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: 10265583
• 负责人: Alejandra Borjabad
• 金额: $ 72.02万
• 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/10265583
• 关键词: Affect; Anti-Inflammatory Agents; Anti-Retroviral Agents; Antigens; Antiinflammatory Effect; Antiviral Agents; Anxiety; Apoptosis; Archives; Astrocytes; Basic Science; Biochemical; Bioinformatics; Biological Assay; Brain; Brain Diseases; CB2 knockout; CNR1 gene; CNR2 gene; Cannabidiol; Cannabinoids; Cannabis; Cell Lineage; Cells; Chronic; Cognition Disorders; Critical Pathways; DNA; Detection; Disease; Dose; Epigenetic Process; Evaluation; Flow Cytometry; Frequencies; Gene Expression; Gene Expression Profiling; Genes; Genetic Transcription; HIV; HIV Infections; HIV Seropositivity; Hypermethylation; IL7 gene; Immune; Immune response; Immune system; Immunity; Immunocompetent; Immunologics; Impaired cognition; Impairment; In Vitro; Individual; Infection; Inflammation; Inflammatory; Innate Immune Response; Interferons; Joints; Knockout Mice; Learning; Lymphocyte; Measurement; Mediating; MicroRNAs; Microglia; Modeling; Modification; Mononuclear; Mus; National NeuroAids Tissue Consortium; Neurocognitive Deficit; Neuropathy; Pain; Pain-Free; 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; Virus; adaptive immunity; antiretroviral therapy; antiviral immunity; behavior test; brain cell; brain tissue; chronic infection; drug of abuse; experience; gene function; high risk; histone modification; in vivo; inflammatory marker; macrophage; marijuana use; marijuana user; neuroinflammation; pain relief; pre-clinical; pre-clinical research; programs; promoter; receptor; response; single-cell RNA sequencing; transcriptome; transcriptome sequencing; transmission process; virology

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

本申请是为了响应 RFA-DA-20-022 而提交的，并提出了基本的建议 研究 非精神活性的 PLWH 普遍经历的情况。然而，其长期的艾滋病毒感染和艾滋病毒的进展 疾病在很大程度上仍不确定。我们对感染嵌合 HIV、EcoHIV 的小鼠的初步结果显示 CBD 在大脑中具有抗炎和免疫抑制功能；引人注目的规范 在受 CBD 治疗的受感染小鼠的大脑中，先天抗病毒效应物干扰素 (IFN)- 减少。直接地 与此 RFA 相关的是，我们发现 CBD 可使小鼠大脑中的 HIV 表达增加多达 10 倍。而且， 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 表达。这些结果表明，CBD 的使用可以增加 PLWH 中的 HIV 表达，从而 加剧包括 NCI 在内的 HIV 相关疾病。我们假设 CBD 增加了 HIV 脑部感染 这是由于其抗病毒免疫功能（包括干扰素功能）受损所致。具体目标是：1) 优化 CBD 介导的 EcoHIV 脑部感染增加并分析其对小鼠行为的影响 测试包括抗逆转录病毒药物存在下和慢性感染期间的反应、CB测试 使用基因敲除小鼠进行受体参与，并对死者大脑进行艾滋病毒负荷/表达试点研究 使用大麻的艾滋病毒感染者。 2) 确定 CBD 对免疫控制 HIV 的有益或有害影响 EcoHIV 感染小鼠的大脑，包括体外抗原特异性反应、体内先天反应的测定 通过单细胞 RNA 进行脑淋巴细胞、巨噬细胞和小胶质细胞的离体和基因表达谱分析 seq 和生物信息学。 3) 使用目标 2 中的信息，检验 CBD 增加 HIV 的假设 通过破坏 IFN 反应（包括转录后测定）在大脑和 NCI 中表达 干扰素诱导的修饰、干扰素刺激基因启动子的表观遗传修饰以及干扰素特异性 使用基因敲除小鼠。我们的方法将包括联合 CBD 和抗逆转录病毒治疗 EcoHIV- 受感染小鼠、用于测量组织中病毒负荷的 QPCR、学习、隔离和感染的行为测试 大脑单核细胞的功能评估、脑细胞谱系特异性 RNA seq、机制 CBD 对 IFN 信号通路影响的研究以及决定 IFN 反应基因的表观遗传变化 表达、对 EcoHIV/CBD 做出反应的关键受体敲除小鼠以及来自 NNTC 的精选脑组织 艾滋病毒感染者使用大麻而不滥用其他药物。此应用程序符合 RFA “ART条件下慢性/持续性HIV诱发炎症的研究模型”的要求；为了 评估“大麻素在艾滋病毒相关慢性炎症中的作用”；并“描绘……有益的 或大麻素的使用对艾滋病毒引起的炎症的有害影响”。 科学和临床前 在感染 EcoHIV 的小鼠中模拟大麻二酚 (CBD) 对艾滋病毒感染者 (PLWH) 的影响。 CBD，一个 大麻成分，可免费获得并广泛用于缓解疼痛和焦虑 影响于