HIV-Induced Dopamnergic Changes and Methamphetamine Toxicity Mediated through Mic

描述（由申请人提供）：这是一个试点项目，以回应PAR-10-021在药物滥用研究艾滋病毒/艾滋病的广泛领域。该项目的目标是了解HIV感染的巨噬细胞如何与甲基苯丙胺（Methe）结合联合收割机，以引起多巴胺能神经细胞的改变，并确定靶向microRN（miR）是否可以逆转或预防功能障碍。滥用的药物，如甲基苯丙胺和可卡因，在化学上与多巴胺相似，通过逆转多巴胺转运蛋白从而激活多巴胺受体并导致高浓度多巴胺在突触空间中积聚而引起大脑中的多巴胺能功能障碍。此外，甲基苯丙胺引起多巴胺能神经元中囊泡单胺转运蛋白的功能障碍，导致囊泡多巴胺的包装功能障碍、多巴胺在细胞质中的积累和下游氧化应激。众所周知，HIV和病毒感染会导致长期感染的大脑中多巴胺能细胞的损失，尽管其机制尚不清楚。艾滋病毒感染人群中的药物滥用和药物成瘾构成了重大的公共卫生风险，并使人们对药物滥用背后的基本生物化学机制以及艾滋病毒/艾滋病的病因和发病机制有了新的认识。基于代谢仪产生的数据，我们假设甲基苯丙胺和HIV联合收割机诱导DA神经元的miR变化，特别是miR-9、miR-124、miR-125 a、miR-130 b和let-7 d;同时发生对DA合成、代谢和转运的损害，并且这些作用可以通过增加特异性miR来预防或逆转。围绕这一假设，我们的目标是：1。了解甲基苯丙胺和HIV对miR表达、DA合成、代谢和转运的联合作用; 2.通过外源性改变miR功能进行干预，以预防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-SY 5 Y细胞和HIV感染的单核细胞。我们的方法是使用功能丧失和功能获得技术。通过过度表达或抑制特定的miR和测量多巴胺代谢物，转运蛋白和酶，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药物滥用与艾滋病毒/艾滋病疾病过程的相互作用，改善药物滥用者的艾滋病毒治疗和成果。该项目的结果将进一步加深我们对基因（通过miR调节）和环境（HIV感染）影响药物滥用风险和保护因素的理解。知识将用于帮助设计治疗方法，以扭转或预防艾滋病毒/艾滋病的化学品成瘾。该项目的结果将用于未来的研究，以了解miRs在滥用药物和艾滋病毒/艾滋病以及用于治疗艾滋病毒的药物中的作用。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：甲基安非他明滥用和人体免疫缺陷病毒（艾滋病毒）感染是重大的公共卫生风险，联合收割机结合在一起，产生了“双重流行病”。“这个项目旨在了解当神经元暴露于两种侮辱时发生的分子遗传交叉和神经化学变化。这项研究的结果将提供更深入的了解艾滋病毒和甲基苯丙胺滥用的神经化学变化的分子机制，并提供可能的治疗干预措施，以治疗吸毒人群中艾滋病毒的病因和神经发病机制。

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.