P2: MOLECULAR MECHANISMS OF WEST NILE VIRUS NEUROINVASION

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 西尼罗河病毒（WNV）是由蚊子传播的，通常会导致人类轻微疾病。然而，在大约2%的感染者中，西尼罗河病毒会导致致命的脑部疾病，称为脑膜脑炎。目前，没有治疗西尼罗河病毒的药物或预防人类感染西尼罗河病毒的疫苗。我们的初步研究表明，西尼罗河病毒通过穿过血脑屏障（BBB）进入大脑，并产生介质，如称为基质金属蛋白酶（MMPs）的酶蛋白，降解BBB。我们的中心假设是，WNV感染的血液和脑细胞通过环氧合酶-2（考克斯-2）信号通路增加MMPs的产生，通过降解特异性紧密连接蛋白（TJP）介导BBB破坏。 我们将追求三个具体目标：目标1将分析考克斯-2酶及其产物前列腺素E2在调节MMPs水平中的作用。在目标2中，我们将评估WNV感染的单核细胞降解TJP的能力及其穿过良好建立的体外BBB模型的迁移。目的3将采用小鼠模型来表征WNV诱导的TJP改变。此外，我们还将确定MMP抑制剂预防BBB破坏和WNV最终进入CNS的潜力。我们使用细胞培养和动物模型的综合研究将突出MMPs诱导的BBB破坏在WNV疾病发病机制中的作用。对MMP和TJP在BBB破坏中的功能以及MMP抑制剂改善WNV疾病结果的能力的新见解的发现将具有相当大的治疗相关性。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. West Nile virus (WNV), which is transmitted by mosquitoes, usually causes a mild disease in humans. In approximately 2% of infected people, however, WNV causes a fatal brain disease, called meningoencephalitis. Currently, there are no drugs to treat WNV or vaccines to prevent WNV infection in humans. Our preliminary studies indicate that WNV gains entry into the brain by crossing the blood-brain barrier (BBB) and produces mediators, such as enzymatic proteins called matrix metalloproteinases (MMPs), that degrade the BBB. Our central hypothesis is that the increased production of MMPs by WNV-infected blood and brain cells, via the cyclooxygenase-2 (COX-2) signaling pathway, mediates BBB breakdown by degrading specific tight junction proteins (TJP). We will pursue three specific aims: Aim 1 will analyze the role of COX-2 enzyme and its product, Prostaglandin E2, in regulating the levels of MMPs. In Aim 2, we will assess the ability of WNV-infected monocytes to degrade TJP and its transmigration across a well-established in vitro BBB model. Aim 3 will employ a mouse model to characterize WNV-induced alterations in TJP. Further, we will also determine the potential of MMP inhibitors to prevent BBB disruption and eventual entry of WNV into the CNS. Our comprehensive study using cell culture and animal model will highlight the role of MMPs-induced BBB disruption in WNV disease pathogenesis. The discovery of new insights into the function(s) of MMPs and TJP in BBB disruption and ability of MMP inhibitors to improve WNV disease outcome would have considerable therapeutic relevance.