Mechanisms of opioid- mediated HIV neuropathogenesis

阿片类药物介导的 HIV 神经发病机制

• 批准号: 10612386
• 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 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10612386
• 关键词: Acceleration; American; Astrocytes; Blood - brain barrier anatomy; Brain; Brain Injuries; CCL2 gene; CD14 gene; CD4 Positive T Lymphocytes; Cell Separation; Cells; Central Nervous System Diseases; Central Nervous System Infections; Chronic; Cognitive deficits; Development; Disease; Enhancers; Exposure to; FCGR3B gene; Gene Expression; Genes; Genetic Techniques; Goals; HIV; HIV Infections; HIV Seropositivity; HIV therapy; Heroin; Human; Impaired cognition; In Vitro; Individual; Infection; Inflammation; Inflammatory; Intervention; Length; Macrophage; Macrophage Activation; Mediating; Mediator; Microglia; Modeling; Molecular; Morphine; Mus; National NeuroAids Tissue Consortium; Neuronal Dysfunction; Neurons; Neuropathogenesis; Opioid; Opioid Receptor; Palliative Care; Pathway interactions; Pattern; Peripheral; Persons; Plasma; Process; Production; Productivity; Proteins; Proviruses; Quality of life; Signal Pathway; Substance Use Disorder; Substance abuse problem; Techniques; Therapeutic; Transgenic Mice; Transgenic Organisms; Viral; Viral Proteins; Viremia; Virus; antiretroviral therapy; blood-brain barrier crossing; brain endothelial cell; chemokine; cognitive function; cyclin T1; cytokine; excitotoxicity; human migration; immune activation; improved; in vivo; innovation; migration; monocyte; mouse model; neuroinflammation; opioid abuse; opioid use; opioid user; palliative; peripheral blood; prescription opioid; promoter; single-cell RNA sequencing; synergism; 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.

这项建议是为了研究HIV介导的神经炎症的分子机制。 艺术和鸦片。我们将使用吗啡，因为它会加剧许多感染艾滋病毒的人的炎症和中枢神经系统疾病 人民。HIV感染中枢神经系统会导致慢性炎症，导致50%的人认知障碍 被感染的人。ART不能减轻这种炎症和随后的中枢神经系统损伤。炎症是一种 艾滋病毒疾病的关键过程和限制这一点的治疗方法以及正在进行的中枢神经系统病毒播种必须发展到 提高感染者的生活质量。这一点更加紧迫，因为艾滋病毒阳性者的寿命更长。艾滋病病毒 外周感染后不久进入中枢神经系统，尽管有抗逆转录病毒治疗，但仍在感染细胞内持续存在。艾滋病病毒进入 大脑至少部分是由感染的单核细胞跨血脑屏障迁移而起作用的。 (Bbb)。表达CD14和CD16的成熟单核细胞是HIV神经致病的关键介质。这些 单核细胞有效地感染了艾滋病毒，并为跨越血脑屏障做好了准备。一旦进入中枢神经系统，它们可能会 分化为受感染的巨噬细胞，可持续数年。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/未感染单核细胞迁移、持续炎症和病毒播种 介导大量感染者神经元树突修剪和变性的中枢神经系统 机制还不是很清楚。抗逆转录病毒疗法不会消除携带艾滋病毒的细胞。因此，单核/巨噬细胞 尽管抗逆转录病毒治疗成功，但艾滋病毒早期蛋白的激活和产生仍在继续，导致脑损伤。我们 将描述吗啡、艾滋病毒和抗逆转录病毒药物对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， 有效的单细胞RNA测序方法和转基因小鼠表征潜在的治疗方法 抑制炎症，引导ART疗效。我们将描述吗啡和ART对 HIV感染和未感染的人单核细胞在人血脑屏障模型中的迁移和使用 利用scRNA-seq技术鉴定移行性HIV感染者和HIV暴露者的独特基因 单核细胞存在或不存在吗啡；表征艾滋病毒、抗逆转录病毒药物和/或吗啡对单核细胞的影响 人巨噬细胞的功能以及利用scRNA-seq对个体炎症基因表达的影响 人巨噬细胞；应用HIV转基因小鼠模型评估阿片类药物和HIV的体内影响 移行小鼠单个单核细胞体内表达的炎性基因 BBB，以及这些小鼠常驻的单个脑巨噬细胞/小胶质细胞；并比较 从HIV幼稚和HIV感染者脑中分离的单个巨噬细胞/小胶质细胞的炎症基因 感染者，包括那些使用scRNA-seq接受姑息阿片类药物治疗的人。