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

项目摘要 人类免疫缺陷病毒1型（HIV-1）今天影响全世界3 800多万人。虽然目前的 治疗策略有效地抑制了病毒在外周组织、中枢神经系统 仍然容易受到病毒蛋白诱导的损伤，这些蛋白在最初感染后不久进入， 是大多数抗逆转录病毒疗法的靶点。因此，关键是要集中精力发展补充 干预策略，以解决经常出现在感染个体中的持续性炎症， 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（CB 1 R）上调 病毒蛋白质，包括转录反式激活因子（达特），其是与神经毒性最强的病毒蛋白质之一， 患有潜伏的HIV-1感染。内源性大麻素系统是一个很有前途的治疗靶点， 在神经系统损伤模型中减少炎症并恢复神经元功能。事实上，此前 研究表明通过抑制2-花生四烯酸甘油的降解 （2-AG），一种内源性大麻素，用于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。在探索内源性大麻素活性的保护作用时， 对于由达特驱动的毒性的调节，我们先前已经发现通过阻断2-AG的表达上调2-AG， 使用单酰基甘油脂肪酶抑制剂MJN 110的分解在Tat诱导的神经损伤模型中具有神经保护作用 和认知失调。鉴于这些发现，我假设抑制 体内2-AG水解可能下调Tat驱动的神经元兴奋毒性， 行为和减少脑组织中的促炎标志物。然而，达特和MJN 110的作用 尚未使用体内成像来表征。因此，本项目的目标是确定达特和 MJN 110对神经元兴奋性及相应行为、促炎细胞因子表达的影响 IL-6和IL-8，以及大麻素1型受体（CB 1 R）在神经元和小胶质细胞上的密度， 两个具体目标。目标1。在奖励相关的行为任务中使用体内单光子显微镜， 表征在达特和/或MJN 110存在下背内侧前额叶皮质神经元的活动模式。 目标2.利用免疫组织化学标记和多重检测来确定MJN 110是否能够 分别降低细胞特异性CB 1 R密度或促炎细胞因子表达。为此，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， 炎症是潜伏HIV-1感染相关并发症的基础。

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.