Neutrophil Extracellular Traps in Cystic Fibrosis

囊性纤维化(CF)仍然是一种不治之症，影响着全球7万人。缺乏新的CF疗法是 由于对疾病发病机制认识不深。肺部并发症是大多数CF的原因 死亡率。Cf呼吸道的特点是慢性细菌感染和强烈的白细胞渗透 叫做中性粒细胞。中性粒细胞释放其颗粒货物和DNA，从而导致肺损伤。 尽管中性粒细胞衍生的炎症介质的释放在CF中具有很高的临床相关性，但其 机制尚不清楚。该项目的长期目标是确定中性粒细胞 出于预防和治疗的目的，在CF中进行处理。此特定应用程序的目标是 探讨中性粒细胞胞外TRAP(Net)在CF中释放的机制及临床意义。这个 中心假说是铜绿假单胞菌鞭毛运动性增强，主要触发网状结构的形成 由呼吸道炎症环境决定，并与肺部疾病的临床措施有关。这 假设是基于申请人实验室提供的强有力的初步数据而提出的。篮网 由装饰着组蛋白和颗粒成分的DNA网络组成。建议的理由是 研究表明，一旦假单胞菌触发的网状形成机制和临床重要性将被 明确的，干扰它将提供一种新的方法来开发创新的新的CF疗法。这一假说将 通过追求以下具体目标进行测试：1)建立在CF气道中使用NETS的临床相关性 疾病，2)剖析铜绿假单胞菌触发净释放的机制，3)确定 Cf气道炎性环境对净形成的影响。根据我们的初步数据显示临床 在第一个目标中，将进行中性粒细胞标志物和中性粒细胞之间的相关性研究。 使用临床标本测量慢性肺病的严重程度。这些数据将揭示Nets是否可以 预测CF肺功能下降或与CF肺恶化有关。在第二个目标中，我们将 确定铜绿假单胞菌刺激网络形成的机制。在第三个目标中，我们将确定 炎症分子在CF中如何影响网络形成的机制。本研究具有创新性。 由于它使用了申请实验室开发的新工具来量化Net，它检测到了几个 CF临床标本中新的网络相关标记物，提示CF具有显著的自身免疫性 成分，它有可能识别新的候选CF生物标记物。拟议的研究是 意义重大，因为它关注的是一个临床上相关的，尚未解决的问题，通过使用原代人类细胞 和CF临床标本。总而言之，我们的提案将提供必要的数据，以对 我们将致力于研究CF呼吸道炎症领域，并能够设计出更好的CF治疗方法。

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.