Purines as Biomarkers of Respiratory Disease

描述(申请人提供)：囊性纤维化(CF)等呼吸道疾病的特征是一系列的病理生理变化，从粘液分泌脱水开始，发展到粘液纤毛清除受损，慢性呼吸道炎症，以及随后的肺损伤。迫切需要这些变化的有效生物标志物来监测疾病进展，识别病情恶化，并评估现有和新疗法的疗效。这一职业发展应用程序的目标是为Charles Esther博士提供所需的技能，以识别和评估呼气冷凝液(EBC)中的候选生物标记物，EBC是一种即使在幼儿身上也可以简单且非侵入性地采集的呼吸道样本。Esther博士计划通过应用LC-MS/MS(LC-MS/MS)来克服分析低浓度、可变稀释度EBC的挑战。这项技术代表了一种高度灵敏和灵活的方法，允许同时测量多个小分子生物标记物以及尿素作为稀释标记物。在这项研究方案中，Esther博士将使用质谱仪识别和测试CF气道病理生理的两个关键方面的简单且非侵入性的EBC生物标志物：中性粒细胞气道炎症和呼吸道粘液分泌脱水。基于对已建立的生物标志物的靶向分析和对中性粒细胞炎症和粘液脱水的体外模型的代谢组学研究，确定了潜在的生物标志物。将使用LC-MS/MS开发测量方案，并在一组儿童和成人中评估有希望的生物标记物，以确定其可靠性及其与公认的疾病病理生理学标记物的关系，并确定潜在的混杂因素。成功完成这项建议中概述的研究计划不仅将为未来关于CF的病理生理学和治疗选择的研究提供重要的工具，而且还将为Esther博士提供必要的技能和经验，以发展呼吸系统疾病的非侵入性生物标志物的识别和测试事业。相关性(参见说明)：囊性纤维化的特征是肺内疾病相关的变化，Esther博士建议开发简单且非侵入性的方法，使用质谱仪和呼气冷凝液收集等尖端技术来检测这些变化。这些方法的发展将为囊性纤维化和其他肺部疾病的临床管理和研究提供宝贵的工具。(摘要结束)

DESCRIPTION (provided by applicant): Respiratory diseases such as cystic fibrosis (CF) are characterized by a cascade of pathophysiological changes starting with dehydration of mucus secretions and proceeding to impaired mucociliary clearance, chronic airway inflammation, and subsequent lung injury. Effective biomarkers of these changes are urgently needed to monitor disease progression, identify exacerbations, and evaluate the efficacy of existing and novel therapies. The goal of this career development application is to provide Dr. Charles Esther with the skills needed to identify and assess candidate biomarkers in exhaled breath condensate (EBC), an airway sample that can be collected simply and non-invasively even in young children. Dr. Esther plans to overcome the challenges of analyzing the low concentration, variable dilution EBC through application of liquid-chromatography-tandem mass spectrometry (LC-MS/MS). This technology represents a highly sensitive and flexible methodology that permits simultaneous measurement of multiple small molecule biomarkers as well as urea as a dilution marker. In this research proposal, Dr. Esther will use mass spectrometry to identify and test simple and non-invasive EBC biomarkers of two critical aspects of CF airways pathophysiology: neutrophilic airway inflammation and dehydration of airway mucus secretions. Potential biomarkers identified based on both targeted analyses of established biomarkers and metabolomic studies of in vitro models of neutrophilic inflammation and mucus dehydration. Measurement protocols will be developed using LC-MS/MS, and promising biomarkers assessed in a cohort of children and adults with to determine their reliability and relationship to accepted markers of disease pathophysiology and to identify potential confounders. Successful completion of the research plan outlined in this proposal will not only generate important tools for future research on the pathophysiology and therapeutic options for CF, but will also provide Dr. Esther with the skills and experiences necessary to develop a career in identification and testing of non-invasive biomarkers of respiratory disease. RELEVANCE (See instructions): Cystic fibrosis is characterized by disease related changes within the lung, and Dr. Esther proposes to develop simple and non-invasive methods to detect these changes using the cutting edge technologies of mass spectrometry and exhaled breath condensate collection. Development of these methods will provide invaluable tools for clinical management and research in cystic fibrosis and other lung diseases. (End of Abstract)