Center for Lung Biology, University of Washington, Seattle, WA 98109, USA.
J Allergy Clin Immunol. 2011 Dec;128(6):1176-1184.e6. doi: 10.1016/j.jaci.2011.07.053. Epub 2011 Sep 8.
Asthma is a heterogeneous disease characterized by abnormal airway pathophysiology and susceptibility to different stimuli, as exemplified by a subset of patients with exercise-induced bronchoconstriction. Induced sputum provides a noninvasive method to sample airway biofluids that are enriched in proteins.
We hypothesized that novel mechanisms in the pathogenesis of asthma might be revealed by studying the patterns of protein expression in induced sputum.
We used shotgun proteomics to analyze induced sputum from 5 healthy subjects and 10 asthmatic patients, including 5 with exercise-induced bronchoconstriction. Differential protein expression among asthmatic patients, asthma subphenotypes, and control subjects was determined by using spectral counting and computational methods.
Using Gene Ontology analysis, we defined the functional landscape of the induced sputum proteome and applied network analysis to construct a protein interaction map for this airway compartment. Shotgun proteomics analysis identified a number of proteins the differential enrichment or depletion of which robustly distinguished asthmatic patients from healthy control subjects and captured the effects of exercise on induced sputum proteome. Functional and network analysis identified key processes, including proteolytic activity, that are known contributors to airway remodeling. Importantly, this approach highlighted previously unrecognized roles for differentially expressed proteins in pathways implicated in asthma, such as modulation of phospholipase A(2) by secretoglobin, a putative role for S100A8/9 in human asthma, and selective upregulation of complement component 3a in response to exercise in asthmatic patients.
Computationally intensive analysis of induced sputum proteome is a powerful approach to understanding the pathophysiology of asthma and a promising methodology to investigating other diseases of the airways.
哮喘是一种异质性疾病，其特征为气道病理生理学异常和对不同刺激的易感性，例如一部分运动诱发性支气管收缩患者。诱导痰提供了一种非侵入性的方法来采集富含蛋白质的气道生物液。
我们假设通过研究诱导痰中蛋白质表达模式，可以揭示哮喘发病机制中的新机制。
我们使用鸟枪法蛋白质组学分析了 5 名健康受试者和 10 名哮喘患者（包括 5 名运动诱发性支气管收缩患者）的诱导痰。通过光谱计数和计算方法确定哮喘患者、哮喘亚表型和对照受试者之间的差异蛋白表达。
使用基因本体论分析，我们定义了诱导痰蛋白质组的功能景观，并应用网络分析构建了该气道隔室的蛋白质相互作用图。鸟枪法蛋白质组学分析鉴定了许多蛋白，其差异丰度或耗竭可稳健地区分哮喘患者与健康对照受试者，并捕获了运动对诱导痰蛋白质组的影响。功能和网络分析确定了关键过程，包括蛋白水解活性，这些过程已知是气道重塑的贡献因素。重要的是，这种方法突出了差异表达蛋白在哮喘相关途径中的先前未被认识的作用，例如分泌球蛋白对磷脂酶 A2 的调节、S100A8/9 在人类哮喘中的潜在作用以及补体成分 3a 在哮喘患者运动时的选择性上调。
对诱导痰蛋白质组进行计算密集型分析是理解哮喘病理生理学的一种有力方法，也是研究气道其他疾病的一种有前途的方法。
