Role of Cell Associated Proteoglycans in HIV-1 Neuropathogenesis

细胞相关蛋白多糖在 HIV-1 神经发病机制中的作用

• 批准号: 7229744
• 负责人: ELIAS G ARGYRIS
• 金额: $ 16.22万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-03-01 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7229744
• 关键词: Affect; Antibodies; Antisense Oligonucleotides; Antisense RNA; Applications Grants; Astrocytes; Biological; Blood - brain barrier anatomy; Brain; CD4 Antigens; Cell Line; Cell surface; Cells; Central Nervous System Infections; Chemicals; Chondroitin ABC Lyase; Chondroitin Sulfate Proteoglycan; Class; Complex; Core Protein; Digestion; Endothelial Cells; Equus caballus; Excision; Family; Gene Family; HIV; HIV Receptors; HIV-1; Heparan Sulfate Proteoglycan; Heparin; Heparitin Sulfate; Human; Immunofluorescence Immunologic; In Vitro; Individual; Infection; K562 Cells; Measurement; Mediating; Messenger RNA; Microglia; Modeling; Molecular; Monoclonal Antibodies; Nerve Degeneration; Neuraxis; Neurons; Neuropathogenesis; Pathway interactions; Patients; Phospholipase C; Play; Proteins; Proteoglycan; RNA; RNA Interference; Relative (related person); Research Personnel; Role; Route; Side; System; T-Lymphocyte; Techniques; Trans-Activators; Viral; Viral Proteins; Virus; Virus Replication; Western Blotting; Work; base; design; glypican 5; heparitinase; improved; inhibitor/antagonist; insight; knock-down; link protein; macrophage; monocyte; monolayer; perlecan; receptor; sodium chlorate; sulfation; syndecan; uptake

DESCRIPTION (provided by applicant): This grant proposal will investigate the role of cell-surface central nervous system (CNS)-specific heparan- sulfate and chondroitin-sulfate proteoglycans (HSPGs and CSPGs respectively) in human immunodeficiency virus type I (HIV-1) neuropathogenesis. The working hypothesis for these studies is that distinct gene- families of cell-associated proteoglycans play a major role in viral capture and transfer via the brain microvascular endothelial cells (MVECs), as we have demonstrated, and other independent investigators have confirmed. In addition, distinct CNS-specific HSPGs may be required for interactions of viral proteins (HIV-1 Tat) with the blood-brain barrier (BBB) and CNS-derived cells. SPECIFIC AIM 1 will investigate the molecular mechanisms by which HIV-1 interacts with MVEC-specific cell-associated proteoglycans to cross the BBB and infect CMS. This will be accomplished by: 1). Complete characterization of MVEC-specific cell- associated proteoglycans in our experimental cellular systems in vitro, 2). Analysis of the role of MVEC- specific proteoglycans in HIV-1 entry and infection of the BBB and CNS by means of alternative approaches, such as chemical, enzymatic, monoclonal antibodies, short interfering RNA (siRNA), and anti-sense. 3). Investigating the role of MVEC-specific proteoglycans in HIV-1 infection of CNS by both free-virus and cell- associated (transmigration of HIV-1 infected monocytes/T-cells through the BBB) virus in our in vitro human BBB model. SPECIFIC AIM 2 will study the molecular mechanisms by which HIV-1-specific proteins (HIV-1 Tat) interact with CNS-specific cell-associated HSPGs to enter and affect CNS-derived cells. By employing similar approaches as in specific AIM1 (i.e. enzymatic, monoclonal antibodies, RNAi, and anti-sense oligonucleotides) we will dissect the participation of specific gene-families of HSPGs (syndecans, glypicans, and/or others) in HIV-1 Tat uptake by human brain MVECs, neurons and astrocytes. In addition, as a model in our studies, we will also employ the human K562-cell line, which lacks HSPGs and CSPGs, and will be transfected with different constructs expressing various human proteoglycans, that are available in our labs. Results from these studies should improve our understanding of the role of CNS-specific proteoglycans in HIV-1 entry and infection of the brain, and may provide insights into strategies for controlling HIV-1 neuroinvasion and associated neurodegeneration in infected patients.

描述（由申请人提供）：该赠款提案将研究细胞表面中枢神经系统（CNS）特异性乙par-硫酸盐和软骨蛋白 - 硫酸软骨蛋白聚糖（HSPGS和CSPGS）在人类免疫缺陷病毒I型（HIV-1）神经疾病中的作用。这些研究的工作假设是，正如我们所证明的那样，与细胞相关的蛋白聚糖的不同基因在病毒捕获和通过脑微血管内皮细胞（MVEC）中起主要作用，并且已经证明了其他独立研究者。此外，病毒蛋白（HIV-1 TAT）与血脑屏障（BBB）和CNS衍生细胞的相互作用可能需要明显的CNS特异性HSPG。具体目标1将研究HIV-1与MVEC特异性细胞相关蛋白聚糖的分子机制，以越过BBB并感染CMS。这将通过：1）完成。在我们的实验性细胞系统中，完全表征MVEC特异性细胞相关的蛋白聚糖，2）。通过替代方法，例如化学，酶促，单克隆抗体，简短的干扰RNA（siRNA）和抗刺激性，分析了MVEC-特异性蛋白聚糖在HIV-1进入HIV-1进入和BBB和CNS感染中的作用。 3）。在我们的体外人类BBB模型中研究了自由病毒和细胞相关的MVEC特异性蛋白聚糖在通过自由病毒和细胞相关的（通过BBB病毒的HIV-1感染单核细胞/T细胞迁移）中的MVEC特异性蛋白聚糖在CNS中的作用。具体目标2将研究HIV-1特异性蛋白（HIV-1 TAT）与CNS特异性细胞相关的HSPG相互作用的分子机制，以进入和影响与CNS衍生的细胞。通过采用与特定目标1（即酶促，单克隆抗体，RNAi和抗势义寡核苷酸）中的类似方法，我们将通过HEV-1 TATES和neuronty neuronty和seerty andy neurons和asty neurons和asty neurons和aster neurons，我们将剖析HSPGS（Syndecans，Glypicans和erse）的特定基因含量的参与。此外，作为我们研究的模型，我们还将采用缺乏HSPG和CSPG的人类K562细胞系，并将用表达各种人类蛋白聚糖的不同构建体转染，这些结构可在我们的实验室中使用。这些研究的结果应提高我们对中枢神经系统特异性蛋白聚糖在HIV-1进入和大脑感染中的作用的理解，并可能提供对控制HIV-1神经浸觉和相关神经变性的策略的见解。