The impact of methamphetamine on CXCL12 mediated HIV neuropathogenesis

该提案的目标是描述甲基苯丙胺（冰毒）增加艾滋病毒的机制 感染的CD 14 + CD 16+单核细胞穿过BBB迁移到CXCL 12，增加血管周围和 实质感染的巨噬细胞在患有冰毒使用障碍的HIV（PWH）患者的CNS中积聚。 艾滋病毒相关的神经认知障碍或损害（HAND，HIV-NCI）的患病率增加， PWH，即使使用抗逆转录病毒治疗（ART），也会极大地影响他们的生活质量。相当多的 患有冰毒使用障碍的人也感染了艾滋病毒，据报道，神经认知障碍增加 使用PWH的活跃冰毒甲基苯丙胺使用障碍增加接受抗逆转录病毒治疗的PWH中HIV-NCI的机制 没有完全定性，阻碍了制定干预战略，以减少或消除 认知功能障碍HIV在初次感染后早期进入CNS，部分是通过趋化因子 诱导感染和未感染的CD 14 + CD 16+单核细胞穿过血脑屏障 (BBB)，有助于补充病毒库和慢性低水平神经炎症， 表征HIV-NCI。趋化因子CXCL 12（SDF-1）在CNS中以低水平组成型表达 并且在PWH的CNS中增加，这表明这种趋化因子有助于未感染的和 感染的CD 14 + CD 16+单核细胞进入CNS。我们的实验室报告了CXCR 7或 ACKR 3是CXCL 12的非典型趋化因子受体，在未感染和HIV感染的细胞表面表达。 感染的CD 14 + CD 16+单核细胞，并与CXCR 4一起沿着促进这些细胞的迁移 穿过BBB到达CXCL 12。我们的初步数据表明，甲基增加CXCL 12诱导的迁移 未感染和HIV感染的CD 14 + CD 16+单核细胞。CXCR 7和/或CXCR 4在这些方法中的作用 介导的作用，特别是携带HIV（HIV+）的CD 14 + CD 16+单核细胞的迁移增加 与暴露于但不携带病毒（HIVexp）的细胞相比，这是该提议的重点。我们 假设CXCR 7和/或CXCR 4有助于甲氧西林介导CXCL 12诱导的HIV增加 受感染的CD 14 + CD 16+单核细胞迁移，包括HIV+细胞的优先迁移 与HIVexp细胞相比。因此，CXCR 7可能是PWH中HIV-NCI治疗的治疗靶点， 使用障碍。我们将使用我们的体外人血脑屏障模型来表征CXCR 7和/或CXCR 4在 甲基介导的对迁移和对BBB细胞的影响。我们将确定冰毒对CXCR 7的影响 和/或CXCR 4表达，以及CXCL 12诱导的信号传导，粘附，趋化性和侵袭。 未感染/HIV感染的CD 14 + CD 16+单核细胞。甲氨蝶呤对单核细胞内流至 感染的CNS将在来自甲氨蝶呤处理的SIV感染的猕猴的CNS组织切片中进行检查， BBB中CXCL 12表达的免疫组织化学分析，以及血管周围和实质 CXCR 7和/或CXCR 4表达单核细胞/巨噬细胞的积聚。

The goal of this proposal is to characterize mechanisms by which methamphetamine (meth) increases HIV infected CD14+CD16+ monocyte transmigration across the BBB to CXCL12, increasing perivascular and parenchymal infected macrophage accumulation in the CNS of people with HIV (PWH) with meth use disorder. There is increased prevalence of HIV associated neurocognitive disorders or impairment (HAND, HIV-NCI) in PWH, even with antiretroviral therapy (ART), greatly impacting their quality of life. A significant number of people with meth use disorder are also infected with HIV, with increased neurocognitive impairments reported in active meth using PWH. The mechanisms by which meth use disorder increases HIV-NCI in PWH on ART are not completely characterized, impeding the development of interventional strategies to reduce or eliminate cognitive dysfunctions in this population. HIV enters the CNS early after initial infection, in part, by chemokine induced transmigration of infected and uninfected CD14+CD16+ monocytes across the blood brain barrier (BBB), contributing to the replenishment of viral reservoirs and chronic low level neuroinflammation that characterize HIV-NCI. The chemokine CXCL12 (SDF-1) is constitutively expressed at low levels in the CNS and is increased in the CNS of PWH, suggesting that this chemokine contributes to influx of uninfected and infected CD14+CD16+ monocytes into the CNS. Our laboratory reported the novel finding that CXCR7 or ACKR3, an atypical chemokine receptor for CXCL12, is expressed on the surface of uninfected and HIV infected CD14+CD16+ monocytes and contributes, along with CXCR4, to the transmigration of these cells across the BBB to CXCL12. Our preliminary data indicate that meth increases CXCL12 induced transmigration of both uninfected and HIV infected CD14+CD16+ monocytes. The role of CXCR7 and/or CXCR4 in these meth mediated effects, particularly in increased transmigration of CD14+CD16+ monocytes that harbor HIV (HIV+) compared to cells that are exposed to, but do not harbor, virus (HIVexp), is the focus of this proposal. We hypothesize that CXCR7 and/or CXCR4 contribute to meth mediated increases in CXCL12 induced HIV infected CD14+CD16+ monocyte transmigration, including the preferential transmigration of HIV+ cells compared to HIVexp cells. Thus, CXCR7 may be a therapeutic target for HIV-NCI treatment in PWH with meth use disorder. We will use our in vitro human BBB model to characterize the role of CXCR7 and/or CXCR4 in meth mediated effects on transmigration and on cells of the BBB. We will determine effects of meth on CXCR7 and/or CXCR4 expression, and on CXCL12 induced signaling, adhesion, chemotaxis, and invasion in uninfected/HIV infected CD14+CD16+ monocytes. The in vivo effects of meth on monocyte influx into the infected CNS will be examined in CNS tissue sections from meth treated SIV infected macaques by immunohistochemical analyses of BBB expression of CXCL12, and perivascular and parenchymal accumulation of CXCR7 and/or CXCR4 expressing monocytes/macrophages.