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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

Authors :
Francine L. Kelly
William M. Gwinn
Scott M. Palmer
David M. Brass
J. Will Thompson
Matthew W. Foster
M. Arthur Moseley
Daniel L. Morgan
Ashlee M. Valente
Source :
J Proteome Res
Publication Year :
2017
Publisher :
American Chemical Society (ACS), 2017.

Abstract

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.

Details

ISSN :
15353907 and 15353893
Volume :
16
Database :
OpenAIRE
Journal :
Journal of Proteome Research
Accession number :
edsair.doi.dedup.....333cab01bc458ffcd61c874b5128f0ed
Full Text :
https://doi.org/10.1021/acs.jproteome.6b00672