Attenuation of astroglial chemokine expression by beta-funaltrexamine: implicatio

描述(申请人提供)：全球有4000多万人感染了人类免疫缺陷病毒(HIV-1)。大约10-15%的HIV-1感染者患有中枢神经系统疾病，包括HIV相关性脑炎和HIV痴呆(HAD)，统称为神经艾滋病。越来越多的证据表明，与神经艾滋病相关的神经元损伤部分是由小胶质细胞和星形胶质细胞介导的炎症引起的。星形胶质细胞衍生的趋化因子，包括CCL2和CXCL10，被认为是参与神经艾滋病的炎症分子。我们最近研究了2-FNA抑制星形胶质细胞趋化因子表达的作用，然而，2-FNA抑制趋化因子表达的机制尚不清楚。因此，迫切需要确定2-FNA抑制HIV-1诱导的星形胶质细胞趋化因子表达的机制。长期目标是确定2-FNA抗炎作用的分子机制。该应用的总体目标是确定2-FNA减弱星形胶质细胞CCL2和CXCL10表达的机制。中心假说是2-FNA部分通过转录前机制抑制星形胶质细胞CCL2和CXCL10的表达。这一假设将通过包括三个不同目标的实验来验证：特定目标1：确定2-Funaltrexamine对促炎诱导的人星形胶质细胞CCL2和CXCL10表达的影响。特异性目的2：确定阿片受体在2-FU介导的促炎症诱导人星形胶质细胞CCL2和CXCL10表达中的作用。特异性目的3：研究2-Funaltrexamine对人脑星形胶质细胞NFkB活化的影响。为了模拟与HIV-1感染相关的中枢神经系统炎症的一个关键方面，我们将人星形胶质细胞暴露于TNF1和HIV-1Tat1-72，从而诱导CCL2和CXCL10。然后，我们将研究2-FNA对星形胶质细胞CCL2和CXCL10表达的影响。此外，还将使用选择性阿片受体拮抗剂，以确定特定阿片受体类型参与2-FNA的抗炎作用。氧化应激和转录因子NFkB在促炎诱导的CCL2和CXCL10的表达中起重要作用。因此，2-FNA对活性氧物种的产生和NFkB活化的影响也将被表征。该项目将通过识别和表征阿片类拮抗剂2-FNA的新作用，特别是该分子的抗炎作用，来填补我们知识库中的基本空白。这些信息有望有助于制定治疗策略，以预防和治疗神经炎性疾病，包括艾滋病毒-1神经发病机制。与公共卫生的相关性通过这一领域奖获得的信息预计将有助于制定治疗战略，以预防和治疗多种神经炎性疾病，特别是艾滋病毒-1神经发病机制。此外，该奖项将支持俄亥俄州立大学CHS的多名本科生、研究生和医学生的研究培训。这项培训将有助于这些学生在未来的健康相关职业生涯中取得成功。

DESCRIPTION (provided by applicant): Worldwide, there are more than 40 million people infected with human immunodeficiency virus (HIV-1). Approximately 10-15% of HIV-1 infected individuals suffer from CNS pathologies including HIV-associated encephalitis and HIV dementia (HAD), collectively termed neuroAIDS. Increasing evidence suggests that neuronal damage associated with neuroAIDS results in part from microglial and astroglial mediated inflammation. Astrocyte-derived chemokines, including CCL2 and CXCL10, have been implicated as inflammatory molecules involved in neuroAIDS. We recently characterized 2-funaltrexamine (2-FNA) inhibition of astroglial chemokine expression; however, the mechanism by which 2 FNA inhibits chemokine expression remains unclear. A critical need exists, therefore, to identify the mechanism whereby 2-FNA inhibits HIV-1-induced astroglial chemokine expression. The long-term goal is to identify the molecular mechanism responsible for the anti-inflammatory actions of 2-FNA. The overall objective of this application is to identify the mechanism by which 2-FNA attenuates astroglial expression of CCL2 and CXCL10. The central hypothesis is that 2-FNA inhibits astroglial expression of CCL2 and CXCL10 in part through a pre-transcriptional mechanism. This hypothesis will be tested through experiments included in three separate aims: SPECIFIC AIM 1: Determine the effects of 2-funaltrexamine on proinflammatory-induced CCL2 and CXCL10 expression in human astroglia. SPECIFIC AIM 2: Determine the role of opioid receptors in 2-funaltrexamine mediated effects on proinflammatory induced CCL2 and CXCL10 expression in human astroglia. SPECIFIC AIM 3: Determine the effects of 2-funaltrexamine on NFkB activation in human astroglia. To model a key aspect of CNS inflammation associated with HIV-1 infection, we will expose human astroglial cells to TNF1 and HIV-1 Tat1-72 resulting in the induction CCL2 and CXCL10. We will then characterize the effects of 2-FNA on CCL2 and CXCL10 expression in astroglia. Also, selective opioid receptor antagonists will be utilized in order to characterize the involvement of specific opioid receptor types in the anti inflammatory actions of 2-FNA. Oxidative stress and the transcription factor NFkB are instrumental in the proinflammatory induced expression of both CCL2 and CXCL10. Thus, the effects of 2-FNA on the generation of reactive oxygen species and NFkB activation will also be characterized. This project will fill basic gaps in our knowledge base by identifying and characterizing novel actions of the opioid antagonist 2-FNA, particularly the anti-inflammatory actions of this molecule. This information is expected to be instrumental in the development of therapeutic strategies to prevent and treat neuroinflammatory conditions including, HIV-1 neuropathogenesis. RELEVANCE TO PUBLIC HEALTH The information obtained through this AREA Award is expected to be instrumental in the development of therapeutic strategies to prevent and treat numerous neuroinflammatory conditions and in particular, HIV-1 neuropathogenesis. Additionally, this award will support the research training of multiple undergraduate, graduate and medical students at OSU-CHS. This training will lend to the success of these students in their future careers in health related professions.