HIV-Induced Dopamnergic Changes and Methamphetamine Toxicity Mediated through Mic

描述(由申请人提供)：这是一个响应PAR-10-021在艾滋病毒/艾滋病药物滥用研究的广泛领域的试点项目。该项目的目标是了解艾滋病毒感染的巨噬细胞如何与甲基苯丙胺(Meth)结合导致多巴胺能神经细胞的变化，并确定靶向microRN(MiR)是否可以逆转或预防功能障碍。滥用药物，如甲基苯丙胺和可卡因，在化学上类似于多巴胺，通过逆转多巴胺转运体而导致大脑中的多巴胺能功能障碍，从而激活多巴胺受体，并导致突触空间中高浓度的多巴胺积累。此外，甲基苯丙胺导致多巴胺能神经元囊泡性单胺转运体功能障碍，导致囊泡性多巴胺包装功能障碍，胞浆中多巴胺积聚，下游氧化应激。人们还知道，艾滋病毒和病毒在大脑中的感染会导致长期感染的大脑中的多巴胺能细胞丢失，尽管其机制尚不清楚。艾滋病毒感染者中的药物滥用和吸毒成瘾对公共健康构成重大威胁，并使人们对药物滥用背后的基本生化机制以及艾滋病毒/艾滋病的病因和发病机制有了新的认识。根据研究人员产生的数据，我们假设Meth和HIV结合在一起可以诱导DA神经元的miR变化，特别是miR-9、miR-124、miR-125a、miR-130b和let-7d；同时对DA合成、代谢和运输造成不利，这些影响可以通过增加特定的miR来预防或逆转。围绕这一假说，我们的目标是：1.了解Meth和HIV对miR表达、DA合成、代谢和转运的联合影响；2.通过外源性改变miR功能来干预预防HIV和Meth引起的多巴胺能缺陷。我们的研究模型将是细胞培养：SH-SY5Y细胞和感染艾滋病毒的单核细胞。我们的方法是使用功能损失和功能增益技术。通过过度表达或抑制特定的miRs以及检测多巴胺代谢物、转运体和酶，我们可以了解miR如何介导HIV引起的多巴胺能功能障碍。该项目旨在通过更好地了解与滥用药物和艾滋病毒/艾滋病疾病过程的相互作用，改善吸毒者的艾滋病毒治疗和结果。该项目的结果将进一步加深我们对影响药物滥用风险和保护因素的基因(通过miR的调节)和环境(HIV感染)的理解。知识将被应用于帮助设计逆转或预防艾滋病毒/艾滋病化学成瘾的疗法。这一项目的结果将用于今后对药物滥用和艾滋病毒/艾滋病药物以及用于治疗艾滋病毒的药物中微效受体的作用的研究。未来的研究还包括研究MIR在艾滋病毒/艾滋病中的作用以及药物滥用和成瘾的潜在神经生物学。 公共卫生相关性：甲基苯丙胺滥用和人类免疫缺陷病毒(HIV)感染是重大的公共卫生风险，两者结合在一起产生了“双重流行”。这个项目试图了解当神经元暴露在这两种侮辱下时，发生的分子遗传交集和神经化学变化。这一研究结果将有助于更深入地了解HIV和甲基苯丙胺滥用中神经化学变化的分子机制，并为治疗药物滥用人群中HIV的病原学和神经发病机制提供可能的治疗干预措施。

DESCRIPTION (provided by applicant): This is a pilot project in response to PAR-10-021 in the broad area of drug abuse research in HIV/AIDS. The goal of the project is to understand how HIV-infected macrophages combine with methamphetamine (Meth) to cause alterations in dopaminergic neural cells and determine whether targeting a microRN (miR) can reverse or prevent dysfunction. Drugs of abuse, such as methamphetamine and cocaine, are chemically similar to dopamine and cause dopaminergic dysfunction in the brain by reversing the dopamine transporter and thus activating dopamine receptors and cause accumulation of high concentrations of dopamine in the synaptic space. Further, methamphetamine causes malfunction of the vesicular monamine transporter in dopaminergic neurons, leading to dysfunctional packaging of vesicular dopamine, accumulation of dopamine in the cytoplasm and downstream oxidative stress. It is also known that HIV and viral infection in the brain lead to dopaminergic cell loss in the long-term infected brain, though the mechanism is not well understood. Drug abuse and drug addiction in the HIV-infected population pose significant public health risks and new knowledge on the basic biochemical mechanisms behind drugs of abuse and the etiology and pathogenesis of HIV/AIDS. Based on investigator-generated data, we hypothesize that that Meth and HIV combine to induce changes in miR of DA neurons, specifically miR-9, miR-124, miR-125a, miR-130b, and let-7d; with co- occurring detriments to DA synthesis, metabolism, and transport and these effects can be prevented or reversed by augmenting specific miRs. Centered on this hypothesis, we aim to: 1. Understand the combined effects of Meth and HIV on miR expression, DA synthesis, metabolism, and transport; and 2. Intervene to prevent dopaminergic deficits caused by HIV and Meth by exogenously altering miR function. Our model of study will be cell culture: SH-SY5Y cells and HIV-infected monocytes. Our approach is to use loss-of-function and gain-of-function techniques. By overexpressing or inhibiting specific miRs and measuring dopamine metabolites, transporters, and enzymes, we can understand how miR mediates dopaminergic dysfunction caused by HIV. This project aims to improve HIV treatment and outcomes in drug abusers through a better understanding of the interactions with drugs of abuse and HIV/AIDS disease process. Results from this project will further our understanding of the gene- (regulation through miR) and environment- (HIV-infection) influence risk and protective factors of drug abuse. Knowledge will be applied to help design therapies to reverse or prevent chemical addiction in HIV/AIDS. Results from this project will be leveraged for future research on in understanding the role of miRs in drugs of abuse and HIV/AIDS and medications used to treat HIV. Futures studies also include studying the role of miRs in HIV/AIDS and underlying neurobiology of drug abuse and addiction. PUBLIC HEALTH RELEVANCE: Methamphetamine abuse and human immunodeficiency virus (HIV) infection represent significant public health risks that combine to produce a "double epidemic." This project seeks to understand the molecular genetic intersection and neurochemical changes that occur when neurons are exposed to the two insults. Results from this study would provide a deeper understanding of the molecular mechanisms of neurochemical changes in HIV and methamphetamine abuse and provide possible therapeutic interventions to treat the etiology and neuropathogenesis of HIV in the drug-abusing population.