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：
8254417
负责人：
Anuja Ghorpade
金额：
$ 39.3万
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/8254417
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

ABSTRACT: 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 exitotoxic 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.

抽象的：甲基苯丙胺（甲基苯丙胺），一种成瘾性刺激剂对中枢神经系统具有持久的毒性作用（CNS）。临床研究表明，甲基依赖性对神经心理缺陷具有加性作用与HIV-1感染相关。氧化应激，兴奋性毒性，BBB损伤和神经胶质细胞激活，所有这些已经与甲基和HIV-1相关的神经毒性的机制独立涉及。这提案将调查HIV-1 CNS感染和滥用MET的特定机制，导致氧化应激和NF-KB信号的总体增加，导致星形胶质细胞损害和内皮细胞功能。我们建议甲基介导的星形胶质细胞中的氧化应激导致下降调节出口毒性氨基酸转运蛋白（EAAT）-2，主要星形胶质细胞谷氨酸清除剂，而在内皮细胞，氧化应激的增加会导致BBB完整性的丧失。确实，我们的初步数据表明，甲基甲基暴露导致星形胶质细胞和人类原发性BMVEC的ROS产生，在星形胶质细胞中，EAAT-2表达降低，在体外降低了BBB完整性，抗氧化剂恢复处理，导致紧密连接蛋白的表达/重新分布，粘附增强和单核细胞在内皮单层中的迁移和BMVEC中激活的小GTPase的迁移以前与HIV-1脑炎（HIVE）期间的BBB损伤有关。展示了新的初步数据 BBB在体内的渗透性增加了暴露于抗氧化剂阻止的甲基的动物治疗。该建议将研究一种新的机械通用性概念，即甲基介导的氧化应激在星形胶质细胞和内皮细胞加剧损伤中发挥细胞特异性作用由HIV-1 CNS感染和使用体内的这些靶标划定治疗选择引起的研究。结合体外测定和甲基动物动物模型，我们将解决以下内容问题：增强的活性氧，ROS产生和NF-KB信号的作用是什么 EAAT-2在星形胶质细胞中的表达/功能减少？（AIM 1）BBB的基本机制是什么功能障碍和增强的单核细胞粘附/跨脑内皮的迁移（通过氧化应激，干扰NF-KB和GTPase信号传导）？（目标2）;并且可以减少氧化应激的治疗剂正常化的EAAT-2功能可以改善BBB功能障碍和神经毒性的动物模型和滥用甲基苯丙胺？（目标3）我们将利用原发性人星形胶质细胞和大脑解决这些问题微血管细胞，体外BBB模型，并评估甲基和HIV-1相关刺激对对 EAAT-2和BBB功能。抗氧化剂和特定信号抑制剂将用于描述途径参与了这些效果。我们的蜂巢动物模型将用于研究这些细胞特异性机制在认知功能，BBB损伤和神经毒性中。我们相信拟议的作品非常重要，因为它们将发现涉及组合效应的新型机制根据这些研究，在中枢神经系统中的HIV-1和METH的治疗方法提出了治疗方法。