进行了抗逆转录病毒治疗，但仍持续存在于受感染的细胞内。 HIV进入 大脑至少部分是由受感染的单核细胞跨血脑屏障的迁移介导的 （BBB）。表达 CD14 和 CD16 的成熟单核细胞是 HIV 神经发病的关键介质。这些 单核细胞有效地感染艾滋病毒并准备好穿过血脑屏障。一旦进入中枢神经系统，他们可能 分化为可持续数年的受感染巨噬细胞。这会导致感染和/或激活 中枢神经系统细胞，包括巨噬细胞和小胶质细胞，导致慢性炎症，其特征是产生 病毒和/或病毒蛋白、细胞因子和趋化因子。趋化因子，特别是 CCL2，增加 外周血感染/未感染的单核细胞的迁移，持续的炎症和病毒播种 中枢神经系统介导大量感染者的神经元树突修剪和变性 机制尚不清楚。 ART 并不能消除携带 HIV 的细胞。因此，单核细胞/巨噬细胞 尽管抗逆转录病毒疗法取得了成功，但艾滋病毒早期蛋白质的激活和产生仍在继续，导致脑损伤。我们 将描述吗啡、HIV 和 ART 对介导单核细胞进入中枢神经系统机制的影响 以及随后的病毒重新播种和神经炎症。我们将使用最先进的体外技术， 单细胞 RNA 测序和转基因小鼠的强大方法来表征潜在的治疗方法 限制炎症并指导 ART 的疗效。我们将描述吗啡和 ART 对 HIV感染和未感染的人类单核细胞在人类BBB模型中的迁移和使用 scRNA-seq 用于识别 HIV 感染者和 HIV 暴露者所表达的独特基因 在存在或不存在吗啡的情况下的单核细胞；描述 HIV、ART 和/或吗啡对 人类巨噬细胞的功能，并使用 scRNA-seq 来研究个体炎症基因的表达 人类巨噬细胞；应用 HIV 转基因小鼠模型评估阿片类药物和 HIV 的体内影响 对来自小鼠的单个单核细胞在体内表达的炎症基因的影响 BBB，以及来自这些小鼠的常驻个体大脑巨噬细胞/小胶质细胞；并比较表达式 从未感染艾滋病毒和艾滋病毒感染者的大脑中分离出的单个巨噬细胞/小胶质细胞的炎症基因 感染者，包括使用 scRNA-seq 接受姑息性阿片类药物治疗的人。