Proteomics-based approach to ARDS

描述(由申请人提供)： 成人呼吸窘迫综合征(ARDS)是一种快速发展的严重呼吸衰竭的破坏性过程。ARDS的特点是急性炎症过程，上皮和内皮屏障受损。我们目前对肺损伤的基本发病机制的了解还不完全。我们预测高危患者发生肺损伤的能力以及一旦发生肺损伤的严重程度的能力很差。在血清或支气管肺泡灌洗液(BALF)中识别细胞和/或生化标志物的努力表明，单一的生化测定并不能一致地预测ARDS的发病或严重程度。这种方法无法识别未被怀疑的介体或新蛋白质。这项建议的目标是通过分析作为西雅图急性肺损伤ARDS SCOR计划的一部分获得的BALF，获得ARDS期间空中环境的全球蛋白质图谱。蛋白质组学中的新方法，如同位素编码的亲和标签和自动验证过程，将被用于分析BALF。这些技术使人们能够对复杂的生物样品进行定量和定性的蛋白质分析，并且可以用高通量设备进行。获得的蛋白质图谱将被用于制定分类方案，以帮助预测结果并帮助做出治疗决策。此外，参与ARDS发展的新的或未被怀疑的介体和调节通路可能被发现，并可能验证或挑战ARDS的机制。 最后，任何生成的模型和假说都将在急性肺损伤计划继续期间在ARDS患者中进行前瞻性测试。本研究的目的是：1.检测第3天ARDS患者和正常人BALF的蛋白质谱，以确定ARDS BALF中存在的可溶性蛋白质谱，并潜在地识别新的蛋白质。2.对同一患者的系列样本进行蛋白质谱分析，以获得ARDS发生发展过程中蛋白质表达的时程变化。3.比较进展为ARDS的高危人群和无进展的高危人群的蛋白质谱。4.比较ARDS幸存者和非ARDS幸存者的蛋白质图谱。这些结果将被用来开发肺损伤结果的预测模型。

DESCRIPTION (provided by applicant): Adult Respiratory Distress Syndrome (ARDS) is a devastating process of rapidly progressive and severe respiratory failure. ARDS is characterized by an acute inflammatory process with damage to the epithelial and endothelial barrier. Our current understanding of the basic pathogenic mechanisms involved in lung injury is incomplete. Our ability to predict the onset of lung injury in patients at risk and the severity of lung injury once it develops is poor. Efforts to identify cellular and/or biochemical markers for outcomes in serum or bronchoalveolar lavage fluid (BALF) has shown that single biochemical measurements are not consistent predictors of either the onset or the severity of ARDS. This approach fails to identify unsuspected mediators or new proteins. The goal of this proposal is to obtain global protein profiles of the airspace environment during ARDS by analyzing BALFobtained as part of the Seattle ARDS SCOR Program in acute lung injury. New methodologies in proteomics, such as isotope-coded affinity tag, and automated validation process, will be used to analyze BALF. These techniques enable one to obtain quantitative and qualitative protein analysis of complex biological specimens, and can be performed with a high throughput facility. The obtained protein profiles will be used to develop classification schemes that will assist in predicting outcome and assist in therapeutic decisions. Additionally, novel or unsuspected mediators and regulatory pathways involved in the development of ARDS may be discovered and may verify or challenge mechanisms of ARDS. Finally, any generated models and hypothesis will tested prospectively in ARDS patients during the continuation of the acute lung injury program. The goals of this study are to: 1. Examine the protein profile of BALF from day 3 ARDS patients, compared to normals in order to identify the spectrum of soluble proteins present in ARDS BALF and potentially identify new proteins. 2. Analyze the protein profiles from serial samples from the same patients in order to obtain a time course of protein expression during the development of ARDS. 3. Compare protein profiles from at risk that progressed to ARDS versus at risk with no progression. 4. Compare protein profiles from ARDS survivors vs. ARDS non-survivors. These results will be used to develop prediction models for the outcome of lung injury.