Assessing Targeted Cannabinoid Therapeutic Potential Against HIV-1 Associated Neuronal Hyperexcitability and Neuroinflammation

评估针对 HIV-1 相关神经元过度兴奋和神经炎症的靶向大麻素治疗潜力

• 批准号: 10484625
• 负责人: Alexis League
• 金额: $ 3.78万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-15 至 2023-05-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10484625
• 关键词: 2-arachidonylglycerol; Address; Affect; Animals; Anti-Inflammatory Agents; Auditory; Behavior; Behavioral; Brain; CNR1 gene; Cells; Chronic; Cognitive; Cognitive deficits; Confocal Microscopy; Cues; Data; Decision Making; Defense Mechanisms; Development; Disease; Endocannabinoids; Enzymes; Exposure to; Genetic Transcription; Goals; HIV-1; Health; Hydrolysis; IL8 gene; Image; In Vitro; Individual; Infection; Inflammation; Inflammatory; Interleukin-6; Intervention; Label; Longevity; MAGL inhibitor; Measures; Microglia; Microscopy; Modeling; Monoacylglycerol Lipases; Motivation; Nerve Degeneration; Nervous system structure; Neuraxis; Neuronal Dysfunction; Neurons; Outcome; Pattern; Peripheral; Peripheral Nervous System; Persons; Phenotype; Photons; Predisposition; Prefrontal Cortex; Property; Proteins; Regulation; Rewards; Stains; Survival Rate; Therapeutic; Tissues; Toxic effect; Training; Trans-Activators; Treatment Efficacy; Up-Regulation; Viral Load result; Viral Proteins; Virus; Virus Replication; Work; antiretroviral therapy; behavioral outcome; brain tissue; cannabinoid treatment; classical conditioning; conditioning; cytokine; density; endogenous cannabinoid system; excitotoxicity; experimental study; in vivo; in vivo calcium imaging; in vivo imaging; inflammatory marker; multiplex assay; neurocognitive disorder; neuroinflammation; neuronal excitability; neurophysiology; neurotoxic; novel therapeutics; protective effect; receptor density; receptor expression; relating to nervous system; response; therapeutic target; treatment strategy

PROJECT SUMMARY Human immunodeficiency virus type 1 (HIV-1) affects over 38 million people worldwide today. While current treatment strategies effectively suppress virus replication in peripheral tissues, the central nervous system remains vulnerable to damage induced by viral proteins, which enter soon after initial infection and are not well targeted by most antiretroviral therapies. It is thus critical to focus efforts on development of supplemental intervention strategies to address persistent inflammation which presents frequently in infected individuals as neuronal dysregulation and cognitive deficits, especially involving motivation and reward-related decision making. Interestingly, neuronal cannabinoid type-1 receptors (CB1R) are upregulated when exposed to HIV-1 viral proteins including transactivator of transcription (Tat), one of the most neurotoxic viral proteins associated with latent HIV-1 infection. The endogenous cannabinoid system is a promising therapeutic target as activation reduces inflammation and restores neuronal function in models of nervous system insult. Indeed, previous studies have shown reductions in measures of inflammation by inhibiting degradation of 2-arachidonoylglycerol (2-AG), an endocannabinoid produced to promote cellular responses to damage in tissue or neighboring cells and regulate activity of neurons. In exploring the protective effects of endogenous cannabinoid activity modulation against toxicity driven by Tat, we have previously found that upregulating 2-AG through blocking its breakdown using monoacylglycerol lipase inhibitor MJN110 is neuroprotective in models of Tat-induced neural and cognitive dysregulation in vitro and in vivo, respectively. Given these findings, I hypothesize that inhibiting hydrolysis of 2-AG in vivo may downregulate Tat-driven neuronal excitotoxicity during reward-related behavior and reduce proinflammatory markers in brain tissue. However, the effects of Tat and MJN110 have not yet been characterized using in vivo imaging. Thus, the goal of this project is to determine how Tat and MJN110 influence excitability of neurons and corresponding behavior, expression of proinflammatory cytokines IL-6 and IL-8, as well as density of cannabinoid type-1 receptors (CB1R) on neurons and microglia in the following two Specific Aims. Aim 1. To use in vivo one-photon microscopy during a reward-related behavioral task to characterize activity patterns of dorsomedial prefrontal cortex neurons in the presence of Tat and/or MJN110. Aim 2. To use immunohistochemical labeling and multiplex assays to determine whether MJN110 is able to reduce cell-specific CB1R density or proinflammatory cytokine expression, respectively. In doing so, I will enhance our understanding of endocannabinoid therapeutic potential against neuronal dysfunction and inflammation which underlie complications associated with latent HIV-1 infection.

项目概要 目前，1 型人类免疫缺陷病毒 (HIV-1) 影响着全球超过 3800 万人。目前 治疗策略有效抑制病毒在周围组织、中枢神经系统中的复制 仍然容易受到病毒蛋白引起的损害，病毒蛋白在初次感染后不久就会进入，并且状态不佳 大多数抗逆转录病毒疗法的目标。因此，关键是要集中精力开发补充性的 解决感染者中经常出现的持续性炎症的干预策略 神经元失调和认知缺陷，特别是涉及动机和奖励相关决策 制作。有趣的是，当暴露于 HIV-1 时，神经元大麻素 1 型受体 (CB1R) 会上调 病毒蛋白，包括转录反式激活蛋白 (Tat)，是最具神经毒性的病毒蛋白之一 具有潜伏性 HIV-1 感染。内源性大麻素系统是一个有前途的治疗靶点 在神经系统损伤模型中减少炎症并恢复神经元功能。确实，之前的 研究表明，通过抑制 2-花生四烯酰甘油的降解可减少炎症反应 (2-AG)，一种内源性大麻素，可促进细胞对组织或邻近细胞损伤的反应 并调节神经元的活动。探索内源性大麻素活性的保护作用 针对 Tat 驱动的毒性调节，我们之前发现通过阻断其上调 2-AG 使用单酰基甘油脂肪酶抑制剂 MJN110 进行分解在 Tat 诱导的神经模型中具有神经保护作用 以及分别在体外和体内的认知失调。鉴于这些发现，我假设抑制 2-AG 体内水解可能会下调奖赏相关期间 Tat 驱动的神经元兴奋性毒性 行为并减少脑组织中的促炎标记物。然而，Tat 和 MJN110 的效果 尚未使用体内成像进行表征。因此，该项目的目标是确定 Tat 和 MJN110影响神经元的兴奋性和相应的行为、促炎细胞因子的表达 IL-6 和 IL-8，以及以下神经元和小胶质细胞上大麻素 1 型受体 (CB1R) 的密度 两个具体目标。目标 1. 在奖励相关行为任务中使用体内单光子显微镜 表征存在 Tat 和/或 MJN110 时背内侧前额叶皮层神经元的活动模式。 目标 2. 使用免疫组织化学标记和多重检测来确定 MJN110 是否能够 分别降低细胞特异性 CB1R 密度或促炎细胞因子表达。这样做时，我将 增强我们对内源性大麻素治疗神经元功能障碍潜力的理解 炎症是潜在 HIV-1 感染相关并发症的基础。