Mechanisms of blood-brain barrier disruption by an encephalitic virus

脑炎病毒破坏血脑屏障的机制

• 批准号: 8449166
• 负责人: Katherine R. Spindler
• 金额: $ 45.76万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8449166
• 关键词: Actins; Address; Adenoviruses; Astrocytes; Basic Science; Biological Assay; Biological Models; Blood - brain barrier anatomy; Brain; Capsid; Capsid Proteins; Cells; Central Nervous System Diseases; Cleaved cell; Cultured Cells; Cytoskeleton; DNA; DNA Sequence Rearrangement; DNA Viruses; Data; Development; Disease; Drug Delivery Systems; Encephalitis; Endothelial Cells; Family; Fiber; Genetic; Goals; Homeostasis; Human; Immune; Immune response; Immunology; In Situ; In Vitro; Infection; Infectious Agent; Infiltration; Inflammatory; Integrins; Intercellular Junctions; Knock-out; Knowledge; Lead; Leukocytes; Matrix Metalloproteinases; Microglia; Mission; Modeling; Molecular; Molecular and Cellular Biology; Morbidity - disease rate; Mus; Mutant Strains Mice; Natural Immunity; Neurobiology; Outcome; Pathogenesis; Pathway interactions; Pattern; Permeability; Physiological; Play; Proteins; Proteolysis; Public Health; Publishing; RNA Viruses; Research; Resistance; Retroviridae; Role; Signal Pathway; Signal Transduction; System; Testing; Therapeutic Intervention; Tight Junctions; Toxin; United States National Institutes of Health; Vascular Endothelial Cell; Viral; Viral Encephalitis; Viral Genes; Viral Proteins; Virion; Virus; Virus Diseases; Work; base; burden of illness; cell type; chemokine; cytokine; improved; in vivo; in vivo Model; inhibitor/antagonist; innovation; insight; knockout gene; member; mortality; mutant; particle; pathogen; prevent; protein expression; public health relevance; virus host interaction

DESCRIPTION (provided by applicant): Viral encephalitis is a potentially deadly sequela of viral infection for which there are few treatment options. It is frequently associated with blood-brain barrier (BBB) disruption, enabling the entry of virus, inflammatory cells, and deleterious molecules into the brain parenchyma. Members of at least eleven virus families cause encephalitis, including DNA viruses, retroviruses and RNA viruses, with significant morbidity and mortality. Little is known about the mechanisms by which viral infections disrupt the BBB, including the specific viral genes involved and how viruses manipulate host functions that contribute to this important protective barrier. The long-term goal of this research is to improve therapy for people with encephalitis by understanding how viral gene products interact with the host to cause the disease. The overall objective of this application is to determine how a natural mouse pathogen that causes encephalitis, mouse adenovirus type 1 (MAV-1), disrupts the BBB. The central hypothesis is that one or more viral factors induce altered expression and/or function of endothelial cell tight junction proteins, leading to disruption of the BBB. The rationale is that once the mechanism of MAV-1 disruption of the BBB is known, the system can be manipulated genetically and pharmacologically in mice, resulting in innovative approaches for the treatment of encephalitides in humans. The hypothesis will be tested with two specific aims: 1) Identify the primary virus-induced host response leading to altered tight junction protein expression during MAV-1 infection, and 2) Identify the innate signaling and viral components responsible for destruction of BBB integrity. Aim 1 is based on published and preliminary data that MAV-1 reduces tight junction protein expression in brain endothelial cells and increases matrix metalloproteinase (MMP) expression in brains, astrocytes and microglia. Established assays for BBB permeability, MMP activity, and transendothelial resistance will be used to test the hypothesis that MMPs play a role in the decrease in tight junction proteins during infection. Gene knockout-, inhibitor-treated-, and leukocyte-depleted mice will be used to extend these findings in vivo. In Aim 2, viral mutants and physically altered virus particles will be used in in vivo and in vitro infections to test the hypothesis that viral components serve as pathogen-associated molecular patterns that signal innate immunity. To identify which innate immune signaling pathway(s) MAV-1 triggers, leading to BBB disruption, an ordered approach using infection of mouse gene knockouts will be employed. The proposed research is innovative because it uses a comprehensive approach to study BBB disruption caused by a viral pathogen in its natural host, addressing both viral and host contributions to encephalitis. This research will be significant because it will be the first viral mechanism of BBB disruption characterized in vivo in a natural host, contributing an understanding of the action of specific viral gene products and mammalian innate immunity on BBB function. This powerful mouse/MAV-1 model can be manipulated to evaluate innovative approaches to the treatment of encephalitis.

描述(申请人提供)：病毒性脑炎是病毒感染的一种潜在的致命后遗症，治疗方案很少。它经常与血脑屏障(BBB)的破坏有关，使病毒、炎症细胞和有害分子能够进入脑实质。至少有11个病毒家族的成员会引起脑炎，包括DNA病毒、逆转录病毒和RNA病毒，发病率和死亡率都很高。关于病毒感染扰乱血脑屏障的机制，包括涉及的特定病毒基因，以及病毒如何操纵有助于这一重要保护屏障的宿主功能，人们知之甚少。这项研究的长期目标是通过了解病毒基因产品如何与宿主相互作用导致疾病来改善脑炎患者的治疗。这项应用的总体目标是确定一种引起脑炎的自然小鼠病原体-鼠腺病毒1型(MAV-1)如何破坏血脑屏障。中心假设是一个或多个病毒因子诱导内皮细胞紧密连接蛋白的表达和/或功能改变，导致血脑屏障的破坏。其基本原理是，一旦知道MAV-1破坏血脑屏障的机制，就可以在小鼠身上从遗传和药理学上操纵该系统，从而产生治疗人类脑磷脂的创新方法。这一假设将以两个特定的目标进行验证：1)确定病毒诱导的宿主反应，导致MAV-1感染过程中紧密连接蛋白表达的改变；2)确定导致血脑屏障完整性破坏的固有信号和病毒成分。目的1基于已发表的初步数据，MAV-1降低了脑内皮细胞紧密连接蛋白的表达，增加了脑、星形胶质细胞和小胶质细胞的基质金属蛋白酶(MMPs)的表达。已建立的血脑屏障通透性、基质金属蛋白酶活性和跨内皮细胞抵抗力的检测方法将被用来检验MMPs在感染过程中紧密连接蛋白减少中起作用的假设。基因敲除、抑制剂治疗和去白细胞小鼠将被用于在体内扩展这些发现。在目标2中，病毒突变体和物理改变的病毒颗粒将用于体内和体外感染，以检验病毒成分作为病原体相关分子模式信号传递先天免疫的假设。为了确定哪个先天性免疫信号通路(S)MAV-1触发，导致血脑屏障中断，将采用感染小鼠基因敲除的有序方法。这项拟议的研究具有创新性，因为它使用了一种全面的方法来研究病毒病原体在其自然宿主中造成的血脑屏障破坏，解决了病毒和宿主对脑炎的贡献。这项研究将具有重要意义，因为它将是第一个在自然宿主体内表征的BBB破坏的病毒机制，有助于了解特定的病毒基因产物和哺乳动物先天免疫对BBB功能的作用。这种强大的小鼠/MAV-1模型可以被操纵来评估脑炎治疗的创新方法。