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

项目总结 人类免疫缺陷病毒1型(HIV-1)目前影响着全世界3800万人。当当前 治疗策略有效地抑制了病毒在周围组织、中枢神经系统的复制 仍然容易受到病毒蛋白的破坏，病毒蛋白在最初感染后不久进入，身体状况不佳 是大多数抗逆转录病毒治疗的靶点。因此，将重点放在发展补充技术上是至关重要的 应对持续性炎症的干预策略，这种炎症在感染者中经常出现为 神经元调节失调和认知缺陷，特别是涉及动机和奖励相关的决定 制作。有趣的是，暴露在HIV-1中的神经元大麻素1型受体(CB1R)表达上调 病毒蛋白包括转录反式激活因子(TAT)，这是最具神经毒性的病毒蛋白之一 有潜伏的HIV-1感染。内源性大麻素系统是一种很有前途的激活治疗靶点 减少炎症，恢复神经系统损伤模型的神经功能。事实上，以前的 研究表明，通过抑制2-花生四烯基甘油的降解，可以减少炎症的程度。 (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。探索内源性大麻素活性的保护作用 针对TAT驱动的毒性调制，我们先前发现通过阻断其受体上调2-AG 单甘油脂肪酶抑制剂MJN110对TAT诱导的神经损伤模型的神经保护作用 和认知失调分别在体外和体内。考虑到这些发现，我假设抑制 体内2-AG的水解可能下调奖赏相关过程中TAT驱动的神经元兴奋性毒性 并减少脑组织中的促炎标志物。然而，TAT和MJN110的影响 还没有用活体成像来表征。因此，该项目的目标是确定TAT和 MJN110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，以及神经元和小胶质细胞上大麻素1型受体(CB1R)的密度 有两个明确的目标。目的1.在奖赏相关的行为任务中使用活体单光子显微镜 描述在TAT和/或MJN110存在下背内侧前额叶皮质神经元的活动模式。 目的2.应用免疫组织化学标记和多重分析方法确定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。通过这样做，我将 增强我们对内源性大麻素治疗神经元功能障碍的潜力的理解 与潜伏的HIV-1感染相关的并发症的基础炎症。

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.