Mechanisms and Interventions for Methamphetamine and HIV-1 Induced CNS Injury

甲基苯丙胺和 HIV-1 引起的中枢神经系统损伤的机制和干预措施

基本信息

批准号：
8051739
负责人：
Anuja Ghorpade
金额：
$ 39.5万
依托单位：
TEMPLE UNIV OF THE COMMONWEALTH
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-04-01 至 2014-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8051739
关键词：
3-nitrotyrosine Address Adhesions Agonist American Amino Acid Transporter Animal Model Animals Anti-Inflammatory Agents Anti-inflammatory Antioxidants Area Astrocytes Behavior Biological Biological Assay Blood - brain barrier anatomy Brain CCR5 gene CXCR4 gene Cell Adhesion Molecules Cell Survival Cell physiology Cells Central Nervous System Infections Cessation of life Chronic Clinical Research Cocaine Cognitive deficits Consultations Cytoskeleton Data Dementia Detection Down-Regulation Drug usage Electrical Resistance Encephalitis Endothelial Cells Endothelium Excitatory Amino Acid Transporter 2 Functional disorder Future Generations Genetic Transcription Glutamate Transporter Glutamates Guanosine Triphosphate Phosphohydrolases HIV HIV-1 Health Human Immune Immune response Impaired cognition Impairment In Vitro Individual Infection Inflammation Mediators Injury Intervention Investigation Label Lead Leukocytes Levocarnitine Acetyl Link Magnetic Resonance Imaging Measures Mediating Methamphetamine Methamphetamine dependence Modeling Molecular Weight Monomeric GTP-Binding Proteins Mononuclear Mus Myosin Light Chain Kinase NF-kappa B NOD/SCID mouse Nerve Nerve Degeneration Neuraxis Neuroglia Neuronal Injury Neurons Non obese Outcome Oxidative Stress PPAR gamma Pathogenesis Pathway interactions Peripheral Blood Lymphocyte Permeability Pharmaceutical Preparations Phosphorylation Play Production Property Proteins Reactive Oxygen Species Reader Reading Regulation Rewards Risk Role Scheme Secondary to Signal Transduction Specimen Spinal Ganglia Stimulus Surveys Testing Therapeutic Therapeutic Intervention Tight Junctions Time Toxic effect Tracer Transcriptional Activation Trolox Tyrosine 3-Monooxygenase Viral Vitamin E Water Western Blotting Work adipocyte differentiation analog astrogliosis base brain tissue central nervous system injury chemokine cognitive function diabetic excitotoxicity extracellular functional disability immunocytochemistry in vitro Assay in vivo inhibitor/antagonist insight insulin sensitivity mRNA Expression macrophage methamphetamine abuse methamphetamine exposure methylxanthine migration monocyte monolayer nerve injury neuroinflammation neuropsychological neurotoxic neurotoxicity novel prevent propentofylline protective effect protein expression reconstitution research study response stimulant abuse therapeutic target transcription factor uptake

项目摘要

DESCRIPTION (provided by applicant): Methamphetamine (METH), an addictive stimulant has long lasting toxic effects on the central nervous system (CNS). Clinical studies indicated that METH dependence has an additive effect on neuropsychological deficits associated with HIV-1 infection. Oxidative stress, excitotoxicity, BBB impairment and glial cell activation, all have been independently implicated in the mechanisms of METH- and HIV-1-associated neurotoxicity. This proposal will investigate specific mechanisms operative in HIV-1 CNS infection and METH abuse that lead to an overall increase in oxidative stress and NF-kB signaling resulting in impairment of astrocytes and endothelial cell function. We propose that METH-mediated oxidative stress in astrocytes leads to a down regulation excitotoxic amino acid transporter (EAAT) -2, the primary astrocyte glutamate scavenger, while in endothelial cells, such an increase in oxidative stress results in loss of BBB integrity. Indeed, our preliminary data suggest that METH exposure caused ROS generation in astrocytes and human primary BMVEC, diminished EAAT-2 expression, in astrocytes, decreased BBB integrity in vitro that was restored by antioxidant treatment, caused decreased expression/re-distribution of tight junction proteins, enhanced adhesion and migration of monocytes across endothelial monolayers and activated small GTPases in BMVEC that were previously implicated in the BBB injury during HIV-1 encephalitis (HIVE). New preliminary data demonstrated increased permeability of BBB in vivo in animals exposed to METH that was prevented by antioxidant treatment. This proposal will investigate a novel concept of mechanistic commonality, i.e. METH-mediated oxidative stress exerts its cell-specific effects in astrocytes and endothelial cells aggravating injury caused by HIV-1 CNS infection and delineate therapeutic options for these targets using in vivo studies. Using a combination of in vitro assays and METH/HIVE animal model, we will address the following questions: What is the role(s) of enhanced reactive oxygen species, ROS production and NF-kB signaling in diminished expression/function of EAAT-2 in astrocytes? (Aim 1) What are underlying mechanisms of BBB dysfunction and enhanced monocyte adhesion/migration across brain endothelium (via oxidative stress, interference with NF-kB and GTPase signaling)? (Aim 2); and Can therapeutics decreasing oxidative stress and normalizing EAAT-2 function ameliorate BBB dysfunction and neurotoxicity in an animal model for HIVE and METH abuse? (Aim 3) We will address these questions utilizing primary human astrocytes and brain microvascular cells, in vitro BBB models and evaluate combined effects of METH and HIV-1 relevant stimuli on EAAT-2 and BBB function. Antioxidants and specific signaling inhibitors will be utilized to delineate pathways involved in these effects. Our HIVE animal model will be employed to investigate the biological outcomes of these cell-specific mechanisms in cognitive function, BBB damage and neurotoxicity. We believe that the proposed works are highly significant, as they will uncover novel mechanisms involved in the combined effects of HIV-1 and METH in the CNS and propose therapeutic approaches based on these investigations. PUBLIC HEALTH RELEVANCE: Methamphetamine, a highly addictive stimulant abused by millions of Americans has long lasting toxic effects on the central nervous system. Clinical studies indicated that methamphetamine dependence has an addictive effect on cognitive deficits associated with HIV-1 infection. Current proposal aims to understand mechanisms of combined effects of HIV-1 and methamphetamine in the brain and to propose neuroprotective therapies.

