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

大麻二酚对慢性感染免疫功能小鼠的 HIV 表达、神经炎症和 HIV 认知疾病的影响机制

• 批准号: 10686313
• 负责人: Alejandra Borjabad
• 金额: $ 70.55万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-30 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10686313
• 关键词: Affect; Anti-Inflammatory Agents; Anti-Retroviral Agents; Antigens; Antiinflammatory Effect; Anxiety; Apoptosis; Archives; Astrocytes; Basic Science; Biochemical; Bioinformatics; Biological Assay; Brain; Brain Diseases; CB2 knockout; CNR1 gene; CNR2 gene; Cannabidiol; Cannabinoids; Cannabis; Cell Lineage; Cell Separation; Cells; Chronic; Cognition Disorders; Critical Pathways; DNA; Detection; Disease; Dose; Epigenetic Process; Evaluation; Flow Cytometry; Frequencies; Gene Expression; Gene Expression Profiling; Genes; HIV; HIV Infections; HIV Seropositivity; HIV-associated neurocognitive disorder; Hand; Hypermethylation; IL7 gene; Immune; Immune response; Immune system; Immunity; Immunocompetent; Immunologics; Immunosuppression; Impaired cognition; Impairment; In Vitro; Individual; Infection; Inflammation; Inflammatory; Innate Immune Response; Interferon Receptor; Interferons; Joints; Knockout Mice; Learning; Lymphocyte; Macrophage; Measurement; Mediating; Medicine; MicroRNAs; Microglia; Modeling; Modification; Mononuclear; Mus; National NeuroAids Tissue Consortium; Neurocognitive Deficit; Neuropathy; Pain; Pathology; Pathway interactions; Patients; Peripheral; Peripheral Nervous System Diseases; Persons; Pharmaceutical Preparations; Phosphorylation; Poly I-C; Population; Research; Residual state; Role; Route; Science; Seminal fluid; Signal Pathway; Specificity; Stimulus; System; Systems Biology; Testing; Time; Tissues; Viral; Virus; adaptive immunity; antagonist; antiretroviral therapy; antiviral immunity; behavior test; brain cell; brain dysfunction; brain tissue; chronic infection; drug of abuse; experience; high risk; histone modification; in vivo; inflammatory marker; marijuana use; marijuana user; neuroinflammation; neuropathology; pain relief; posttranscriptional; pre-clinical; pre-clinical research; programs; promoter; receptor; response; single-cell RNA sequencing; transcriptome; transcriptome sequencing; transmission process

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

本申请是根据RFA-DA-20-022提交的，并提出了基本要求 研究 非精神活性物质 PLWH通常会经历这种情况。然而，其长期的HIV感染和HIV的进展 疾病在很大程度上仍未确定。我们对感染嵌合HIV，EcoHIV的小鼠的初步结果表明， CBD在大脑中同时具有抗炎和免疫抑制功能；令人惊讶的是， 感染CBD的小鼠脑内天然抗病毒效应物干扰素-减少。直接 与这种RFA相关的是，我们发现CBD使小鼠大脑中HIV的表达增加了10倍。此外， 尽管CBD单独具有抗炎作用，但该药物未能缓解HIV介导的炎症基因 表情。这些结果表明，使用CBD可以增加PLWH中HIV的表达，从而 加剧艾滋病毒相关疾病，包括非传染性非传染性疾病。我们假设CBD会增加HIV脑部感染 其结果是其抗病毒免疫功能受损，包括干扰素功能。具体目标是：1) 优化CBD介导的EcoHIV脑感染增加及其对小鼠行为的影响 测试包括在抗逆转录病毒药物存在时和在慢性感染期间的反应，CB测试 利用基因敲除小鼠的受体参与，以及从死者脑中进行艾滋病毒负担/表达的试点研究 PLWH使用大麻。2)通过免疫来确定CBD对艾滋病毒控制的有利或有害影响 EcoHIV感染小鼠脑组织中抗原特异性和体内固有反应的检测 以及利用单细胞RNA技术对脑内淋巴细胞、巨噬细胞和小胶质细胞的基因表达谱进行研究。 SEQ和生物信息学。3)使用目标2中的信息，检验CBD会增加HIV的假设 干扰干扰素反应包括转录后分析在脑和NCI中的表达 干扰素诱导修饰、干扰素刺激基因启动子的表观遗传修饰和干扰素特异性 使用基因敲除的小鼠。我们的方法将包括CBD和EcoHIV的抗逆转录病毒联合治疗- 感染小鼠，QPCR测量组织中的病毒载量，学习，隔离和 来自大脑的单个核细胞的功能评估，脑细胞谱系特异性RNA序列，机制 CBD对干扰素信号通路的影响及决定干扰素反应基因的表观遗传学变化 表达，关键受体敲除小鼠对EcoHIV/CBD的反应，以及来自NNTC的精选脑组织 艾滋病毒感染者使用大麻和不滥用其他药物。此应用程序符合RFA “抗逆转录病毒治疗条件下慢性/持续性艾滋病毒引起炎症的研究模型”的要求； 评价“大麻素在艾滋病毒相关慢性炎症中的作用”；以及“描绘…。。有益的 或使用大麻素对艾滋病毒引起的炎症的有害影响“。 科学与基础研究 在生态艾滋病毒感染的小鼠中模拟大麻二醇(CBD)对艾滋病毒携带者(PLWH)的影响。CBD，a 大麻成分，免费获得，广泛用于缓解疼痛和焦虑 对……的影响