Defining the impact of cannabinoids on the HIV reservoir in humanized mice

摘要 大麻(CB)的使用在艾滋病毒携带者(PWH)中很普遍，但它对艾滋病毒感染和潜伏的影响 尚未对病毒库进行全面检查。特别是，CB对固体组织中HIV储存库的影响 比如大脑是完全未知的。因此，可以仔细控制给药的动物模型 而组织样本的分析对于全面了解CB如何影响HIV宿主将是重要的 组织中的特征。我们的中心假设是CB诱导了一种免疫抑制的细胞状态， 影响病毒转录和组织中艾滋病毒储存库的维持。这项提案的总体目标是 建立在HIV感染期间接触CB的人源化小鼠模型，并使用该模型学习如何 CB的使用会影响组织内HIV储存库的特性。在R61阶段，艾滋病毒感染人性化 表达hIL-34促进小胶质细胞中枢神经系统重建的小鼠将暴露于不同剂量的THC 在感染期间，在使用抗逆转录病毒治疗(ART)抑制病毒之前和之后。货柜码头处理费的影响 将检查外周血中病毒血症、病毒抑制、免疫标志物和细胞因子的暴露情况。 在R33阶段，我们将使用此模型来解决三个主要问题。第一，病毒DNA(VDNA)和病毒 感染了THC的小鼠大脑和脾中的RNA(VRNA)水平将 量化以确定THC对这些病毒储存库的大小和转录活性的影响 纸巾。其次，我们将检查暴露于THC是否会影响体内HIV潜伏期的逆转。我们假设 由有效的潜伏期反转剂(LRA)激活的炎症信号通路之间的串扰 如非规范的核因子-kB激动剂AZD5582和使用CB激活的抗炎通路可能 显著改变LRA体内给药的结果。为了验证这一假设，THC或对照暴露的艾滋病毒 感染的小鼠将被给予AZD5582，以及THC对组织中病毒血症和病毒重新激活的影响 将会被检查。最后，我们将使用该模型对受感染的细胞进行深入的单细胞表征 在存在和不存在THC暴露的情况下，在ART抑制的人源化小鼠的大脑和脾中。 脾和脑组织将使用联合单细胞ATACseq/RNAseq方法进行分析，以识别和 鉴定HIV vDNA或vRNA细胞，并比较这些细胞的丰度和表型 CB给小鼠和对照组灌胃。如果成功，这项提案将揭示CB如何影响艾滋病毒的新见解 感染和组织驻留艾滋病毒储存库的特点。这反过来又将指导新的艾滋病毒治疗方法 针对CB-使用PWH进行了优化的策略。

Abstract Cannabis (CB) use is prevalent amongst people with HIV (PWH) but its impact on HIV infection and the latent viral reservoir has not been fully examined. In particular, the impact of CB on the HIV reservoir in solid tissues such as the brain is completely unknown. As such, animal models in which dosing can be carefully controlled and tissue samples can be analyzed will be important to fully understand how CB impacts HIV reservoir characteristics in tissues. Our central hypothesis is that CB induces an immunosuppressive cellular state that impacts viral transcription and maintenance of the HIV reservoir in tissues. The overall goal of this proposal is to develop a humanized mouse model of CB exposure during HIV infection and to use this model to learn how CB use affects characteristics of the tissue resident HIV reservoir. In the R61 phase, HIV infected humanized mice expressing hIL-34 to promote microglial CNS reconstitution will be exposed to a range of THC doses during infection, both before and after viral suppression with antiretroviral therapy (ART). The impact of THC exposure on viremia, viral suppression, immune markers and cytokines in peripheral blood will be examined. In the R33 phase, we will use this model to address three major questions. First, viral DNA (vDNA) and viral RNA (vRNA) levels in the brains and spleens of infected mice that have been dosed with THC will be quantified to determine the impact of THC on the size and transcriptional activity of the viral reservoir in these tissues. Second, we will examine whether THC exposure affects HIV latency reversal in vivo. We hypothesize that cross-talk between inflammatory signaling pathways activated by potent latency reversing agents (LRAs) such as the non-canonical NF-kB agonist AZD5582 and anti-inflammatory pathways activated by CB use could significantly alter the outcome of LRA dosing in vivo. To test this hypothesis, THC or control exposed HIV infected mice will be dosed with AZD5582, and the impact of THC on viremia and viral reactivation in tissues will be examined. Finally, we will use this model to perform a deep single cell characterization of infected cells in the brains and spleens of ART-suppressed humanized mice in the presence and absence of THC exposure. Spleen and brain tissues will be profiled using a combined single cell ATACseq/RNAseq method to identify and characterize HIV vDNA+ or vRNA+ cells and to compare the abundance and phenotype of these cells between CB dosed mice and controls. If successful, this proposal will reveal new insights into how CB affects HIV infection and the characteristics of the tissue-resident HIV reservoir. This in turn will guide novel HIV cure strategies that are optimized for CB-using PWH.