Altered hyaluronan deposition by bronchial epithelial cells contributes to inflammation during respiratory syncytial virus infection

支气管上皮细胞改变的透明质酸沉积导致呼吸道合胞病毒感染期间的炎症

• 批准号: 10418432
• 负责人: STEPHEN R REEVES
• 金额: $ 9.43万
• 依托单位: SEATTLE CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10418432
• 关键词: Acute; Adhesions; Adhesives; Alveolus; Attenuated; Binding; Biological Assay; Biological Models; Cell Culture Techniques; Cell Line; Cells; Child; Childhood; Coculture Techniques; Data; Deposition; Epithelial Cells; Extracellular Matrix; Fibroblasts; Future; Genes; Hospitalization; Human; Hyaluronan; Immune; Immune response; Infection; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Response; Knowledge; Laboratories; Lead; Leukocytes; Lower Respiratory Tract Infection; Lung; Mediating; Modeling; Modification; Molecular; Morbidity - disease rate; Mucous body substance; Neutrophil Activation; Obstruction; Oligonucleotides; Pathologic; Pharmacology; Play; Positioning Attribute; Production; Proteins; Pulmonary Inflammation; Reporting; Respiratory Disease; Respiratory Syncytial Virus Infections; Respiratory Therapy; Respiratory syncytial virus; Role; Shapes; Site; Small Interfering RNA; Source; Stromal Cells; Structure of parenchyma of lung; Submucosa; Supportive care; TNF gene; Techniques; Testing; Therapeutic Intervention; Up-Regulation; Vaccines; Viral Pathogenesis; Viral Respiratory Tract Infection; Virus Diseases; White Blood Cell Count procedure; Work; acute infection; airway inflammation; airway obstruction; bronchial epithelium; cell injury; experimental study; in vivo; inhibitor; knock-down; leukocyte activation; mortality; neutrophil; new therapeutic target; novel; novel therapeutic intervention; novel therapeutics; pathogenic virus; recruit; respiratory virus; response to injury; small hairpin RNA

PROJECT SUMMARY / ABSTRACT Lower respiratory tract infections commonly caused by respiratory syncytial virus (RSV) are a leading cause of hospitalizations and mortality worldwide in young children. Despite many years of study, the mainstay of treatment remains supportive care and currently no RSV specific treatment or vaccine is available. The cells that line the airways, otherwise known as bronchial epithelial cells (BECs), are the primary target for RSV infection. Infection of the BECs with RSV leads to subsequent BEC damage, obstruction of the lower airways with cellular debris and mucous, and the establishment of airway inflammation by recruitment of immune cells such as neutrophils into the peribronchial space. Recent studies from our laboratory have demonstrated that RSV infection of stromal cells such as lung fibroblasts leads to the establishment of an extracellular matrix (ECM) that is enriched with hyaluronan (HA) which promotes the accumulation and activation of leukocytes in ex vivo cell culture models. Furthermore, we found that the increased HA accumulation following RSV infection was more closely associated with the fibroblast cell layer and displayed greater modification with heavy chains (HC) which has been described in other studies to enhance the ECM’s ability to become sticky for inflammatory cells and promotes the inflammatory response in the lung. The formation of HC-HA is the result of the enzymatic activity of tumor necrosis factor stimulated gene 6 (TSG-6), which is not normally expressed in healthy lung tissue, but is induced in response to injury. In our previous work with lung fibroblasts, we have shown that TSG-6 is upregulated during RSV infection and blocking its induction with siRNA decreased the amount of HC-HA that was formed and decreased the accumulation of leukocytes. Despite being the primary target of RSV infection, no studies exist that have evaluated the effects of RSV infection on BEC production of HA or TSG-6 induced HC-HA formation despite the important role that HC-HA plays in promoting lung inflammation. Our laboratory both has expertise in the characterization of HA matrices and access to primary human BECs from well-characterized pediatric donors placing our group in a unique position to evaluate the contribution of BEC derived HC-HA to the inflammatory response following RSV infection. We hypothesize that RSV infection of BECs leads to increased production and accumulation of HA which will in turn promote the accumulation and activation of neutrophils in an ex vivo human cell culture model system. Additionally, we will test the hypothesis that RSV infection of BECs will induce the expression of TSG-6 thereby promoting the formation of HC-HA and further drive the accumulation and activation of neutrophils in our model system. We will pharmacologically block the formation of HC-HA enriched ECMs and also block the induction of TSG-6 during RSV infection in order to establish whether the increased accumulation of the neutrophils is HC-HA dependent. If true, characterizing the importance of HC-HA produced by BECs may lead to novel targets for therapeutic intervention during acute RSV infections and may be applicable to other respiratory viruses.

项目摘要 /摘要 通常由呼吸道合胞病毒（RSV）引起的下呼吸道感染是主要原因 全世界的住院和死亡率。尽管学习了多年，但 治疗仍然受支持的护理，目前尚无RSV特定治疗或疫苗可用。细胞 那条线，否则称为支气管上皮细胞（BEC）是RSV的主要目标 感染。带有RSV的BEC感染导致BEC损害，下呼吸道的目标 带有细胞碎片和粘液，以及通过募集免疫细胞来建立气道注射 例如中性粒细胞进入周围空间。我们实验室的最新研究表明 基质细胞（例如肺成纤维细胞）的RSV感染导致建立细胞外基质 （ECM）富含氢龙（HA），促进白细胞的积累和激活 离体细胞培养模型。此外，我们发现RSV感染后HA积累增加 与成纤维细胞层密切相关，并显示出更大的修饰。 （HC）在其他研究中描述了，以增强ECM的粘性能力 炎症细胞并促进肺中的炎症反应。 HC-HA的形成是结果 肿瘤坏死因子刺激基因6（TSG-6）的酶活性，通常不表达 在健康的肺组织中，但会因损伤而诱导。在我们以前使用肺成纤维细胞的工作中，我们有 表明在RSV感染期间更新TSG-6，并通过siRNA阻止其诱导量降低了 形成并改善白细胞积累的HC-HA量。尽管是主要 RSV感染的靶点，没有研究评估RSV感染对BEC生产的影响 HA或TSG-6诱导HC-HA的形成，尽管HC-HA在促进肺中起着重要作用 炎。我们的实验室都有在HA物品的表征和获得主要的特征方面的专业知识 来自特征良好的儿科捐赠者的人类BEC将我们的小组处于独特的位置，以评估 RSV感染后，BEC衍生的HC-HA对炎症反应的贡献。我们假设这一点 BEC的RSV感染导致HA的产量和积累增加，这反过来促进 在体内人类细胞培养模型系统中中性粒细胞的积累和激活。此外，我们会的 测试BEC的RSV感染将诱导TSG-6的表达的假设，从而促进 HC-HA的形成并进一步推动了模型系统中嗜中性粒细胞的积累和激活。我们 将实际阻止HC-HA富集的ECM的形成，并阻止TSG-6的诱导 在RSV感染期间，以确定中性粒细胞的积累增加是HC-HA 依赖。如果是真的，则表征BEC生产的HC-HA的重要性可能会导致新的目标 急性RSV感染期间的治疗干预措施，可能适用于其他呼吸道病毒。