Drugs of Abuse and NeuroAIDS: Proteome Activity Profiles

描述(申请人提供)：在引入高度有效的抗逆转录病毒疗法(HAART)以来的十年里，大多数发达社会的大多数患者的艾滋病临床病程的严重性都得到了改善。然而，在发展中世界，艾滋病的发病率和临床严重性继续呈指数增长，部分原因是静脉注射毒品导致无意中接触到艾滋病毒-1病毒。在斯克里普斯研究所现有的NIDA支持的项目(DA12444)中，我领导了一系列研究，使用了几种神经艾滋病的动物模型，调查病毒诱导的神经致病作用和滥用药物之间的潜在毒性相互作用。我们现在有强有力的证据表明，在这些动物模型中，甲基苯丙胺诱导的毒性和神经艾滋病的进展之间存在积极的相互作用，即协同作用。这种协同作用背后的机制尚不明显，但部分可能涉及直接有毒的病毒产物(即gp120)与针对神经元和/或胶质的宿主衍生因子的相互作用，以及与药物相关的氧化应激和氧化还原平衡丧失所导致的受损细胞过程的相互作用。 在这项CEBRA申请中，我们建议启动一系列新的研究，从概念上扩展目前的研究路线，更好地阐明导致病毒产品、宿主衍生因子和滥用药物之间的毒性协同作用的细胞和分子机制。拟议的研究计划涉及一种非常新的化学方法，该方法利用一种新的蛋白质组学分析形式，其中特定蛋白质(酶)的激活状态将在两种公认的在甲基苯丙胺暴露的情况下与艾滋病相关的神经发病的动物模型中表征和量化。这些新方法是由斯克里普斯研究所首创的，它们利用了这样一个事实，即特定酶家族的激活状态比蛋白质本身的水平更准确地反映了蛋白质的功能状态。在星形细胞中过度表达HIV-1病毒外壳蛋白gp120或宿主衍生的免疫相关细胞因子IL-6的基因工程小鼠，将被用来比较和对比这两个独立的艾滋病相关神经发病模型中涉及的酶的活性状态。这些研究将包括对这些神经艾滋病模型的第一次蛋白质组活性分析，并将提供甲基苯丙胺引起的神经毒性协同作用的独特分子图谱。

DESCRIPTION (provided by applicant): In the decade since the introduction of highly affective anti-retroviral therapy (HAART), the severity of the clinical course of AIDS has been ameliorated for a majority of afflicted individuals in most developed societies. However, in the developing world the incidence and clinical severity of AIDS continues in an exponential fashion fueled, in part, by inadvertent HIV-1 exposure through intravenous drug use. In an existing NIDA-supported Program Project at the Scripps Research Institute (DA12444), I direct a series of studies employing several animal models of neuroAIDS that investigate the potential toxic interaction between viral-induced neuropathogenesis and drugs of abuse. We now have strong evidence of a positive interaction, i.e. synergy, between methamphetamine-induced toxicity and the progression of NeuroAIDS in these animal models. The mechanisms underlying this synergy are not apparent but may, in part, involve interactions of directly toxic viral products (i.e. gp120) with host-derived factors targeting neurons and/or glib, and with compromised cellular processes precipitated by drug-related oxidative stress and loss of redox poise. In this CEBRA application we propose to initiate a new series of studies that will conceptually extend the current line of investigation that could better elucidate the cellular and molecular mechanisms that lead to the toxic synergism between viral products, host-derived factors and drugs of abuse. The proposed research plan involves a very new chemical approach that utilizes a novel form of proteomic analysis in which the activation state of specific proteins (enzymes) will be characterized and quantified in two accepted animal models of AIDS-related neuropathogenesis in the context of methamphetamine exposure. These new methods, pioneered at The Scripps Research Institute, take advantage of the fact that activation states of specific enzyme families are a more accurate reflection of the functional state of proteins than are protein levels per se. Genetically engineered mice over-expressing either the HIV-1 viral coat protein, gp120, or the host-derived, immune-related cytokine, IL-6, in astrocytes, will be used to compare and contrast the activity states of enzymes involved in these two independent models of AIDS-related neuropathogenesis. These studies will comprise the first proteome activity analysis of these of neuroAIDS models, and will provide a unique molecular profile of the neurotoxic synergy elicited by methamphetamine.