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

• 批准号: 8012046
• 负责人: SETH PERRY
• 金额: $ 22.95万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8012046
• 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; 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

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，全球有 3340 万人感染艾滋病毒/艾滋病，新增感染者 270 万人，相关死亡人数 200 万人。 2008年，美国有140万人感染艾滋病毒/艾滋病，新增感染者55,000人，相关死亡人数为25,000人。某些弱势群体或弱势群体承担着不同程度的负担，包括全球的妇女和儿童以及美国黑人 (BA) 和西班牙裔少数群体。 2006 年，尽管 BA 只占总人口的 13%，但其新增感染病例却占 46%。在女性中，BA 承担的负担更加不成比例，2006 年所有新女性 HIV 感染者中 61% 是由 BA 感染的。2006 年男性感染者中，76% 是 MSM。在这些弱势群体中，吸毒（包括安非他明、鸦片制剂、酒精和可卡因）与高危性行为有关，并加速艾滋病毒的进展和死亡率。吸毒也是血清转化的独立危险因素，可预测疾病进展、艾滋病发展和艾滋病死亡率。因此，了解共病药物滥用对 HIV 感染的影响变得越来越重要，特别是因为它影响弱势群体和治疗开发 [1-10]。一些证据表明糖原合酶激酶 3 (GSK3) 可能是外周 HIV 感染的调节因子，并且 GSK3 还可能介导一些滥用药物（例如可卡因和甲基苯丙胺）的作用。然而，对于 GSK3 如何或是否直接影响 HIV 感染（伴或不伴药物滥用）的确切研究还很少，特别是在关键的 HIV 亲和性外周 CD4+ T 细胞和巨噬细胞方面。因此，该提案将研究统一的假设，即 GSK3 激活增加内吞作用，以增强辅助受体介导的外周来源的人类 CD4+ T 细胞和巨噬细胞的 HIV 感染，并且这是可卡因和安非他明增强这些细胞类型的 HIV 感染的机制。首先，我们将确定 GSK3 转基因过度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法进行内吞作用，以及 Rab5 的 siRNA 敲低，我们将确定 GSK3 是否通过增强 Rab5 介导的内吞作用来增加 CD4+ T 细胞和巨噬细胞中的 HIV 感染。在目标 2 中，使用类似的方法，我们将首先确定冰毒或可卡因治疗是否会增加这些细胞类型中 GSK3 的表达和活性以及内吞作用。我们将进一步确定冰毒或可卡因是否会增加这些细胞的 HIV 感染，以及这些影响是否可以通过操纵 GSK3 和/或 Rab5 的表达来消除。在所有这些研究中，无论有或没有可卡因和冰毒，我们将确定 GSK3 抑制化合物锂和 AR-A014418 是否可以改善 GSK3 对 CD4+ T 细胞和巨噬细胞的 HIV 感染的影响，以确定 GSK3 抑制是否是一种有效的治疗方法。减少艾滋病毒病毒载量的潜在可行的治疗策略。因此，这些研究将为调节高度艾滋病毒倾向性外周免疫细胞的艾滋病毒感染的分子机制提供新的见解，以确定治疗合并药物滥用的艾滋病毒疾病的新疗法。 公共健康相关性：该项目探索新的机制理论，可能有助于解释艾滋病毒如何进入人体免疫细胞导致艾滋病毒疾病。更好地了解有关艾滋病毒如何引起艾滋病毒主要细胞类型感染的疾病机制，将有助于确定治疗艾滋病毒和艾滋病的预防性干预的其他治疗靶点。