Role of exosome extracellular vesicles in opiate abuse and HIV neuropathogenesis

项目摘要 分泌型细胞外小泡(EVS)可能在生物学过程和疾病发病机制中发挥作用。影响 在这些关于人类免疫缺陷病毒1型(HIV-1)感染的EV中，最近才开始 调查过了。事实上，EV，如外周小体，已经被证明影响中枢神经内的细胞 中枢神经系统(CNS)和调节对病原体的免疫反应。HIV阴性因子(Nef)从以下途径释放 NEF转基因或HIV感染的免疫细胞在外体中，细胞外纳米大小的囊泡通常用于 辅助或细胞间通讯--抗原的递送、基因表达的修饰和基因的调节 免疫反应。有趣的是，感染了HIV的小胶质细胞或转染了nef-GFP表达载体的小胶质细胞 在外体中释放Nef。然而，这种胞外外体Nef(ExNef)可能在HIV复制中发挥作用 在中枢神经系统内，其发病机制尚不清楚。众所周知，艾滋病毒感染大脑内的细胞，并持续存在 在中枢神经系统内，尽管成功地联合抗逆转录病毒治疗(CART)，并引起神经认知 损害，如艾滋病毒相关的神经疾病(手)。尽管Cart显著降低了 外周病毒载量达到无法检测到的水平(无血症)，手中仍可观察到40%的病毒被抑制 HIV+的个人。总之，这些发现表明，在CART存在的情况下，一种新的机制不会 与艾滋病毒相关的是导致神经认知障碍的因素。药物滥用也可能起到关键作用 在艾滋病毒疾病进展和神经认知功能障碍的发病中的作用。鸦片类药物，如海洛因，及其 活性代谢物吗啡已被证明可增加艾滋病毒疾病进展为神经艾滋病和 增加艾滋病毒/艾滋病患者手部感染的风险和严重程度。考虑到那几乎是一个- 三分之一的PLWA报告海洛因滥用，了解鸦片类药物和CART如何在HIV中相互作用是很重要的 疾病导致神经认知障碍，以改善这些艾滋病毒感染者的生活质量。我们 假设阿片剂诱导的Nef+Ev组成和释放的改变会加剧exNef 在CART的背景下，相关的神经元损伤并导致更大的神经认知障碍。在……里面 我们将在购物车和鸦片类药物的背景下调查这项提议，细胞外小泡的影响， 特异性地从HIV感染(或nef转基因)神经元上的小胶质细胞释放exNef以了解 人类免疫缺陷病毒致神经认知障碍的机制(S)我们还将 进行一项横断面研究，比较CART上PLWA中脑脊液(CSF)exNef与 历史/当前阿片类药物使用和神经认知障碍/手。这项提案的结果将使我们能够 证明EVS在中枢神经系统中阿片类药物和HIV相互作用引起的神经发病中的作用。

Project Summary Secreted extracellular vesicles (EVs) may play a role in biological processes and disease pathogenesis. Impact of these EVs on Human Immunodeficiency Virus type 1(HIV-1) infection has only recently started to be investigated. In fact, EVs such as exosomes have been shown to influence cells within the central nervous system (CNS) and modulate immune responses to pathogens. The HIV Negative factor (Nef) is released from nef-transfected or HIV-infected immune cells in exosomes, extracellular nano-sized vesicles generally used for para- or intercellular communication – delivery of antigen, modification of gene expression, and modulation of immune responses. Interestingly, microglia infected with HIV or transfected with a nef-gfp expression plasmid release Nef in exosomes. However, the role this extracellular exosomal Nef (exNef) may have in HIV replication within the CNS and neuropathogenesis is unknown. It is known that HIV infects cells within the brain, persists within the CNS despite successful combination anti-retroviral therapy (cART), and causes neurocognitive impairments such as HIV-associated Neurological Disorder (HAND). Although cART significantly lowers peripheral viral load to undetectable levels(aviremia), HAND is still observed in among 40% of virally suppressed HIV+ individuals. Together these findings suggest that in the presence of cART a novel mechanism not associated with the HIV is at play to induce neurocognitive impairment. Substance abuse could also play a key role in HIV disease progression and the onset of neurocognitive impairment. Opiates such as heroin, and its active metabolite morphine have been shown to increase the rate of HIV disease progression to NeuroAIDS and increase both the risk and severity of HAND in people living with HIV/AIDS (PLWHAs). Given that almost one- third of PLWHAs report heroin abuse, it is important to understand how opiates and cART interplay in HIV disease to cause neurocognitive impairment in order to improve the quality of life for these HIV+ individuals. We hypothesize that opiate-induced modifications in Nef+ EV composition and release exacerbates exNef associated neuronal damage and leads to greater neurocognitive impairment in the context of cART. In this proposal we will investigate in the context of cART and opiates, the impact of extracellular vesicles, specifically exNef released from HIV-infected (or nef- transfected) microglia on neurons in order to understand the mechanism(s) that underlie HIV-induced neurocognitive impairment/HAND during aviremia. We will also perform a cross-sectional study comparing cerebral spinal fluid(CSF) exNef in PLWHAs on cART with a history/current opiate use and neurocognitive impairment/HAND. Findings from this proposal will allow us to demonstrate the role of EVs in the neuropathogenesis induced by the interplay of opiates and HIV in the CNS.