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

描述（由申请人提供）：已知 HIV 感染者长期饮酒的增加会增加 HIV-1 复制并降低对高效抗逆转录病毒疗法 (HAART) 的反应。众所周知，酒精会破坏血脑屏障，进一步增加感染艾滋病毒的单核细胞/巨噬细胞（主要病毒库）对中枢神经系统的浸润，从而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 HAART 药物反应降低（尤其是在单核细胞/巨噬细胞中）的潜在机制尚不清楚。据报道，通过各种兴奋剂/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 可以代谢重要的 HAART 药物、蛋白酶抑制剂 (PI)，因此 CYP3A4 的作用被认为在确定 PI 的生物利用度和功效方面至关重要。此外，已知 CYP3A4 会被乙醇和 PI 诱导，这可能会导致 PI 代谢进一步增加，从而降低其功效。关于 CYP 途径在酒精介导的氧化损伤、HIV-1 复制和单核细胞/巨噬细胞中 PI 代谢改变中的直接作用知之甚少。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 复制和单核细胞/巨噬细胞对 HAART 反应中的作用，及其在神经艾滋病发病机制中的影响。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 PI 功效降低和 PI 介导的毒性增加以及 HIV-1 复制速率的增加是通过 CYP3A4 直接介导的。为了检验我们的假设，我们提出两个目标。目标 1：检查 CYP2E1 和 CYP3A4 在单核细胞/巨噬细胞中乙醇介导的 PI 和 HIV-1 复制功效中的贡献。目标 2：确定来自酒精性 HIV 感染者的单核细胞/巨噬细胞特有的细胞、生化和分子变化。成功完成拟议的研究后，我们期望确定 CYP2E1 参与乙醇介导的氧化应激和 HIV-1 复制，并且酒精暴露会改变 PI-CYP3A4 相互作用，从而降低酒精 HIV 个体中 PI 的功效。这些发现将为了解酗酒者/HIV 个体中 HIV-1 发病机制和 HAART 治疗策略开辟一条新途径。

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.