Effects of HIV-1 Tat protein and methamphetamine on VMAT2-mediated dopamine transmission in the context of neuroHIV and drug abuse

HIV-1 Tat 蛋白和甲基苯丙胺对神经 HIV 和药物滥用背景下 VMAT2 介导的多巴胺传递的影响

• 批准号: 10698618
• 负责人: CHANG-GUO ZHAN
• 金额: $ 65.46万
• 依托单位: UNIVERSITY OF SOUTH CAROLINA AT COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2028-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10698618
• 关键词: AIDS prevention; Acceleration; Acute; Attenuated; Automobile Driving; Behavioral; Binding; Binding Sites; Biological; Biological Assay; Brain; Cell Line; Cells; Chemicals; Cocaine; Cognitive; Cognitive deficits; Computer Models; Development; Dopamine; Doxycycline; Drug abuse; Drug usage; Exhibits; Functional disorder; Gene Expression Profile; Genetic Transcription; Goals; HIV; HIV Infections; HIV Seropositivity; HIV-1; HIV-associated neurocognitive disorder; Homeostasis; Human; Hybrids; Impaired cognition; In Vitro; Incidence; Individual; Infection; Investigation; Knock-in; Knock-in Mouse; Life; Link; Mediating; Methamphetamine; Methamphetamine prevention; Molecular; Mutation; Nervous System Trauma; Neurocognitive Deficit; Nucleus Accumbens; PC12 Cells; Patients; Periodicity; Peripheral; Persons; Play; Potassium; Prefrontal Cortex; Presynaptic Terminals; Prevalence; Proteins; Public Health; Publishing; Rat Transgene; Rewards; Risk; Role; Scanning; Severities; Signal Transduction; Site-Directed Mutagenesis; Slice; Substance abuse problem; Synaptic Vesicles; Therapeutic; Trans-Activators; Transcription Coactivator; Transgenic Mice; Transgenic Organisms; Validation; Vesicle; Viral Proteins; Virus Diseases; Virus Replication; Work; antiretroviral therapy; attenuation; conditioned place preference; dopamine system; dopamine transporter; drug of abuse; extracellular; genetic approach; improved; in vivo; inhibitor; methamphetamine abuse; methamphetamine effect; methamphetamine user; molecular dynamics; mouse model; mutant; neuroAIDS; neuroblastoma cell; neuropsychiatry; novel; pharmacologic; prevent; putamen; transmission process; uptake; vesicular monoamine transporter

PROJECT SUMMARY More than 38 million people are living with Human Immunodeficiency Virus (HIV) infection worldwide which continues to be a global public health problem. Despite the widespread use of efficacious combination of antiretroviral therapy (cART), up to 70% of HIV-positive individuals suffer from cognitive and behavioral deficits collectively known as HIV associated neurocognitive disorders (HAND). Although acute viral replication has been well controlled with cART in the early stage of HIV infection, long-term HIV-1 viral protein exposure within the CNS causes dopaminergic deficits and neurocognitive impairments despite the advent of cART. Substance abuse such as methamphetamine (METH) have been shown to increase the incidence of HAND and exacerbate its severity. Among the viral proteins, HIV-1 transactivator of transcription (Tat) protein plays a significant role in viral replication in the early stage of HIV infection and the pathophysiological effects on development of HAND. Dopamine (DA) transporter (DAT) transports the extracellular dopamine (DA) into cytosolic space of the synaptic terminals, whereas the vesicular monoamine transporter2 (VMAT2) transports the cytosolic DA into synaptic vesicles, whereby both DA transporter and VMAT2 are critical for normal DA homeostasis. Our published studies have demonstrated that Tat protein increases extracellular DA concentration by directly inhibiting DAT, however, the cellular mechanisms underlying Tat-induced inhibition of VMAT2-mediated DA release/uptake remains unexplored. We hypothesize that HIV-1 Tat, acting via the unique binding sites, perturbs the VMAT2 regulatory network that normally sustains concentrative DA transport and potentiates METH’s effect on VMAT2 function, resulting in DA-linked neuropsychiatric dysfunction prominently featured in HAND. Studying single Tat protein allow us to identify targets for Tat binding and develop therapeutic approaches to prevent Tat-mediated neurological damages. The purpose of the proposed investigation is to understand the molecular mechanisms of dysregulation of VMAT2-mediated DA transmission induced by HIV-1 Tat and METH and explore the potential of novel compounds for the prevention of HAND. The specific aims to be pursued in the proposed investigation are: (1) through computational modeling and experimental validation, identify the recognition binding pockets on human VMAT2 for Tat, METH, or novel VMAT2 inhibitors and explore the potential interactions of the inhibitors with Tat and METH, (2) accelerate the pathophysiological roles of VMAT2 in Tat- and METH-dysregulated DA system, and (3) perform proof-of-concept studies using pharmacological and genetic approaches as biological probes to establish their potential for therapeutic application in HAND in concurrent METH users.

项目摘要 全球有超过3800万人患有人类免疫缺陷病毒（HIV）感染 这仍然是全球公共卫生问题。尽管有效地使用了有效组合的宽度 抗逆转录病毒疗法（CART），高达70％的HIV阳性个体患有认知和行为缺陷 统称为HIV相关的神经认知障碍（手）。尽管急性病毒复制具有 在HIV感染的早期，我们通过购物车很好地控制了，长期HIV-1病毒蛋白暴露在内 尽管车冒险冒险，但中枢神经系统会导致多巴胺能防御能力和神经认知障碍。物质 诸如甲基苯丙胺（甲基苯丙胺）之类的滥用已被证明增加了手和 加剧其严重性。在病毒蛋白中，HIV-1转录（TAT）蛋白的反式激活剂播放A 在HIV感染的早期阶段，病毒复制中的重要作用以及病理生理对 手的发展。多巴胺（DA）转运蛋白（DAT）将细胞外多巴胺（DA）传输到 突触末端的胞质空间，而囊泡单胺转运蛋白2（VMAT2）转运 胞质DA进入突触蔬菜，因此DA转运蛋白和VMAT2对于正常DA至关重要 稳态。我们发表的研究表明，TAT蛋白会增加细胞外DA 但是，通过直接抑制DAT浓度，但是，TAT诱导的抑制作用的细胞机制 VMAT2介导的DA释放/吸收仍然出乎意料。我们假设HIV-1 TAT通过 唯一的结合位点，perturbs VMAT2调节网络，该网络通常维持浓缩的DA转运 电势对VMAT2功能的影响，导致DA连接的神经精神功能障碍 突出的手头。研究单TAT蛋白使我们能够识别TAT结合和 开发了预防TAT介导的神经系统损害的理论方法。提议的目的 研究是了解VMAT2介导的DA的失调的分子机制 由HIV-1 TAT和METH诱导的传播，并探索新型化合物预防的潜力 手。在拟议的调查中要追求的具体目的是：（1）通过计算 建模和实验验证，确定人类VMAT2对TAT的识别结合口袋， METH或新型VMAT2抑制剂，并探索抑制剂与TAT和METH的潜在相互作用， （2）加速VMAT2在TAT和METH-DYSCONDED DA系统中的病理生理作用，（3） 使用药物和遗传学方法作为生物学问题进行概念证明研究 在同时发生的甲基甲基使用者中确定他们的治疗应用潜力。