Mechanism of Cannabidiol Effects on HIV Expression, Neuroinflammation, and HIV Cognitive Disease in Chronically-infected Immunocompetent Mice

本申请是根据RFA-DA-20-022提交的，并建议基本 研究 非精神活性 通常由PLWH经历。但是，其长期感染艾滋病病毒和艾滋病病毒的进展 疾病在很大程度上仍未确定。我们在感染嵌合HIV（EcoHIV）的小鼠中的初步结果显示， CBD在大脑中具有抗炎和免疫抑制功能;引人注目的是， 先天性抗病毒效应物干扰素（IFN）-γ在受感染的CBD处理的小鼠的脑中减少。直接 与RFA相关，我们表明CBD使小鼠大脑中的HIV表达增加了10倍。此外，委员会认为， 尽管CBD单独具有抗炎作用，但该药物未能减轻HIV介导的炎症基因， 表情这些结果表明，CBD的使用可以增加艾滋病毒在PLWH中的表达， 加剧艾滋病毒相关疾病，包括NCI。我们假设CBD引起的HIV脑感染增加 结果是其损害了包括IFN功能在内的抗病毒免疫。具体目标是：1） 优化CBD介导的EcoHIV脑感染增加，并测定其对小鼠行为的影响 测试，包括在抗逆转录病毒药物的存在和慢性感染期间的反应，CB测试 使用基因敲除小鼠进行的受体参与研究，以及对死者大脑进行的HIV负荷/表达试验研究。 使用大麻的艾滋病毒携带者。2)确定CBD对艾滋病毒控制的有益或有害影响，通过免疫， EcoHIV感染小鼠的脑，包括离体抗原特异性应答、体内先天性应答的测定 和离体，和脑淋巴细胞，巨噬细胞和小胶质细胞的基因表达谱通过单细胞RNA- seq和生物信息学。3)使用目标2中的信息，测试CBD增加HIV的假设 通过干扰素应答的破坏在脑和NCI中的表达，包括转录后表达的测定。 IFN诱导中的修饰、IFN刺激基因启动子中的表观遗传修饰和IFN特异性 使用基因敲除小鼠。我们的方法将包括CBD和EcoHIV的抗逆转录病毒治疗相结合， 感染小鼠，用于测量组织中病毒负荷的QPCR，学习、隔离和 来自脑的单核细胞的功能评价，脑细胞谱系特异性RNA测序，机制 CBD对IFN信号通路的影响及决定IFN应答基因的表观遗传学变化的研究 表达，关键受体敲除小鼠对EcoHIV/CBD的反应，以及来自NNTC的 使用大麻而不滥用其他药物的艾滋病毒感染者。此应用程序符合RFA “ART条件下慢性/持续性HIV诱导炎症的研究模型”的要求; 评估“大麻素在艾滋病毒相关慢性炎症中的作用”;以及“描述.....有益 或大麻素使用对艾滋病毒引起的炎症的有害影响”。 科学和临床前 在EcoHIV感染的小鼠中模拟大麻二酚（CBD）对HIV感染者（PLWH）的影响。生物多样性公约a 大麻成分，是免费提供和广泛用于缓解疼痛和焦虑 影响

This application is submitted in response to RFA-DA-20-022 and proposes basic research non-psychoactive commonly experienced by PLWH. However, its long-term HIV infection and progression of HIV disease remain largely undetermined. Our preliminary results in mice infected by chimeric HIV, EcoHIV, show that CBD has both anti-inflammatory and immunosuppressive functions in the brain; strikingly the canonical innate antiviral effector, interferon (IFN)-, was reduced in brains of infected CBD treated mice. Directly relevant to this RFA, we show that CBD increases HIV expression in mouse brain up to 10-fold. Moreover, despite anti-inflammatory effects of CBD alone, the drug failed to mitigate HIV mediated inflammatory gene expression. These results suggest that CBD use can increase HIV expression in PLWH and thereby exacerbate HIV-related diseases including NCI. We hypothesize that increased HIV brain infection by CBD results from its impairment of antiviral immunity including IFN functions. The Specific Aims are to: 1) Optimize CBD mediated increase in EcoHIV brain infection and assay its consequences in mouse behavioral tests including responses in the presence of antiretroviral drugs and during chronic infection, tests of CB receptor participation using knockout mice, and pilot HIV burden/ expression studies from brains of deceased PLWH using cannabis. 2) Determine beneficial or deleterious effects of CBD on HIV control by immunity in brains of EcoHIV-infected mice including assay of antigen-specific responses ex vivo, innate responses in vivo and ex vivo, and gene expression profiling of brain lymphocyte, macrophage and microglia by single cell RNA- seq and bioinformatics. 3) Using information from Aim 2, test the hypothesis that CBD increases HIV expression in the brain and NCI by disruption of IFN responses including assay of post-transcriptional modification in IFN induction, epigenetic modification in promoters of IFN stimulated genes, and IFN specificity using knockout mice. Our approaches will include combined CBD and antiretroviral treatment of EcoHIV- infected mice, QPCR for measurement of virus burden in tissues, behavioral tests of learning, isolation and functional evaluation of mononuclear cells from the brain, brain cell lineage specific RNA seq, mechanistic studies of CBD effects upon IFN signaling pathways and epigenetic changes determining IFN responsive gene expression, key receptor knockout mice for responses to EcoHIV/CBD, and curated brain tissue from NNTC of HIV-infected persons using cannabis and no other drugs of abuse. This application meets the RFA requirements of “research models of chronic/persistent HIV-induced inflammation under conditions of ART”; for evaluation of “the role of cannabinoids in HIV-associated chronic inflammation”; and to “Delineate ….. beneficial or deleterious effects of cannabinoid use on HIV-induced inflammation”. science and preclinical in EcoHIV-infected mice to model cannabidiol (CBD) effects in people living with HIV (PLWH). CBD, a cannabis component, is freely available and widely used for relieving pain and anxiety effects on