The role of cannabinoids in the regulation of the blood brain barrier in the context of NeuroHIV and anti-retroviral therapy

项目摘要/摘要 尽管接受了抗逆转录病毒疗法(Art)，但神经认知并发症在人们中仍然非常普遍。 艾滋病毒携带者(PLWH)。一种解释可能是血脑屏障(BBB)的不断妥协 由慢性炎症反应驱动。将医用大麻引入HIV治疗实践 似乎对几种病毒相关的并发症(从慢性疼痛到食欲)都有好处 刺激)。然而，大麻对艾滋病毒相关慢性炎症的影响和机制， 对内源性大麻素系统、免疫调节和神经功能障碍的了解甚少。如图所示 在RFA中，临床前模型可以提供对分子和细胞的严格的深入分析 植物大麻素、艾滋病毒和抗逆转录病毒药物之间交叉的机制。为此，我们提出一项 两种最常用的大麻类化合物(THC、CBD)对血脑屏障功能的综合评价 免疫-内皮相互作用和神经炎症。我们将利用最先进的芯片微流控技术 神经血管单位模型(NVU)和艾滋病毒动物模型(w/w/o ART)。之前，我们发现 脑内皮细胞上调HIV感染人脑组织中的CB2。我们还发现， CB2的调节影响HIV的病理指标(体内)，并调节血脑屏障。我们的初步研究 确定植物大麻素可能对血脑屏障的不同性质产生的不同影响。具体来说， 大麻素(THC，CBD)本身就可以增强物理屏障，部分降低内皮细胞的激活和 增强外排转运蛋白的活性。尽管这些影响中的一些似乎是有益的，但艾滋病毒的存在 ART改变了血脑屏障的功能受大麻类物质调节的方式。例如， THC增强的转运蛋白活性对于改变ART-CNS的穿透性有重要的考虑。因此， 我们假设植物大麻素不同地调节血脑屏障功能，这两种功能都是有益的。 对神经艾滋病毒是有害的。在目标1中，使用我们最新的组织工程微流体NVU模型，我们将 分析血脑屏障通透性、转运体状态和免疫内皮细胞的动态变化 互动。然后，在目标2中，我们将比较广泛使用的大麻素途径的结果 使用两种相关的艾滋病毒感染模型(人源化小鼠和吸入型小鼠)体内给药(口服与吸入) 无菌性脑膜炎/脑炎模型)。实验将在ART的背景下评估血脑屏障的变化 和大麻素的接触。最后，我们建议确定新的串扰机制，桥接大麻素 受体信号转导控制血脑屏障维持的信号(目标3)。很明显，大麻类物质对人体产生未知的 细胞特异性效应，有助于对这些化合物如何影响神经艾滋病毒进行混乱的解释。 使用创新的临床前工具，我们的研究将大大有助于理解 在现代神经艾滋病毒时代，大麻类药物使用对血脑屏障的影响。

PROJECT SUMARY/ABSTRACT Despite antiretroviral therapy (ART), neurocognitive complications continue to be highly prevalent in people living with HIV (PLWH). One explanation could be the constant compromise of the blood brain barrier (BBB) driven by chronic inflammatory responses. The introduction of medicinal marijuana into HIV treatment practice appear to be beneficial for several virus associated complications (ranging from chronic pain to appetite stimulation). Yet, the effects and mechanisms of cannabis on HIV associated chronic inflammation, the endocannabinoid system, immune modulation and neurologic disorders are minimally understood. As indicated in the RFA, preclinical models can provide a rigorous in-depth analysis of the molecular and cellular mechanisms at the intersection between phytocannabinoids, HIV and ART. To this end, we propose a comprehensive evaluation of the two most used cannabinoid compounds (THC, CBD) on BBB function, immune-endothelial interactions and neuroinflammation. We will utilize state of the art chip microfluidics models of the neurovascular unit (NVU) and animal models for HIV (w/ w/o ART). Previously, we discovered that the brain endothelium upregulate CB2 in HIV infected human brain tissue. We have also found that modulation of CB2 affects indices of HIV pathology (in-vivo) and regulates the BBB. Our preliminary studies identify the diverse effects that phytocannabinoids can have on the different properties of the BBB. Specifically, cannabinoids (THC, CBD) alone can enhance the physical barrier, partially reduce endothelial activation and augment efflux transporter activity. Although some of these effects may appear beneficial, the presence of HIV and ART changes how the function of the BBB is regulated by cannabinoid substances. For example, the augmented transporter activity by THC has important considerations for altering ART-CNS penetrability. Thus, we hypothesize that phytocannabinoids differentially modulates BBB function that are both beneficial and deleterious in NeuroHIV. In Aim 1, using our latest tissue-engineered microfluidic NVU model, we will perform analyses of the kinetic changes in BBB permeability, transporter status and immune-endothelial interaction. Then, in Aim 2, we will compare outcomes between widely used routes of cannabinoid administration (oral vs. inhaled) in vivo using two relevant models of HIV infection (‘humanized’ mice and a model of aseptic meningitis/encephalitis). Experiments will evaluate changes in the BBB in the context of ART and cannabinoid exposure. Finally, we propose to identify novel crosstalk mechanisms that bridge cannabinoid receptor signaling to signals that control BBB maintenance (Aim 3). It’s clear that cannabinoids exert unknown cell specific effects that contribute to the tumultuous interpretation of how these compounds impact NeuroHIV. Using innovative preclinical tools, our studies will contribute significantly towards understanding the consequences of cannabinoid use on the BBB in the modern era of NeuroHIV.