Epigenetic mechanisms underlying cannabinoid modulation of neuroinflammation in HIV/SIV infection

摘要 HIV相关性神经疾病(HAND)，一种主要的共病，影响约30%-50%的接受治疗的患者 抑制性抗逆转录病毒治疗的特点是注意力、注意力集中、决策困难。 以及记忆力、抑郁和动作减慢。尽管脑小胶质细胞的慢性激活被认为是 驾驶之手，分子机制仍然不清楚。新出现的证据表明，表观遗传学 涉及DNA异常甲基化的机制可能在多发性硬化的发病机制中起重要作用 硬化症、帕金森氏症和阿尔茨海默氏症。然而，DNA甲基化在HIV中的作用 诱导的单核细胞/小胶质细胞激活仍不清楚。表观遗传标记是可遗传的，而且 在有丝分裂过程中传递给几代子细胞可能解释了一种潜在的机制 单核细胞/小胶质细胞的激活反过来可能有助于维持CART治疗后的持续性神经炎症 病人。我们的初步研究发现，促炎症干扰素刺激和 基底节趋化因子基因和启动子CpG岛DNA甲基化显著改变 与炎症反应、细胞凋亡、dsDNA损伤反应和氧化应激相关的基因 慢性SIV感染猕猴的结肠上皮细胞。更重要的是，慢性大麻素 对感染ART SIV的恒河猴的治疗可阻止大脑和大脑中促炎基因的表达 表观遗传学改变表明它们在减轻神经炎症和 减少与手有关的症状。在拟议的研究中，我们将首次调查DNA的变化 β-9-四氢大麻酚(THC)、大麻二酚、CB2R激动剂JWH133诱导的甲基化 在SIV感染过程中抑制单核细胞/小胶质细胞的激活。此外，我们将确定 联合抗逆转录病毒治疗(CART)联合慢性大麻素治疗的疗效观察 表观遗传改变，病毒库，炎症和内源性大麻素水平在血液和 脑脊液。最后，我们将研究受体介导的THC的分子机制。 阻断内质网应激，这是神经退行性疾病发病的关键事件。建议数 研究具有高度创新性，并应用最先进的免疫学和分子方法来填补 在我们对与手相关的表观遗传机制的理解上存在显著差距。就像大麻素一样 在神经疾病的治疗方面显示出巨大的希望，拟议的研究是必要的，它将 对其抗肿瘤作用的表观遗传学和内源性大麻素机制有一个基本的了解 发炎效应。最后，这些结果将对免疫调节具有重要的治疗意义。 不仅是艾滋病毒，还有其他慢性神经炎症性疾病。

ABSTRACT HIV associated neurological disorder (HAND), a major comorbidity affecting about 30-50% of patients receiving suppressive anti-retroviral therapy is characterized by difficulties with attention, concentration, decision making and memory, depression and slowed movements. Although chronic activation of brain microglia is proposed to drive HAND, the molecular mechanisms remain ill defined. Emerging evidence has shown that epigenetic mechanisms involving aberrant DNA methylation may significantly contribute to the pathogenesis of multiple sclerosis, Parkinson’s and Alzheimer’s disease. Nevertheless, the role of DNA methylation specifically in HIV induced monocyte/microglia activation remains unknown. The fact that epigenetic marks are heritable and passed on to several generations of daughter cells during mitosis might explain a potential mechanism causing monocyte/microglial activation that may in turn help maintain persistent neuroinflammation in cART treated patients. Our preliminary studies identified significant upregulation of proinflammatory interferon stimulated and chemokine genes in basal ganglia and marked alterations in DNA methylation of CpG islands in promoters of genes associated with inflammatory response, apoptosis, dsDNA damage response and oxidative stress in colonic epithelium of chronically SIV-infected macaques, respectively. More importantly, chronic cannabinoid treatment to ART naïve SIV-infected rhesus macaques prevented proinflammatory gene expression in brain and epigenetic alterations suggesting their immense therapeutic potential for attenuating neuroinflammation and reduced HAND related symptoms. In the proposed studies, we will for the first time investigate changes in DNA methylation associated with delta-9-tetrahydrocannabinol (THC), cannabidiol, JWH133 (CB2R agonist) induced suppression of monocyte/microglial activation through the course of SIV infection. Further, we will determine the effect of combination anti-retroviral treatment (cART) in conjunction with chronic cannabinoid treatments on epigenetic alterations, viral reservoir, inflammation and endogenous cannabinoid levels in the blood and cerebrospinal fluid. Finally, we will investigate the receptor mediated and molecular mechanisms by which THC blocks endoplasmic reticulum stress, a key event in the onset of neurodegenerative diseases. The proposed research is highly innovative and applies state of the art immunological and molecular approaches to fill a significant gap in our understanding of the epigenetic mechanisms associated with HAND. As cannabinoids have shown great promise for the treatment of neurological disorders, the proposed studies are necessary, as it will provide a fundamental understanding of the epigenetic and endocannabinoid mechanisms underlying their anti- inflammatory effects. Finally, the results will have important therapeutic implications for immune modulation in not only HIV but also other chronic neuroinflammatory diseases.