Proteomic Analysis of Primary Human Airway Epithelial Cells Exposed to the Respiratory Toxicant Diacetyl.
Proteomic Analysis of Primary Human Airway Epithelial Cells Exposed to the Respiratory Toxicant Diacetyl.
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DOI：
10.1021/acs.jproteome.6b00672
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发表时间：
2017-02-03
影响因子：
4.4
通讯作者：
Palmer SM
中科院分区：
文献类型：
--
作者：
Foster MW;Gwinn WM;Kelly FL;Brass DM;Valente AM;Moseley MA;Thompson JW;Morgan DL;Palmer SM
职业性暴露于二酮调味剂双乙酰与闭塞性毛细支气管炎有关，这是一种罕见的气道纤维化疾病。啮齿类动物的模型研究表明，气道上皮是双乙酰毒性的主要部位，但双乙酰暴露对人类气道上皮的影响尚不清楚。在这里，我们采用定量LC-MS/MS-based蛋白质组学研究了重复暴露于双乙酰蒸汽对人原代气管支气管上皮细胞三维器官型培养的影响。使用无标记方法，我们在四个独立供体的细胞裂解物中量化了大约3400种蛋白质和5700种磷酸肽。蛋白和磷酸肽表达的改变提示了二乙酰暴露细胞中纤毛的丢失和鳞状分化的增加。免疫荧光染色证实了这些现象。在二乙酰处理的细胞中也观察到基底细胞角蛋白的过度磷酸化和交联，我们使用平行反应监测来自信地定位和量化角蛋白6中先前未表征的磷酸化位点。总的来说，这些数据确定了上皮中的许多分子变化，这些变化可能对风味诱导的闭塞性细支气管炎的发病机制很重要。更一般地说，这项研究强调了定量蛋白质组学在气道损伤和疾病的体外模型研究中的应用。
Occupational exposures to the diketone flavoring agent, diacetyl, have been associated with bronchiolitis obliterans, a rare condition of airway fibrosis. Model studies in rodents have suggested that the airway epithelium is a major site of diacetyl toxicity, but the effects of diacetyl exposure upon the human airway epithelium are poorly characterized. Here, we performed quantitative LC-MS/MS-based proteomics to study the effects of repeated diacetyl vapor exposures on three-dimensional organotypic cultures of human primary tracheobronchial epithelial cells. Using a label-free approach, we quantified approximately 3,400 proteins and 5,700 phosphopeptides in cell lysates across four independent donors. Altered expression of proteins and phosphopeptides were suggestive of loss of cilia and increased squamous differentiation in diacetyl-exposed cells. These phenomena were confirmed by immunofluorescence staining of culture cross-sections. Hyperphosphorylation, and crosslinking, of basal cell keratins were also observed in diacetyl-treated cells, and we used parallel reaction monitoring to confidently localize and quantify previously uncharacterized sites of phosphorylation in keratin 6. Collectively, these data identify numerous molecular changes in the epithelium that may be important to the pathogenesis of flavoring-induced bronchiolitis obliterans. More generally, this study highlights the utility of quantitative proteomics for the study of in vitro models of airway injury and disease.
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影响因子：
3.7
作者：
Hackett NR;Shaykhiev R;Walters MS;Wang R;Zwick RK;Ferris B;Witover B;Salit J;Crystal RG
通讯作者：
Crystal RG
影响因子：
10.4
作者：
Allen JG;Flanigan SS;LeBlanc M;Vallarino J;MacNaughton P;Stewart JH;Christiani DC
通讯作者：
Christiani DC
DOI：
10.1165/rcmb.2011-0353oc
发表时间：
2012-07-01
影响因子：
6.4
作者：
Herfs, Michael;Hubert, Pascale;Delvenne, Philippe
通讯作者：
Delvenne, Philippe
影响因子：
10.4
作者：
ECKERT, RL;RORKE, EA
通讯作者：
RORKE, EA
DOI：
10.1165/rcmb.2014-0483oc
发表时间：
2015-10-01
影响因子：
6.4
作者：
Frommer, Adrien;Hjeij, Rim;Omran, Heymut
通讯作者：
Omran, Heymut