GSK3, endocytosis, and enhanced HIV infection: abused drugs and novel therapies

GSK3、内吞作用和增强的 HIV 感染：滥用药物和新疗法

基本信息

批准号：
8081752
负责人：
SETH PERRY
金额：
$ 18.73万
依托单位：
UNIVERSITY OF ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-01 至 2013-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8081752
关键词：
AIDS neuropathy AIDS/HIV problem Acquired Immunodeficiency Syndrome Actins Adenovirus Vector Affect African American Alcohols Amphetamines Binding Biological Assay Biotinylation Blood Circulation Brain CCR5 gene CD4 Positive T Lymphocytes Cells Cessation of life Child Clathrin Cocaine Conflict (Psychology)Coupling Developing Countries Development Disadvantaged Disease Disease Outcome Disease Progression Dopamine Dopamine D2 Receptor Dopamine Receptor Drug abuse Drug usage Dynamin Endocytosis Female Functional disorder Glycogen Synthase Kinase 3 HIV HIV Infections HIV therapy Hispanics Human Immune Immune system Infection Intervention Investigation Label Lead Learning Life Link Lithium Lithium Compounds Mediating Mediator of activation protein Membrane Methamphetamine Minority Mitotic Molecular Molecular Biology Monitor Opiates Oxidative Stress Pathology Peripheral Pharmaceutical Preparations Pharmacologic Actions Play Population Proteins Regulation Relative (related person)Replication-Associated Process Reporting Risk Risk Factors Role Running Signal Transduction Small Interfering RNA Techniques Testing Therapeutic Time Transgenes Ursidae Family Viral Load result Vulnerable Populations Western Blotting Woman Work cell type cellular transduction dopamine transporter drug of abuse forgetting global health gp160 high risk sexual behavior inhibitor/antagonist insight macrophage male meetings mortality novel novel therapeutic intervention overexpression pandemic disease prevent public health relevance receptor research study success theories therapeutic target therapy design therapy development vaccine development valproate vector

项目摘要

DESCRIPTION (provided by applicant): Worldwide 33.4 million people lived with HIV/AIDS in 2008, with 2.7 million new infections, and 2 million related deaths. In the US, 1.4 million people had HIV/AIDS in 2008, with 55,000 new infections, and 25,000 related deaths. Certain disadvantaged or vulnerable populations bear a disparate share of this burden, including women and children globally, and Black American (BA) and Hispanic minorities in USA. BAs acquired 46% of new infections in 2006, despite comprising only 13% of the population. Among females, BAs share an even more disproportionate burden, acquiring 61% of all new female HIV infections in 2006. For 2006 male infections, 76% were MSM. In these vulnerable groups, drug use including amphetamines, opiates, alcohol, and cocaine, is associated with high-risk sexual behavior, and accelerates progression of and mortality from HIV. Drug use is also an independent risk factor for seroconversion, and predictive of disease progression, development of AIDS, and AIDS mortality. Thus it is increasingly important to understand impacts of comorbid drug abuse on HIV infection, particularly as it affects vulnerable populations and therapy development [1-10]. Some evidence implicates glycogen synthase kinase 3 (GSK3) as a possible regulator of peripheral HIV infection, and GSK3 also may mediate effects of some abused drugs, such as cocaine and methamphetamine. However there has been little investigation of exactly how or whether GSK3 directly affects HIV infection with and without comorbid drug abuse, particularly as regards key HIV tropic peripheral CD4+ T cells and macrophages. Hence, this proposal will investigate the unifying hypothesis that GSK3 activation increases endocytosis to enhance coreceptor mediated HIV infection of peripherally derived human CD4+ T cells and macrophages, and that this is the mechanism by which cocaine and amphetamines enhance HIV infection of these cell types. First we will determine if GSK3 transgene overexpression or knockdown directly affects HIV infection of CD4+ T cells and macrophages. Next, using fluorescent, immunocytochemical, and western blot assays for endocytosis, and siRNA knockdown of Rab5, we will determine whether GSK3 increases in HIV infection in CD4+ T cells and macrophages by enhancing Rab5-mediated endocytosis. In Aim 2, using similar approaches, we will first determine whether Meth or cocaine treatment increases GSK3 expression and activity, and endocytosis, in these cell types. We will further determine whether Meth or cocaine increase HIV infection of these cells, and whether these effects can be abrogated by manipulating GSK3 and/or Rab5 expression. In all of these studies, with and without cocaine and Meth, we will determine whether GSK3's effects on HIV infection of CD4+ T cells and macrophages can be ameliorated by the GSK3-inhibiting compounds lithium and AR-A014418, to determine whether GSK3 inhibition is a potentially viable therapeutic strategy for reducing viral load in HIV disease. Thus these studies will provide new insights into the molecular mechanisms regulating HIV infection of highly HIV-tropic peripheral immune cells, to identify new therapies for treating HIV disease with comorbid drug abuse. PUBLIC HEALTH RELEVANCE: This project explores new mechanistic theories that may help explain how HIV enters human immune cells to cause HIV disease. Better understanding of these disease mechanisms concerning how HIV causes infection of principal cell types involved in HIV, will help identify additional therapeutic targets for preventative intervention to treat HIV and AIDS.

