Neutrophil Extracellular Traps in Cystic Fibrosis

囊性纤维化（CF）仍然是一种无法治愈的疾病，影响全球70，000人。缺乏新的CF疗法是 由于对疾病的发病机理认识不足。肺部并发症是大多数CF的原因 mortality. CF气道的特征是慢性细菌感染和白细胞的强烈浸润 叫做中性粒细胞。中性粒细胞释放其颗粒货物和DNA以引起肺损伤。 尽管嗜中性粒细胞衍生的炎症介质的释放在CF中具有高度的临床相关性，但其 机制不明。这个项目的长期目标是确定中性粒细胞如何被 用于预防和治疗目的。本申请的目的是 确定CF中中性粒细胞胞外陷阱（NET）释放的机制和临床相关性。的 中心假设是CF中的NET形成主要由铜绿假单胞菌鞭毛运动触发，增强 通过气道炎症环境，并与肺部疾病的临床措施。这 假设是基于申请人实验室中产生的强有力的初步数据而制定的。网 是由一个由组蛋白和颗粒成分装饰的DNA网组成的。建议的理由 研究表明，一旦假单胞菌触发NET形成的机制和临床重要性将被 很明显，干扰它将为开发创新的CF疗法提供一种新的方法。这一假设将 通过追求以下特定目标进行测试：1）确定CF气道中NETs的临床相关性 疾病，2）剖析P. aerodosa-triggered NET释放的机制，以及3）确定 CF对NET形成的气道炎症环境的影响。根据我们的初步数据显示， 在CF中NETs的相关性方面，第一个目标是，将在中性粒细胞标志物和 使用CF临床样品测量CF肺病严重程度。这些数据将揭示NET是否可以 预测CF肺功能下降或与CF肺恶化相关。第二个目标，我们将 确定铜绿假单胞菌刺激NET形成的机制。在第三个目标中，我们将确定 CF中存在的炎症分子如何影响NET形成的机制。这项研究具有创新性 因为它使用了申请实验室开发的新工具来定量NET，所以它检测到了几种 新的NET相关标志物在CF临床样本，它表明CF有显着的自身免疫性 这是一种新的CF生物标志物组分，并且它具有鉴定新的CF生物标志物候选物的潜力。拟议的研究是 重要的是，它通过使用原代人类细胞关注临床相关的未解决的CF问题 和CF临床样品。总之，我们的提案将提供必要的数据，以提供对 CF气道炎症领域，并能够设计更好的CF疗法。

Cystic fibrosis (CF) is still an incurable disease affecting 70,000 people worldwide. Lack of new CF therapies is due to poor understanding of disease pathogenesis. Lung complications are responsible for majority of CF mortality. CF airways are characterized by chronic bacterial infections and robust infiltration of leukocytes called neutrophil granulocytes. Neutrophils release their granule cargo and DNA to cause lung damage. Although release of neutrophil-derived inflammatory mediators is of high clinical relevance in CF, its mechanism is unknown. The long-term goal of this project is to determine how neutrophils could be manipulated in CF for preventive and therapeutic purposes. The objective in this particular application is to determine the mechanism and clinical relevance of neutrophil extracellular trap (NET) release in CF. The central hypothesis is that NET formation in CF is mainly triggered by P. aeruginosa flagellar motility, enhanced by the airway inflammatory environment and is associated with clinical measures of lung disease. This hypothesis has been formulated based on strong preliminary data produced in the applicant's laboratory. NETs are composed of a DNA web decorated with histones and granule components. The rationale for the proposed research is that, once the mechanism and clinical importance of Pseudomonas-triggered NET formation will be clear, interfering with it will offer a novel approach to develop innovative new CF therapies. This hypothesis will be tested by pursuing the following specific aims: 1) Establish the clinical relevance of NETs in CF airway disease, 2) dissect the mechanism of P. aeruginosa-triggered NET release, and 3) determine the effect of the CF airway inflammatory environment on NET formation. Based on our preliminary data showing clinical relevance of NETs in CF, in the first aim, correlation studies will be performed between neutrophil markers and measures of CF lung disease severity using CF clinical samples. These data will reveal whether NETs can predict CF lung function decline or are linked to CF pulmonary exacerbations. In the second aim, we will identify the mechanism of P. aeruginosa-stimulated NET formation. In the third aim, we will determine the mechanism how inflammatory molecules present in CF affect NET formation. This research is innovative because it uses novel tools developed by the applicant laboratories to quantitate NETs, it detected several novel NET-related markers in CF clinical samples, it suggests that CF has a significant autoimmune component, and it has the potential to identify new CF biomarker candidates. The proposed research is significant because it focuses on a clinically relevant, unsolved question of CF by using primary human cells and CF clinical samples. In summary, our proposal will deliver essential data to provide a major impact on the field of CF airway inflammation and to be able to design better CF therapies.