Role of cytochrome P450 in alcohol-mediated effects on antiretroviral and HIV-1

描述（由申请人提供）：已知hiv感染者慢性饮酒的增加会增加HIV-1复制并降低对高效抗逆转录病毒治疗（HAART）的反应。已知酒精还会破坏血脑屏障，进一步增加受hiv感染的单核/巨噬细胞（主要病毒库）在中枢神经系统的浸润，导致神经艾滋病的发展。然而，酒精暴露导致HIV-1复制增加和对HAART药物反应降低的潜在机制，特别是在单核/巨噬细胞中，尚不清楚。据报道，通过各种刺激物/途径产生的氧化应激是导致HIV-1复制增加的原因。肝脏中主要的慢性酒精介导的氧化应激途径是乙醇诱导的细胞色素P450 2E1 （CYP2E1）的表达。由于已知CYP3A4代谢重要的HAART药物，蛋白酶抑制剂（PIs）， CYP3A4的作用被认为是决定PIs的生物利用度和疗效的关键。此外，已知CYP3A4可被乙醇和PI诱导，这可能导致PI的代谢进一步增加，从而降低其功效。对于CYP通路在酒精介导的氧化损伤、HIV-1复制和单核/巨噬细胞中pi代谢改变中的直接作用，人们知之甚少。我们的长期目标是确定CYP通路在酒精介导的氧化应激、HIV-1复制和单核/巨噬细胞对HAART的反应中的作用，以及它们在神经艾滋病发病机制中的意义。我们的中心假设是，在单核细胞/巨噬细胞中，酒精介导的氧化应激通过CYP2E1介导的途径导致HIV-1复制增强。此外，酒精介导的PIs疗效下降和PIs介导的毒性增加同时HIV-1复制率增加是通过CYP3A4直接介导的。为了验证我们的假设，我们提出了两个目标。目的1：检测CYP2E1和CYP3A4在乙醇介导的PI疗效和单核/巨噬细胞中HIV-1复制中的作用。目的2：确定酒精性hiv感染患者单核/巨噬细胞的细胞、生化和分子变化。在成功完成所提出的研究后，我们期望已经确定CYP2E1参与乙醇介导的氧化应激和HIV-1复制，并且酒精暴露改变PI- cyp3a4相互作用，从而降低PI在酒精性HIV+个体中的疗效。这些发现将为理解HIV-1发病机制和酒精/HIV+个体的HAART治疗策略开辟新的途径。

DESCRIPTION (provided by applicant): An increase in chronic alcohol consumption in HIV-infected individuals is known to increase HIV-1 replication and decrease responses to highly active antiretroviral therapy (HAART). Alcohol is also known to disrupt blood-brain barrier, which further increases the CNS infiltration of HIV-infected monocytes/ macrophages (major viral reservoir), leading to development of neuroAIDS. However, the underlying mechanism(s) by which alcohol exposure results in increased HIV-1 replication and decreased response to HAART drugs, especially in monocytes/macrophages, is not known. Oxidative stress generated through various stimulants/pathways has been reported to be responsible for increased HIV-1 replication. The major chronic alcohol-mediated oxidative stress pathway in the liver is ethanol-induced expression of cytochrome P450 2E1 (CYP2E1). Since CYP3A4 is known to metabolize important HAART drugs, protease inhibitors (PIs), the role of CYP3A4 is considered critical in determining the bioavailability and efficacy of PIs. In addition, CYP3A4 is known to be induced by ethanol and PI, which could result in further increases in metabolism of PIs, thereby, decreasing their efficacy. There is relatively little known about the direct contribution of CYP pathways in alcohol-mediated oxidative damage, HIV-1 replication, and altered metabolism of PIs in monocytes/macrophages. Our long-term goal is to define the role of CYP pathways in alcohol-mediated oxidative stress, HIV-1 replication, and response to HAART in monocytes/macrophages, and their implications in pathogenesis of neuroAIDS. Our central hypothesis is that in monocytes/macrophages alcohol-mediated oxidative stress causes enhanced HIV-1 replication via pathways mediated through CYP2E1. In addition, alcohol-mediated decrease in the efficacy of PIs and increase in PIs-mediated toxicity concurrent with increase in the rates of HIV-1 replication is directly mediated through CYP3A4. To test our hypothesis, we propose two aims. Aim 1: Examine the contribution of CYP2E1 and CYP3A4 in ethanol-mediated efficacy of PI and HIV-1 replication in monocytes/ macrophages. Aim 2: Determine cellular, biochemical, and molecular changes unique to monocytes/ macrophages derived from alcoholic HIV-infected patients. Upon successful completion of the proposed research, we expect to have established that CYP2E1 is involved in ethanol-mediated oxidative stress and HIV-1 replication and that alcohol exposure alters PI-CYP3A4 interaction, thereby, decreasing the efficacy PI in alcoholic HIV+ individuals. These finding will open a new avenue in understanding HIV-1 pathogenesis and HAART treatment strategy among alcoholic/HIV+ individuals.