RSV, Keratins and Distal Airway Infection

 描述（由申请方提供）：呼吸道合胞病毒（RSV）是婴儿毛细支气管炎的主要原因。RSV感染的人婴儿细支气管通常被脱落的RSV感染的上皮细胞堵塞，这些上皮细胞沿着嗜中性粒细胞占优势的炎性细胞浸润，导致远端气道急性阻塞。我们的长期目标是了解为什么RSV具有引起人类婴儿细支气管炎的倾向，并确定降低RSV远端气道疾病严重程度的治疗方法。我们最近已经鉴定了RSV非结构蛋白2（NS2）负责RSV感染的柱状上皮细胞的脱落，提供了上皮细胞脱落可能是RSV感染的特定结果的直接证据。使用仓鼠作为气道感染的体内模型，我们已经证明RSV NS2促进的细胞脱落到远端细支气管气道腔中导致脱落和病毒感染的上皮细胞的大量积累，这些上皮细胞堵塞和阻塞受感染的远端气道。由于仓鼠远端气道中RSV NS2表达的这些病理学结果与从患有RSV细支气管炎的人类婴儿获得的组织病理学惊人地相似，我们提出RSV NS2是RSV感染期间远端气道细胞病理学和疾病的重要病毒遗传决定因素。在这项初步和可行性研究中，我们专注于开发更好的模型，以进一步了解RSV NS2表达的后果，从而能够测试调节细胞脱落反应的新型疗法。我们还试图鉴定RSV NS2在远端气道中表达的新生物标志物，其可用于确定实验室和临床中新治疗方法的功效。改变对RSV NS2的细胞脱落反应的策略可能为降低RSV细支气管炎的发病率和严重程度提供新的治疗方法。我们进行了三个独立但相互关联的目的：具体目的1：RSV NS2表达的后果是否在幼仓鼠的远端气道中更持久和更严重？具体目标2。生成精密切割肺切片以研究RSV NS2表达在离体细支气管气道上皮中的影响。具体目标3。RSV NS2对上皮角蛋白的修饰是否为远端气道感染提供了新的生物标志物？通过执行这些目标，我们期望产生更好的模型来理解RSV NS2表达对远端气道病理学的影响，并鉴定用于评估RSV感染期间远端气道上皮受累的新型生物标志物。这些目标的成功完成将作为未来研究的基础，以确定减少RSV NS2表达后果影响的策略。

 DESCRIPTION (provided by applicant): Respiratory Syncytial Virus (RSV) is the major cause of bronchiolitis in infants. Human infant bronchioles infected by RSV are commonly noted to be occluded by sloughed, RSV-infected epithelial cells which along with neutrophil-dominant inflammatory cell infiltrates contribute to acute obstruction of the distal airways. Our long term goals are to understand why RSV has such a propensity for causing bronchiolitis in human infants and to identify therapeutic approaches for reducing the severity of RSV distal airway disease. We have recently identified the RSV Non-Structural protein 2 (NS2) as being responsible for the shedding of RSV-infected columnar epithelial cells providing direct evidence that epithelial cell shedding may be a specific consequence of RSV infection. Using hamsters as an in vivo model of airway infection we have demonstrated RSV NS2-promoted cell shedding into the distal bronchiolar airway lumen results in robust accumulation of shed and virus-infected epithelial cells which clog and obstruct the infected distal airways. As these pathological consequences of RSV NS2 expression in hamster distal airways are strikingly similar to the histopathology obtained from human infants with RSV bronchiolitis we propose RSV NS2 is an important viral genetic determinant for distal airway cytopathology and disease during RSV infection. In this Pilot and Feasibility study, we focus on the development of better models for further understanding the consequences of RSV NS2 expression to enable testing of novel therapies to modulate the cell shedding response. We also attempt to identify novel biomarkers of RSV NS2 expression in the distal airways useful for determining efficacy of novel therapeutic approaches in the lab and in the clinic. Strategies to modify the cell shedding response to RSV NS2 may provide novel therapeutic approaches for reducing the incidence and severity of RSV bronchiolitis. We perform three independent but inter-related Aims: Specific Aim 1: Are the consequences of RSV NS2 expression more prolonged and severe in the distal airways of infant hamsters? Specific Aim 2. Generation of Precision Cut Lung Slices to investigate the impact of RSV NS2 expression in the bronchiolar airway epithelium ex vivo. Specific Aim 3. Does RSV NS2 modification of epithelial keratins provide novel biomarkers for distal airway infection? By performing these Aims we expect to generate better models for understanding the impact of RSV NS2 expression on distal airway pathology and identify novel biomarkers for assessing distal airway epithelium involvement during RSV infection. Successful completion of these Aims will serve as the foundation of future studies to identify strategies to reduce the impact of the consequences of RSV NS2 expression.