描述（由申请人提供）：2008年全球3340万人患有艾滋病毒/艾滋病，有270万新的感染和200万相关死亡。在美国，2008年有140万人患有艾滋病毒/艾滋病，有55,000人新感染和25,000例相关死亡。某些处境不利或脆弱的人群在这一负担中占有不同的份额，包括全球妇女和儿童，黑人美国人（BA）和美国的西班牙裔少数民族。尽管仅占人口的13％，但BAS在2006年获得了46％的新感染。在女性中，BAS的负担更加不成比例，在2006年获得了所有新女性HIV感染的61％。对于2006年男性感染，MSM为76％。在这些脆弱的群体中，包括苯丙胺，阿片类药物，酒精和可卡因在内的药物使用与高风险的性行为有关，并加速了HIV的进展和死亡率。药物使用也是血清转化的独立危险因素，可以预测疾病进展，艾滋病的发展和艾滋病死亡率。因此，了解合并药物滥用对艾滋病毒感染的影响越来越重要，尤其是因为它影响脆弱的人群和治疗发展[1-10]。一些证据表明，糖原合酶激酶3（GSK3）可能是外周HIV感染的可能调节剂，而GSK3也可能介导某些滥用药物的作用，例如可卡因和甲基苯丙胺。然而，几乎没有研究GSK3如何或如何直接影响滥用药物的艾滋病毒感染，尤其是在关键的HIV热带外围CD4+ T细胞和巨噬细胞的情况下。因此，该提案将研究以下统一的假设：GSK3激活会增加内吞作用，以增强对外周衍生的人CD4+ T细胞和巨噬细胞的介导的介导的HIV感染，这是可卡因和Amphetamine增强这些细胞型艾滋病毒感染的机制。首先，我们将确定GSK3转基因过表达或敲低是否直接影响CD4+ T细胞和巨噬细胞的HIV感染。接下来，使用荧光，免疫细胞化学和蛋白质印迹分析进行内吞作用，以及RAB5的siRNA敲低，我们将通过增强RAB5介导的内吞作用来确定GSK3是否会增加CD4+ T细胞和巨噬细胞中HIV感染的HIV感染。在AIM 2中，使用类似的方法，我们将首先确定在这些细胞类型中，甲基苯甲酸酯或可卡因治疗是否会增加GSK3的表达和活性以及内吞作用。我们将进一步确定甲基苯细胞或可卡因是否会增加这些细胞的HIV感染，以及是否可以通过操纵GSK3和/或RAB5表达来消除这些作用。在所有这些研究中，有或没有可卡因和甲基甲基苯丙胺，我们将确定GSK3对CD4+ T细胞和巨噬细胞的HIV感染的影响是否可以通过抑制GSK3化合物锂和AR-A014418来改善，以确定GSK3抑制是否可能可行的治疗策略。因此，这些研究将提供有关调节高度HIV - 热带周围免疫细胞HIV感染的分子机制的新见解，以鉴定与合并药物滥用治疗HIV疾病的新疗法。公共卫生相关性：该项目探讨了新的机械理论，可以帮助解释艾滋病毒如何进入人类免疫细胞引起艾滋病毒疾病。更好地理解有关HIV如何引起HIV中主要细胞类型感染的这些疾病机制，将有助于确定其他治疗靶标的预防性干预措施治疗HIV和AIDS。