Purines as Biomarkers of Respiratory Disease

像囊性纤维化(CF)这样的呼吸系统疾病的特征是一系列的病理生理 从粘液分泌物脱水开始到粘液纤毛清除受损的变化， 慢性呼吸道炎症，以及随后的肺损伤。这些变化的有效生物标志物是 迫切需要监测疾病进展，识别恶化情况，并评估现有 和新奇的疗法。此职业发展申请的目标是为Charles Esther博士提供 识别和评估呼气冷凝液(EBC)中候选生物标志物所需的技能 即使在幼儿身上也可以简单、非侵入性地采集呼吸道样本。埃斯特博士计划 克服分析低浓度、可变稀释度EBC的挑战 液质联用(LC-MS/MS)。这项技术代表了一种高度 灵敏灵活的方法，允许同时测量多个小分子 生物标志物以及尿素作为稀释标志物。在这项研究提案中，Esther博士将使用MASS 光谱学鉴定和检测CF两个关键方面的简单和非侵入性EBC生物标记物 呼吸道病理生理学：中性粒细胞呼吸道炎症和呼吸道粘液分泌物脱水。 根据已建立的生物标志物和代谢组学的靶向分析确定潜在的生物标志物 中性粒细胞炎症和粘液脱水体外模型的研究。测量协议将 将使用LC-MS/MS开发，并在一组儿童和成人中评估有前景的生物标志物 确定它们的可靠性及其与公认的疾病病理生理学标志物的关系，并确定 潜在的混杂因素。成功完成本提案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工具，但将 还为Esther博士提供必要的技能和经验，以发展身份识别和 呼吸系统疾病的非侵入性生物标志物的检测。 相关性(请参阅说明)： 囊性纤维化的特征是肺内与疾病相关的变化，Esther博士建议 使用尖端技术开发简单且非侵入性的方法来检测这些变化 质谱仪和呼出的冷凝液收集。这些方法的发展将提供 囊性纤维化和其他肺部疾病的临床管理和研究的宝贵工具。

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.