Contribution of neutrophils to early airway disease in cystic fibrosis children

中性粒细胞对囊性纤维化儿童早期气道疾病的影响

• 批准号: 9143792
• 负责人: Rabindra Marie-Jean Tirouvanziam
• 金额: $ 33.37万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-14 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9143792
• 关键词: Affect; Air; Ancillary Study; Apical; Bacteria; Biological Assay; Biological Markers; Bronchiectasis; Bronchoalveolar Lavage Fluid; CASP1 gene; Caucasians; Cell Death; Cells; Child; Chronic; Clinical; Cystic Fibrosis; Cytoplasmic Granules; Data; Development; Disease; Drug Targeting; Early Intervention; Enzymes; Flow Cytometry; Functional disorder; Goals; Health; Human; Image Cytometry; In Vitro; Infant; Infection; Inflammation; Intervention; Leukocyte Elastase; Lipids; Liquid substance; Logistic Regressions; Mass Spectrum Analysis; Measures; Metabolic; Metabolic Activation; Methods; Modeling; Morbidity - disease rate; Mucous body substance; Netherlands; Neutrophil Infiltration; Obstruction; Organ; Parents; Pathway interactions; Patients; Peroxidases; Phagocytosis; Phagosomes; Phase; Play; Preclinical Drug Evaluation; Process; Recruitment Activity; Regression Analysis; Research; Research Personnel; Respiratory physiology; Role; Sampling; Testing; Time; X-Ray Computed Tomography; abstracting; airway inflammation; base; burden of illness; children with cystic fibrosis; cohort; cystic fibrosis airway; cystic fibrosis patients; early cystic fibrosis; extracellular; in vitro Model; in vivo; innovation; insight; mortality; multidisciplinary; neutrophil; new therapeutic target; pre-clinical; prevent; prospective; therapy development; tool

 DESCRIPTION (provided by applicant): Hallmarks of cystic fibrosis (CF) airway disease include bronchiectasis, inflammation by polymorphonuclear neutrophils (PMNs) from blood, obstruction by mucus, and infection by bacteria. In landmark studies, the AREST CF consortium showed that first signs of bronchiectasis in CF children occur in small airways and coincide with PMN recruitment and high activity of neutrophil elastase (NE) in the airway fluid. Recently, we showed that a large fraction of airway PMNs in chronic CF disease release NE while viable, as part of a reprogramming process that includes metabolic activation, caspase-1 activity and hyperexocytosis. Here, we show that similar hyperexocytosis occurs in airway PMNs from CF children. In addition, we introduce a new model of human small airway in which naive blood PMNs exposed to apical CF airway fluid in vitro are induced to transmigrate and become hyperexocytic, recapitulating features of CF airway PMNs in vivo. Our long-term goal is to develop therapies aimed at correcting PMN dysfunction, ideally via targeted intervention in early CF disease. As a step toward this objective, we will conduct a mechanistic study of PMN function in early CF leveraging: (i) our renewed understanding of PMN dysfunction in CF; (ii) our new in vitro model, and (iii) the unique AREST-CF study about to be initiated in the Netherlands. This prospective cohort is modeled after the Australian AREST-CF study, including computed tomography (CT) and airway fluid analysis at 12, 36 and 60 months to closely follow the onset and progression of early CF disease. Our overall hypothesis is that early CF disease coincides with airway PMN reprogramming. Our aims are to: AIM 1- Determine the extent of PMN reprogramming in early CF disease, in vivo. Hypothesis 1A: Airways in CF children contain reprogrammed PMNs. Hypothesis 1B: Progression of airway disease measured clinically in CF children associates with higher burden of reprogrammed PMNs. AIM 2- Identify mechanisms of PMN reprogramming by CF airway fluid, in vitro. Hypothesis 2A: Airway fluid from CF children induces PMN reprogramming in our model. Hypothesis 2B: Modulation of functional pathways in reprogrammed airway PMNs can inhibit their dysfunction and associated disease burden. To leverage the extraordinary research opportunity afforded by the parent study, we assembled a group of CF researchers with multidisciplinary expertise. This proposed ancillary study is highly time-sensitive, since the parent study will start shortly, and high-content assays included here (e.g., flow cytometry, image cytometry, multiplexed qPCR, targeted mass spectrometry) require immediate processing of patient samples. We expect this study to yield better mechanistic understanding of early CF disease and strong proof- of-concept data on new targeted therapies to help prevent the vicious cycle of CF airway inflammation.

 描述（由申请人提供）：囊性纤维化（CF）气道疾病的特征包括支气管扩张、血液中多形核中性粒细胞（PMN）引起的炎症、粘液阻塞和细菌感染。在具有里程碑意义的研究中，AREST CF联盟显示CF儿童支气管扩张的最初体征发生在小气道中，与气道液中PMN募集和中性粒细胞弹性蛋白酶（NE）高活性一致。最近，我们发现，在慢性CF疾病中，大部分气道中性粒细胞在存活时释放NE，作为重编程过程的一部分，包括代谢活化、半胱天冬酶-1活性和胞饮过多。在这里，我们表明，类似的过度分泌发生在气道中性粒细胞从CF儿童。此外，我们引入了一种新的人类小气道模型，其中体外暴露于顶部CF气道液体的幼稚血液中性粒细胞被诱导迁移并成为体内CF气道中性粒细胞的过度氧化、重演特征。我们的长期目标是开发旨在纠正PMN功能障碍的治疗方法，理想情况下通过早期CF疾病的靶向干预。作为实现这一目标的一步，我们将进行一项机制研究的PMN功能在早期CF利用：（一）我们的PMN功能障碍CF的新认识;（二）我们的新的体外模型，（三）独特的AREST-CF研究即将在荷兰启动。该前瞻性队列以澳大利亚AREST-CF研究为模型，包括12、36和60个月时的计算机断层扫描（CT）和气道液体分析，以密切跟踪早期CF疾病的发作和进展。我们的总体假设是，早期CF疾病与气道PMN重编程一致。我们的目的是：目的1-确定在体内的早期CF疾病的PMN重编程的程度。假设1A：CF儿童的气道含有重编程的PMNs。假设1B：CF儿童临床测量的气道疾病进展与重编程的PMNs的较高负担相关。目的2-在体外通过CF气道液确定PMN重编程的机制。假设2A：在我们的模型中，CF儿童的气道液体诱导PMN重编程。假设2B：调节重编程气道中性粒细胞的功能通路可以抑制其功能障碍和相关的疾病负担。为了利用母研究提供的非凡研究机会，我们召集了一组具有多学科专业知识的CF研究人员。由于母研究将很快开始，并且此处包含的高含量测定（例如，流式细胞术、图像细胞术、多重qPCR、靶向质谱法）需要立即处理患者样品。我们希望这项研究能够更好地了解早期CF疾病的机制，并提供关于新靶向治疗的强有力的概念验证数据，以帮助预防CF气道炎症的恶性循环。