描述（由申请人提供）：甲基苯丙胺（METH）是一种成瘾兴奋剂，对中枢神经系统（CNS）具有长期持续的毒性作用。临床研究表明，METH 依赖对与 HIV-1 感染相关的神经心理缺陷具有累加效应。氧化应激、兴奋性毒性、血脑屏障损伤和神经胶质细胞活化，所有这些都独立地涉及 METH 和 HIV-1 相关神经毒性的机制。该提案将研究 HIV-1 CNS 感染和 METH 滥用中的具体机制，这些机制会导致氧化应激和 NF-kB 信号传导全面增加，从而导致星形胶质细胞和内皮细胞功能受损。我们提出，星形胶质细胞中冰毒介导的氧化应激会导致兴奋性毒性氨基酸转运蛋白（EAAT）-2（主要的星形胶质细胞谷氨酸清除剂）下调，而在内皮细胞中，氧化应激的增加会导致血脑屏障完整性丧失。事实上，我们的初步数据表明，METH 暴露导致星形胶质细胞和人原代 BMVEC 中 ROS 产生，星形胶质细胞中 EAAT-2 表达减少，体外 BBB 完整性降低（通过抗氧化治疗恢复），导致紧密连接表达减少/重新分布蛋白，增强单核细胞跨内皮单层的粘附和迁移，并激活 BMVEC 中的小 GTP 酶，这些酶先前与 HIV-1 脑炎 (HIVE) 期间的 BBB 损伤有关。新的初步数据表明，接触冰毒的动物体内血脑屏障的通透性增加，而抗氧化治疗可以防止这种情况发生。该提案将研究机制共性的新概念，即冰毒介导的氧化应激在星形胶质细胞和内皮细胞中发挥其细胞特异性作用，从而加重 HIV-1 CNS 感染引起的损伤，并利用体内研究为这些靶标制定治疗方案。结合体外测定和 METH/HIVE 动物模型，我们将解决以下问题：增强的活性氧、ROS 产生和 NF-kB 信号传导在 EAAT-2 表达/功能减弱中的作用是什么在星形胶质细胞中？（目标 1）BBB 功能障碍和单核细胞跨脑内皮粘附/迁移增强（通过氧化应激、干扰 NF-kB 和 GTPase 信号传导）的潜在机制是什么？（目标 2）；减少氧化应激和使 EAAT-2 功能正常化的疗法能否改善 HIVE 和 METH 滥用动物模型中的 BBB 功能障碍和神经毒性？（目标 3）我们将利用原代人星形胶质细胞和脑微血管细胞、体外 BBB 模型来解决这些问题，并评估 METH 和 HIV-1 相关刺激对 EAAT-2 和 BBB 功能的综合影响。抗氧化剂和特定信号抑制剂将用于描绘这些效应所涉及的途径。我们的 HIVE 动物模型将用于研究这些细胞特异性机制在认知功能、血脑屏障损伤和神经毒性方面的生物学结果。我们相信，拟议的工作非常重要，因为它们将揭示 HIV-1 和 METH 在中枢神经系统中联合作用的新机制，并根据这些研究提出治疗方法。公共卫生相关性：甲基苯丙胺是一种高度成瘾的兴奋剂，被数百万美国人滥用，对中枢神经系统具有长期持续的毒性作用。临床研究表明，甲基苯丙胺依赖对与 HIV-1 感染相关的认知缺陷具有成瘾作用。目前的提案旨在了解 HIV-1 和甲基苯丙胺在大脑中联合作用的机制，并提出神经保护疗法。