Methamphetamine Effect on HIV Persistence

项目总结 人类免疫缺陷病毒(HIV)-1感染经常与滥用精神刺激剂有关 毒品，如冰毒(冰毒)。病毒和精神刺激剂的相互作用，特别是在 对于病毒持久性的影响，人们知之甚少，将在这里使用体外和体内方法进行研究。我们 最近观察到，阻断p38 MAPK和敲除或缺失lncRNA linc02574-201可以 抑制HIV-1复制。因此，在本申请中，我们建议研究(S)通过 METH明显促进HIV-1感染以及p38 MAPK和lncRNA linc02574的潜在贡献- 201.提出了三个具体目标：1)调查甲基苯丙胺(冰毒)如何增加HIV-1 外周血淋巴细胞和单核/巨噬细胞感染。这一目标将检验这一假设 冰毒暴露通过促进HIV-1感染，以浓度和时间依赖的方式影响HIV-1感染 P38MAPK活性和lnRNA linc02574-201的表达。2)研究冰毒如何干扰病毒 通过联合抗逆转录病毒治疗(CART)进行抑制。这一目标将调查冰毒暴露如何影响 一旦ARV治疗停止，CART对病毒的抑制和病毒的复发。这一目标也将 评估抑制或敲除p38MAPK是否可以防止CART后病毒复制的恢复，类似 与LncRNA LINC02574-201的击倒有关。3)在人源化CD34-HSC移植的NSG小鼠中进行评估 模型如果冰毒增加了病毒库，并促进了艾滋病毒在CART中断期间的复发。这 AIM将研究冰毒如何影响HIV-1感染、CART对病毒的抑制以及病毒在 在HIV允许的体内模型中阻断CART，CD34造血干细胞(HSC)植入 NSG小鼠模型。这些实验将在体外用分离的人类外周血细胞和 细胞系，以及体内人源化小鼠的CD34细胞来源的造血系统提供HIV 人外周CD4T淋巴细胞和巨噬细胞(MΦ)。所有三个具体目标都将定义 HIV1感染的CD4T细胞和MΦ在冰毒存在和不存在情况下的核糖核酸特征 测序，并将测试所提出的机制，包括增加应激相关的p38MAPK的活性 LncRNA linc02574-201表达上调。AIMS还将监测已知的支持HIV-1感染的因素， 例如IRF7，并将测试冰毒暴露下调包括以下ISG子集的前提 抗病毒因子，如CCL5、Mx1/2、IFITM2/3和IRF1和-3。

PROJECT SUMMARY Infection with Human Immunodeficiency virus (HIV)-1 is frequently associated with abuse of psychostimulant drugs, such as methamphetamine (METH). The interaction of virus and psychostimulant, in particular with regard to viral persistence, is poorly understood and will be studied here using in vitro and in vivo approaches. We recently observed that blockade of p38 MAPK and knockdown or deletion of the lncRNA linc02574-201 can inhibit HIV-1 replication. Therefore, we propose in this application to investigate the mechanism(s) by which METH apparently promotes HIV-1 infection and the potential contributions of p38 MAPK and lncRNA linc02574- 201. Three Specific Aims are proposed: 1) To investigate how methamphetamine (METH) increases HIV-1 infection of peripheral blood lymphocytes and monocytes/macrophages. This aim will test the hypothesis that METH exposure affects HIV-1 infection in a concentration- and timing-dependent fashion by promoting the activity of p38 MAPK and expression of lnRNA linc02574-201. 2) To study how METH interferes with viral suppression by combined antiretroviral therapy (cART). This aim will investigate how METH exposure affects viral suppression by cART and the recrudescence of the virus once ARV treatment ceases. This aims will also assess if inhibition or knockdown of p38 MAPK can prevent the resumption of viral replication after cART, similar to the knockdown of lncRNA linc02574-201. 3) To assess in a humanized CD34+ HSC-engrafted NSG mouse model if METH increases the viral reservoir and facilitates the recurrence of HIV during interruption of cART. This aim will study how METH affects HIV-1 infection, viral suppression by cART and viral recrudescence upon interruption of cART in an HIV permissive in vivo model, the CD34+ HSC (hematopoietic stem cell)-engrafted NSG mouse model. The experiments will be performed in vitro with isolated human peripheral blood cells and cell lines, and in vivo in humanized mice with a CD34+ cell-derived hematopoietic system that provides HIV permissive human peripheral CD4+ T-lymphocytes and macrophages (MΦ). All three Specific Aims will define RNA signatures of HIV-1 infected CD4+ T-cells and MΦ in the presence and absence of METH using RNA- sequencing and will test the proposed mechanism, including increased activity of the stress-related p38 MAPK and up-regulation of lncRNA linc02574-201. The aims will also monitor factors known to support HIV-1 infection, such as IRF7, and will test the premise that METH exposure down-regulates a subset of ISGs that include antiviral factors, such as CCL5, Mx1/2, IFITM2/3 and IRF1 and -3.