Enterovirus entry into polarized endothelial cells

肠道病毒进入极化内皮细胞

• 批准号: 8295004
• 负责人: Carolyn B Coyne
• 金额: $ 35.61万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2013-09-05
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8295004
• 关键词: Accounting; Actins; Acute Hemorrhagic Conjunctivitis; Adult; Antiviral Agents; Aseptic Meningitis; Biochemical; Biological; Biological Assay; Blood - brain barrier anatomy; Brain; Bypass; Candidate Disease Gene; Cell Line; Cell physiology; Cells; Child; Complex; Coupled; Coxsackie B Viruses; Coxsackie Viruses; Cytoskeleton; Diabetes Mellitus; Dominant-Negative Mutation; Echo Viruses; Endocytosis; Endothelial Cells; Endothelium; Enterovirus; Enterovirus 71; Enterovirus Infections; Epithelial; Epithelial Cells; Epithelium; Fever; Fluorescence; Gastrointestinal tract structure; Genes; Genome; Health; Human; Human poliovirus; Infection; Integration Host Factors; Intestines; Lead; Libraries; Mediating; Modeling; Myocarditis; Neuraxis; Oral; Pathway interactions; Pericarditis; Phosphotransferases; Play; Polioviruses; Process; Protein Tyrosine Kinase; RNA Interference; Role; Route; Series; Signal Transduction; Signaling Molecule; Site; Structure; Surface; Syndrome; TEC Protein Tyrosine Kinase; Techniques; Therapeutic; United States; Viral; Virus; base; cell type; combat; comparative; design; human TNK1 protein; inhibitor/antagonist; insight; intestinal epithelium; monolayer; pathogen; polarized cell; prevent; research study; secondary infection

DESCRIPTION (provided by applicant): The mucosal surface is the site of entry of >90% of common infectious pathogens in humans. However, studies of virus entry and infection are largely limited to non-polarized cells and little is known regarding the mechanisms used by viral pathogens to infect polarized cell monolayers. Enteroviruses are ideal pathogens for the study of polarized cell-virus interactions as they have evolved highly efficient strategies to bypass both epithelial and endothelial barriers for infection. Our previous studies have established that coxsackievirus B (CVB) and poliovirus (PV) enter polarized cells by endocytic mechanisms that require activation of specific intracellular signaling molecules that drive actin cytoskeleton reorganization, junctional complex modulation, and eventual virus endocytosis. Our preliminary studies suggest that CVB endocytosis into polarized intestinal epithelia is distinct from that in polarized BBB endothelia and that the signaling molecules required to facilitate virus entry are disparate between these cell types. Based on these observations, we hypothesize that enterovirus entry into polarized epithelial and endothelial cells is mediated by cell-type-specific intracellular signals and occur through different endocytic mechanisms. In this proposal, we have designed experiments to more clearly define the cellular pathways hijacked by CVB to gain entry into polarized human brain microvascular endothelial cells (HBMEC), a model of the blood-brain barrier. Specifically, we will (1) define the endocytic pathway used by CVB to gain entry into HBMEC (Specific Aim 1), (2) characterize the role of intracellular kinases in facilitating CVB entry into HBMEC (Specific Aim 2), and (3) compare the intracellular signaling molecules required for enterovirus infection in a variety of cell types (Specific Aim 3). Given the diverse pathological complications associated with enterovirus infections, these studies will provide insights into how enteroviruses have evolved to infect polarized cells, and will serve as a model for how other viral pathogens circumvent the barriers presented by polarized cell monolayers. Furthermore, as there are currently no effective therapeutics to treat enterovirus infections, these studies may lead to the design of cell-type-specific anti-viral targets. PUBLIC HEALTH RELEVANCE: Enteroviruses (which include coxsackievirus, poliovirus, and echovirus) account for as many as 15 million symptomatic infections in the United States each year. There are currently no effective therapeutics to combat enteroviral infections. The studies in this proposal will identify cellular genes required for mediating enterovirus infection and may lead to the design of cell-type-specific antiviral targets.

描述（由申请人提供）：粘膜表面是人类 90% 以上的常见传染性病原体的进入部位。然而，对病毒进入和感染的研究很大程度上局限于非极化细胞，并且对于病毒病原体感染极化细胞单层的机制知之甚少。肠道病毒是研究极化细胞-病毒相互作用的理想病原体，因为它们已经进化出高效的策略来绕过上皮和内皮屏障进行感染。我们之前的研究已经确定，柯萨奇病毒 B (CVB) 和脊髓灰质炎病毒 (PV) 通过内吞机制进入极化细胞，该机制需要激活特定的细胞内信号分子，驱动肌动蛋白细胞骨架重组、连接复合物调节和最终的病毒内吞作用。我们的初步研究表明，极化肠上皮细胞中的 CVB 内吞作用与极化 BBB 内皮细胞中的内吞作用不同，并且促进病毒进入所需的信号分子在这些细胞类型之间是不同的。基于这些观察，我们假设肠道病毒进入极化上皮和内皮细胞是由细胞类型特异性的细胞内信号介导的，并通过不同的内吞机制发生。在这项提案中，我们设计了实验，以更清楚地定义 CVB 劫持进入极化人脑微血管内皮细胞 (HBMEC)（血脑屏障模型）的细胞途径。具体来说，我们将 (1) 定义 CVB 用于进入 HBMEC 的内吞途径（具体目标 1），(2) 描述细胞内激酶在促进 CVB 进入 HBMEC 中的作用（具体目标 2），以及 (3)比较多种细胞类型中肠道病毒感染所需的细胞内信号分子（具体目标 3）。鉴于与肠道病毒感染相关的多种病理并发症，这些研究将深入了解肠道病毒如何进化以感染极化细胞，并将作为其他病毒病原体如何绕过极化细胞单层所呈现的障碍的模型。此外，由于目前没有有效的疗法来治疗肠道病毒感染，这些研究可能会导致细胞类型特异性抗病毒靶点的设计。公共卫生相关性：在美国，肠道病毒（包括柯萨奇病毒、脊髓灰质炎病毒和埃可病毒）每年导致多达 1500 万例有症状感染。目前尚无有效的疗法来对抗肠道病毒感染。该提案中的研究将确定介导肠道病毒感染所需的细胞基因，并可能导致细胞类型特异性抗病毒靶标的设计。