Diagnostic and Prognostic Biomarkers in Pneumonia

肺炎的诊断和预后生物标志物

• 批准号: 7215800
• 负责人: ANTHONY F. SUFFREDINI
• 金额: --
• 依托单位: CLINICAL CENTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7215800
• 关键词: biomarker; bronchoscopy; clinical chemistry; diagnostic respiratory lavage; dogs; human subject; laboratory rabbit; mass spectrometry; matrix assisted laser desorption ionization; microarray technology; molecular biology information system; patient oriented research; pneumonia; prognosis; protein quantitation /detection; proteomics; two dimensional gel electrophoresis; biomarker; bronchoscopy; clinical chemistry; diagnostic respiratory lavage; dogs; human subject; laboratory rabbit; mass spectrometry; matrix assisted laser desorption ionization; microarray technology; molecular biology information system; patient oriented research; pneumonia; prognosis; protein quantitation /detection; proteomics; two dimensional gel electrophoresis

The objective of this study (04-CC-0119)is to analyze bronchoalveolar lavage and serum from patients with lung infiltrates in order to discover new biomarkers and protein expression patterns that are associated with specific types of pulmonary disease. Bronchoalveolar lavage is a standard method to obtain lower airway samples to evaluate pulmonary infiltrates in order to diagnose infection, malignancy or non-infectious inflammation. After collecting the lavage, the clinical microbiology laboratory concentrates the formed elements (i.e. pathogens and cells) for stains and culture and discards the bronchoalveolar lavage supernatant. The supernatant however is a rich source of proteins and other molecules. We hypothesize that bronchoalveolar lavage will be an important source of biomarkers that reflect host-pathogen interactions. The analysis of protein mass profiles and biomarker identification in bronchoalveolar lavage and serum may help develop new diagnostic methods and extend our understanding of mechanisms of lung inflammation due to infectious causes. The study population will include all patients undergoing bronchoscopy for clinical indications at the Clinical Center and affiliated study hospitals. This will facilitate the acquisition of BAL samples that reflect a spectrum of community-acquired and opportunistic pathogens associated with pulmonary disease. In addition analysis of a range of non-infectious pulmonary processes (e.g. acute lung injury, acute respiratory distress syndrome and engraftment syndrome) is important to develop measures of sensitivity and specificity. We currently use two different mass spectrometry platforms for analysis of biologic specimens including BAL fluid or serum/plasma. The Ciphergen Protein Chip Arrays (Ciphergen Biosystems, Inc., Palo Alto, CA, USA) selectively fractionates samples based on binding affinity to specialized surfaced including hydrophobic, cation, anion and immobilized metal affinity capture. This technique of surface chemistry optimization is termed Surface Enhanced Laser Desorption-Ionization (SELDI) and when combined with mass spectrometry, termed SELDI-TOF. We will complement our SELDI protein expression profiles with two-dimensional gel electrophores and a higher resolution mass spectrometry system (Bruker Daltonics Ultraflex MS/MS). By developing a large database of BAL fluid linked to specific microbiologic diagnoses, we plan to define protein expression signature response profiles that distinguish specific etiologies of lung infection and inflammation. These signature profiles will be based on mass spectrometry, two-dimensional gel electrophoresis and suspension array technologies. Because of the variability associated with individual host responses to infection due to differences in host immunity, sampling time effects, and external factors such as antibiotic or anti-inflammatory therapies, a large database will be required. The profiles of culture-negative BAL fluid will be of similar interest to assist in defining non-infectious etiologies of lung inflammation. A secondary objective is to perform proteomic analysis on serum collected from patients at the time of bronchoscopy. The goal is to link serum proteomic profiles to BAL proteomic profiles to determine whether a less invasive technique can predict infiltrate etiology with comparable sensitivity and specificity to BAL profiles. To complement the patient studies we will investigate protein biomarkers in blood and lavage from animal models of pneumonia. We are currently studying a rabbit model of invasive pulmonary aspergillosis and a canine model of staphylococcal pneumonia. Exploring these model systems will facilitate our identification of candidate biomarkers across species. A total of 240 patients have been enrolled in this study to date. Approximately one half of the participants have a specific microbiologic diagnosis as a cause of their pulmonary infiltrates. The bronchoalveolar lavage and blood samples are currently being analyzed.

这项研究的目的（04-CC-0119）是分析肺浸润患者的支气管肺泡灌洗和血清，以发现与特定类型的肺疾病相关的新生物标志物和蛋白质表达模式。支气管肺泡灌洗是一种标准方法，可以获取较低的气道样品以评估肺部浸润，以诊断感染，恶性肿瘤或非感染性炎症。收集灌洗后，临床微生物学实验室将形成的元素（即病原体和细胞）集中在污渍和培养物中，并丢弃支气管肺泡灌洗上清液。但是，上清液是蛋白质和其他分子的丰富来源。我们假设支气管肺泡灌洗将是反映宿主 - 病原体相互作用的重要来源。支气管肺泡灌洗和血清中蛋白质质量谱和生物标志物鉴定的分析可能有助于开发新的诊断方法，并扩展我们对由于传染性原因引起的肺部炎症机制的理解。 研究人群将包括所有在临床中心和附属研究医院进行支气管镜检查的患者。这将有助于获取BAL样品，这些BAL样品反映了与肺部疾病相关的一系列社区获得的和机会性病原体。另外，分析了一系列非感染性肺部过程（例如急性肺损伤，急性呼吸窘迫综合征和植入综合征），对于制定敏感性和特异性的度量很重要。 目前，我们使用两个不同的质谱平台来分析包括BAL流体或血清/血浆在内的生物样品。 Ciphergen蛋白芯片阵列（Ciphergen Biosystems，Inc.，Palo Alto，CA，美国）选择性分数基于与专业表面的结合亲和力，包括疏水，阳离子，阴离子，阴离子和固定的金属亲和力捕获。这种表面化学优化的技术称为表面增强的激光解吸收离子（SELDI），当与质谱法结合使用时，称为SELDI-TOF。我们将与二维凝胶电泳和更高分辨率的质谱系统（Bruker Daltonics Ultraflex MS/MS）相互补充Seldi蛋白表达谱。 通过开发与特定微生物学诊断相关的大型BAL流体数据库，我们计划定义蛋白质表达的特征反应曲线，以区分肺部感染和炎症的特定病因。这些特征曲线将基于质谱，二维凝胶电泳和悬架阵列技术。由于与宿主免疫，抽样时间效应的差异以及抗生素或抗炎疗法等外部因素有关的单个宿主对感染的反应的可变性，因此需要大量数据库。培养阴性BAL液的特征将具有相似的兴趣，以帮助定义肺部炎症的非感染病因。 次要目标是对支气管镜检查时从患者收集的血清进行蛋白质组学分析。目的是将血清蛋白质组学谱与BAL蛋白质组织谱链接起来，以确定侵入性较低的技术是否可以预测浸润性病因，具有可比的敏感性和对BAL谱的特异性。 为了补充患者研究，我们将研究肺炎动物模型的血液和灌洗中的蛋白质生物标志物。我们目前正在研究侵入性肺曲霉病和葡萄球菌肺炎犬模型的兔模型。探索这些模型系统将有助于我们识别跨物种的候选生物标志物。 迄今为止，总共已有240名患者参加了这项研究。大约一半的参与者具有特定的微生物诊断，是其肺部浸润的原因。目前正在分析支气管肺泡灌洗和血液样本。