Inter- and Intra-cellular effects of cannabinoids, HIV and ART in the CNS

摘要： 抗逆转录病毒疗法(ART)极大地延长了艾滋病毒携带者(PLWH)的生命；然而， 他们继续经历过多的共病，包括神经元紊乱和疼痛。 40%到72%的PLWH使用大麻来缓解焦虑、压力、ART副作用、疼痛和/或 超过55%的患者至少每天使用大麻带来的愉悦。有趣的是，最近的一项研究发现，人们 大量使用大麻的人在抗逆转录病毒治疗中减少了PLWH中的炎症特征。这些和无数的 其他研究支持植物大麻素在一些疾病中的抗炎和免疫调节作用 器官系统，包括心脏、结肠、肾脏、肝脏和肠道；然而，它们的药用价值受到 精神药物活动。将抗炎作用与精神作用分开的努力已经 揭示了包括大麻受体1(CB2)CB2在内的3种内源性受体的不同活性 表现出与内源性和植物性大麻素不同的组织表达和兴奋性。有几份报告说 研究表明，大麻素可以减少艾滋病毒在T细胞、巨噬细胞、树突状细胞和 人胎小胶质细胞体外培养。然而，大麻素对小胶质细胞感染HIV的影响 ART的背景和中枢神经系统中邻近神经元和星形胶质细胞的正常细胞环境 被检查过了。几项研究明确表明CB2激动剂具有抗高血压作用。 心脏、肠道、实验性自身免疫性脑炎和神经病理性疼痛的炎症特性 炎性小体激活。这导致我们假设大麻素信号影响艾滋病毒感染。 以及中枢神经系统存在ARV时的慢性炎症 炎症者。 为了在中枢神经系统细胞的背景下检查艾滋病毒感染，我们开发了一种人类 诱导多能干细胞由神经元、星形胶质细胞和小胶质细胞组成的三种培养模型。这款车 概括了中枢神经系统艾滋病毒感染的几个关键方面，包括细胞因子的产生增加， 氧化应激反应、炎症信号和综合应激反应。抗逆转录病毒治疗减少 HIV感染和炎症信号通路；然而，尽管 病毒抑制。我们建议进一步发展这一模型，以确定大麻类物质 通过减少炎症小体来调节HIV诱导的炎症和随后的神经元功能障碍 通过以下方式激活：1)确定大麻素对慢性艾滋病毒感染和抗逆转录病毒疗法的影响 IMgl/iNrn/iAstr三元培养。2)测定大麻素对细胞因子水平、炎症因子的影响 在iMg1/iNrn/iAstr三元培养中，表达谱和小胶质细胞激活。3)确定 大麻素在HIV感染中对神经元和星形胶质细胞的影响以及在iMgl/iNrn/iAstr培养中的作用。

Abstract: Antiretroviral therapy (ART) has dramatically extended the lives of people living with HIV (PLWH); however, they continue to experience a plethora of co-morbid conditions including neuronal disorders and pain. Between 40 and 72% of PLWH use cannabis to mitigate anxiety, stress, ART side effects, pain, and/or for pleasure with over 55% of patients using cannabis at least daily. Interestingly, a recent study found that people who use cannabis heavily had reduced inflammatory signatures in PLWH on ART. These and a numerous other studies support the anti-inflammatory and immunomodulator effects of phytocannabinoids in a number of organ systems including heart, colon, kidney, liver and the gut; however, their medicinal use is confounded by the psychotropic activities. Efforts to separate the anti-inflammatory effects from the psychotropic effects have revealed differential activities of 3 endogenous receptors including cannabis receptor 1 (CB2) CB2 which exhibit differential tissue expression and agonism with endo- and phyto-cannabinoids. Several reports have shown that cannabinoids attenuate HIV infection and/or replication in T-cells, macrophages, dendritic cells and human fetal microglia cultured ex vivo. However, the effect of cannabinoids on HIV infection of microglia in the context of ART and the normal cellular environment of neighboring neurons and astrocytes in the CNS has not been examined. Several studies specifically implicate CB2 agonism which has been shown to have anti- inflammatory properties in the heart, gut, experimental autoimmune encephalitis and neuropathic pain via inflammasome activation. This has led us to hypothesize that Cannabinoid signaling influences HIV infection and chronic inflammation in the presence of ARV in the central nervous system by attenuating the inflammasome. In order to examine HIV infection in the context of cells of the CNS, we have developed a human induced pluripotent stem cell tri-culture model composed of iNeurons, iAstrocytes, and iMicroglia. This model recapitulates several key aspects of HIV infection in the CNS including increased cytokine production, oxidative stress response, inflammatory signaling, and integrated stress response. ARV treatment reduces HIV infection and inflammatory signaling pathways; however, a subset of pathways remain elevated despite viral suppression. We propose to further develop this model to determine the ability of cannabinoids to modulate HIV-induced inflammation and subsequent neuronal dysfunction via reducing inflammasome activation by: 1) Determining the effect of cannabinoids on chronic HIV infection and ART in the context of iMgl/iNrn/iAstr triculture. 2) Determining the effect of cannabinoids on cytokine levels, inflammatory gene expression profile, and microglial activation in iMgl/iNrn/iAstr triculture. 3) Determining the effect of cannabinoids on neurons and astrocytes in HIV infection and ART in iMgl/iNrn/iAstr triculture.