Elucidating binding, fusion and entry of HMPV in cell culture and host tissue

阐明 HMPV 在细胞培养物和宿主组织中的结合、融合和进入

• 批准号: 8908750
• 负责人: Edita Maria Klimyte
• 金额: $ 3.29万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-05-01 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8908750
• 关键词: A549; Address; Adult; Affect; Antigens; Antiviral Agents; Area; Asthma; Binding; Biological Models; Bronchiolitis; Caveolins; Cell Culture Techniques; Cell Line; Cell membrane; Cells; Chemicals; Child; Childhood; Chimeric Proteins; Chronic; Chronic Disease; Clathrin; Clinical; Collection; Complex; Dendrimers; Dependence; Development; Disease; Dynamin; Elderly; Endocytosis; Endosomes; Epithelial; Epithelium; Escherichia coli; Excision; Family; Glycoproteins; HIV; Heparan Sulfate Proteoglycan; Heparitin Sulfate; Hospitalization; Human; Human Metapneumovirus; Human respiratory syncytial virus; Immunocompromised Host; Infant; Infection; Influenza; Inorganic Sulfates; Intervention; Label; Laboratories; Life Cycle Stages; Lipids; Lung; Lung diseases; Mediating; Membrane Fusion; Modeling; Molecular; Mucous body substance; Paramyxoviridae; Paramyxovirus; Pathway interactions; Patients; Peptides; Play; Pneumonia; Pneumovirinae; Population; Predisposition; Production; RNA Viruses; Respiratory Tract Diseases; Respiratory syncytial virus; Role; Route; Site; Structure of parenchyma of lung; Surface; Symptoms; System; Time; Tissue Model; Tissues; Trypsin; Unspecified or Sulfate Ion Sulfates; Vaccines; Viral; Virus; airway epithelium; airway hyperresponsiveness; cell type; cellular targeting; clinically significant; in vivo; inhibitor/antagonist; member; monolayer; mortality; novel; particle; pathogen; public health relevance; receptor; reconstitution; respiratory; syndecan; uptake

 DESCRIPTION (provided by applicant): Human metapneumovirus (HMPV) is a recently discovered respiratory pathogen in the Paramyxoviridae family that infects nearly 100% of the world population. This enveloped virus causes severe viral respiratory disease in infants, the elderly, and immunocompromised patients worldwide. In fact HMPV is estimated to be the second most common cause of pediatric lower respiratory illness, following the closely related respiratory syncytial virus. Furthermore, the rate of hospitalization for respiratory disease caused by HMPV in adults over the age of 65 is near that of influenza. Despite its clinical significance there is no antiviral treatment or vaccine available at this time. Since its discovery several key factors have been identified to play a role in HMPV entry, but much of the mechanisms of binding, fusion, and entry remain unsolved. It has been previously shown by our group that heparan sulfate is a required moiety for HMPV binding and infectivity; however, it is not known if heparan sulfate proteoglycans (HSPGs) serve as a receptor or mediate other necessary cellular factors on the plasma membrane. Furthermore, the route of entry has not been identified for HMPV. Thus, to elucidate the role of HSPGs, we will utilize a collection of currently available HMPV strains from laboratory strains and clinical isolates to determine the effects of heparan sulfate blocking molecules and knockdown studies on specific HSPGs; in order to identify the route of entry, we will utilize viral particles with fluorescent labeled envelopes to track if membrane fusion is occurring at the plasma membrane or within intracellular compartments. Finally we propose to study HMPV entry in the context of traditional cell culture, utilizing physiologically relevant cell lines derived from human lung tissue such as Beas-2B, Calu-3, and A549, as well as novel human airway epithelial tissue, to elucidate important details about its life cycle in this model and identify potential factors that may affect HMPV infection in vivo. We hypothesize heparan sulfate proteoglycans, specifically the syndecan moieties, serve as an attachment factor for HMPV, which is required for internalization, and membrane fusion occurs in intracellular compartments. We expect heparan sulfate is also critical for viral infectivity in human airway epithelium. We also anticipate tissus derived from patients with chronic respiratory disease will show an increased permissibility to HMPV infection.

 描述（由适用提供）：人类元瘤病毒（HMPV）是最近发现的Paramyxoviridae家族中的呼吸道病原体，它感染了几乎100％的世界人群。这种包裹的病毒会导致婴儿，年龄较大和免疫功能低下的患者的严重病毒呼吸道疾病。实际上，HMPV估计是继密切相关的呼吸综合病毒之后的小儿下呼吸道疾病的第二大最常见原因。此外，65岁以上的成年人HMPV引起的呼吸道疾病的住院率接近影响力。尽管具有临床意义，但目前尚无抗病毒治疗或疫苗。自从发现其发现以来，已经确定了几个关键因素以在HMPV进入中发挥作用，但是结合，融合和进入的许多机制仍未解决。以前，我们的小组已经表明，硫酸乙酰肝素是HMPV结合和感染所需的部分。但是，尚不清楚乙par硫酸盐蛋白聚糖（HSPG）是否充当接收器，或者介导质膜上的其他必要的细胞因子。此外，尚未针对HMPV确定进入途径。为了阐明HSPG的作用，我们将利用实验室菌株和临床分离株的当前可用的HMPV菌株的集合来确定乙par硫酸盐阻断分子的影响以及敲低研究对特定HSPG的影响；为了识别进入途径，我们将利用带有荧光标记信封的病毒颗粒来跟踪膜膜是否发生在质膜或细胞内室内。最后，我们建议在传统细胞培养的背景下研究HMPV的进入，利用从人类肺组织（例如Beas-2b，Calu-3和A549）得出的物理相关细胞系，以及新型的人类气道上皮组织，阐明这种模型中有关其生命周期的重要细节，并识别可能影响其潜在因素的重要细节。 体内HMPV感染。我们假设硫酸乙酰肝素蛋白聚糖，特别是syndecan部分，是HMPV的附着因子，这是内在化所必需的，并且在细胞内室内发生膜融合。我们预计硫酸乙酰肝素对于人类气道上皮的病毒感染也至关重要。我们还期望从患有慢性呼吸系统疾病的患者中得出的组织将显示出增加HMPV感染的许可。