The impact of methamphetamine on CXCL12 mediated HIV neuropathogenesis

这项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CD14+CD16+单核细胞通过血脑屏障迁移到CXCL12，增加血管周围和 感染实质的巨噬细胞在HIV(PWH)合并冰毒使用障碍患者的中枢神经系统聚集。 HIV相关神经认知障碍或损害(手，HIV-NCI)的患病率在 即使接受抗逆转录病毒治疗(ART)，PWH也会极大地影响他们的生活质量。相当数量的 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。 在使用PWH的活性冰毒中。抗逆转录病毒治疗中冰毒使用障碍增加PWH患者HIV-NCI的机制 没有完全的特征，阻碍了减少或消除干预策略的发展 这一人群中的认知功能障碍。HIV在最初感染后早期进入中枢神经系统，部分是通过趋化因子 感染和未感染的CD14+CD16+单核细胞通过血脑屏障的诱导迁移 (BBB)，有助于病毒储备库的补充和慢性低水平神经炎， 描述HIV-NCI的特征。趋化因子CXCL12(SDF-1)在中枢神经系统低水平结构性表达 并且在PWH的中枢神经系统中增加，提示这种趋化因子有助于未感染和 感染CD14+CD16+单核细胞进入中枢神经系统。我们实验室报告了一项新发现，CXCR7或 ACKR3是CXCL12的非典型趋化因子受体，表达于未感染和HIV的表面 感染CD14+CD16+单核细胞，并与CXCR4一起促进这些细胞的迁移 穿过BBB到达CXCL12。我们的初步数据表明，冰毒增加了CXCL12诱导的转位 未感染和HIV感染的CD14+CD16+单核细胞。CXCR7和/或CXCR4在这些冰毒中作用 介导效应，特别是在携带艾滋病毒(HIV+)的CD14+CD16+单核细胞迁移增加方面 与暴露于病毒(HIVexp)但不携带病毒的细胞相比，HIVexp是这项提议的重点。我们 假设CXCR7和/或CXCR4参与了甲基介导的CXCL12诱导的HIV感染增加 感染的CD14+CD16+单核细胞迁移，包括HIV+细胞的优先迁移 与HIVexp细胞相比。因此，CXCR7可能是甲基苯丙胺治疗HIV-NCI的治疗靶点 使用无序。我们将使用我们的体外人血脑屏障模型来表征CXCR7和/或CXCR4在 冰毒对血脑屏障移行和细胞的影响。我们将确定冰毒对CXCR7的影响 和/或CXCR4的表达，以及CXCL12诱导的信号、黏附、趋化和侵袭 未感染/HIV感染的CD14+CD16+单核细胞。冰毒对人体内单核细胞内流的影响 将在经冰毒处理的SIV感染猕猴的中枢神经系统组织切片中检测受感染的CNS CXCL12及其血管周围和实质BBB表达的免疫组织化学分析 表达CXCR7和/或CXCR4的单核/巨噬细胞聚集。

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.