Glial-neuronal interactions underlying the molecular feedback between HIV viral p

描述（由申请人提供）：本研究的长期目标是描述人类免疫缺陷病毒1 （HIV）阳性个体对甲基苯丙胺（METH）等物质滥用增加的可能机制。在这个为期两年的探索性项目中，我们建议专门研究甲基苯丙胺（甲基安非他明），一种成瘾的精神兴奋剂，和HIV-1病毒蛋白对大鼠脑胶质神经元炎症诱导的协同作用。在中枢神经系统（CNS），神经胶质细胞被HIV-1病毒感染，引起神经炎症。脑部炎症增加与神经退行性疾病和行为障碍相关，如HIV-1诱导的神经退行性疾病（HAND）和HIV-1诱导的痴呆（HAD）。此外，艾滋病毒感染者滥用冰毒可导致病毒载量增加和严重的脑相关疾病。在这些研究中，我们将使用非感染性HIV-1转基因（HIV-1Tg）大鼠和原代细胞培养模型。HIV-1Tg大鼠携带gag/pol缺失的原病毒基因，表达9种HIV-1病毒蛋白中的7种。这只HIV- 1tg大鼠表现出人类HIV疾病的临床表现，因此，模仿了在接受抗逆转录病毒治疗的HIV患者体内持续存在HIV-1蛋白所导致的感染。最近，通过PCR阵列分析，我们发现与F344对照大鼠相比，HIV-1Tg大鼠大脑中的促炎细胞因子IL-1¿以及Ccl2， Ccl3和Ccl7趋化因子增加了更大程度。我们还证明了HIV-1Tg大鼠前额皮质多巴胺D1受体（D1R）表达升高，并且这些动物有更大的甲基安非他明诱导的行为敏感，通过刻板的头部运动来评估。基于文献和我们之前的数据，我们假设在HIV-1病毒蛋白存在的情况下，滥用物质，如冰毒，会增强神经胶质的激活，从而影响神经元，增加多巴胺能系统的活性，从而增加冰毒的摄入量。为了验证这一假设，我们将首先使用免疫组织化学方法检测暴露于甲基安非他明和未暴露于甲基安非他明的HIV-1Tg大鼠大脑中神经胶质细胞与神经元细胞的相互作用。然后，我们将确定神经递质活性的基因表达变化，特别是多巴胺能系统，以及神经炎症相关标志物。我们还将从HIV-1Tg大鼠中分离出原代胶质细胞和神经元细胞，并检测多巴胺能和其他神经递质途径以及炎症、凋亡和氧化应激标志物在体外暴露于甲基安非他明后的基因表达变化。我们的具体目标包括：1)确定甲基安非他明和HIV病毒蛋白对HIV-1Tg大鼠脑内神经胶质和神经元激活、神经炎症和神经递质调节的影响；2)利用从HIV-1Tg大鼠脑内分离的原代神经胶质和神经元细胞，在离体模型中确定甲基安非他明和HIV病毒蛋白对神经胶质和神经元激活、神经炎症和神经递质调节的影响。这个提议代表了第一个探索甲基安非他明对神经胶质相互作用影响的项目

DESCRIPTION (provided by applicant): The long-term goal of this research is to delineate the possible mechanisms underlying the increased abuse of substances, such as methamphetamine (METH), by individuals who are human immunodeficiency virus-1 (HIV)-positive. In this two-year exploratory project, we propose to specifically examine the synergistic effects of methamphetamine (METH), an addictive psychostimulant, and HIV-1 viral proteins on glial-neuronal induction of inflammation in the rat brain. In the central nervous system (CNS), the glial cells are infected by the HIV-1 virus, causing neuroinflammation. Increased inflammation in the brain has been correlated with neurodegenerative diseases and behavioral disorders, such as HIV-1 induced neurodegeneration (HAND) and HIV-1 induced dementia (HAD). Moreover, METH abuse in the HIV-infected individual can lead to increased viral loads and severe brain-related disorders. For these studies, we will use both the non-infectious HIV-1 transgenic (HIV-1Tg) rat and primary cell culture models. The HIV-1Tg rat carries a gag/pol deleted provirus gene, and expresses 7 out of 9 HIV-1 viral proteins. This HIV-1Tg rat develops clinical manifestations of human HIV disease, and, thus, mimics the infection that results from the persistent presence of HIV-1 proteins in HIV patients given anti-retroviral therapy. Recently, using PCR array analysis, we showed that the pro- inflammatory cytokine, IL-1¿, as well as the Ccl2, Ccl3, and Ccl7 chemokines, are increased to a greater extent in the brain of the HIV-1Tg rat compared to the F344 control rat. We have also demonstrated that there is elevated dopamine D1 receptor (D1R) expression in the prefrontal cortex of HIV-1Tg rats, and that these animals have greater METH-induced behavioral sensitization, as assessed by stereotypical head movement. Based on the literature and our previous data, we hypothesize that the abuse of substances, such as METH, in the presence of HIV-1 viral proteins, enhances glial activation, which affects neurons, and increases the activity of the dopaminergic system, thereby increasing the intake of METH. To test this hypothesis, we will first examine the glial-neuronal cell interaction in the brain of the HIV-1Tg rat with and without exposure to METH using immunohistochemistry. We will then identify gene expression changes in neurotransmitter activity, in particular, the dopaminergic system, as well as neuroinflammation-related markers. We will also isolate primary glial and neuronal cells from HIV-1Tg rats and examine gene expression changes in the dopaminergic and other neurotransmitter pathways, as well as inflammation, and apoptotic and oxidative stress markers after in vitro exposure to METH. Our specific aims include: 1) To determine the effects of METH and HIV viral proteins on glial and neuronal activation, neuroinflammation, and neurotransmitter modulation in the brain of the HIV-1Tg rat, and 2) To determine the effects of METH and HIV viral proteins on glial and neuronal activation, neuroinflammation, and neurotransmitter modulation in an ex vivo model using primary glial and neuronal cells isolated from the HIV- 1Tg rat brain To our knowledge, this proposal represents the first project to explore the effects of METH on glial-neuronal interactions and the subsequent effects on the dopaminergic system using the HIV-1Tg rat and primary cell cultures isolated from the HIV-1Tg rat. The data from this research will shed light on possible cellular and molecular mechanisms underlying the increased abuse of substances such as METH by HIV-positive individuals, and help to elucidate the glial-neuronal interactions associated with METH abuse and neuroAIDS. Our exploratory studies can also lead to better treatment approaches that will ultimately benefit both HIV-1 patients and substances abusers. Thus, the proposed studies have high clinical relevance and will contribute significantly to the understanding and treatment of neurological complications associated with substance abuse, HIV infection, and AIDS.