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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
- 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.
- Subjects :
- Male
0301 basic medicine
Adolescent
Proteome
Primary Cell Culture
Cell Culture Techniques
Bronchiolitis obliterans
Diacetyl
Respiratory Mucosa
Biology
Biochemistry
Article
Cornified envelope
Young Adult
03 medical and health sciences
chemistry.chemical_compound
0302 clinical medicine
Keratin
medicine
Humans
Cilia
Phosphorylation
chemistry.chemical_classification
Keratin-6
Cell Differentiation
Epithelial Cells
Molecular Sequence Annotation
General Chemistry
Middle Aged
medicine.disease
Molecular biology
Epithelium
Squamous metaplasia
Flavoring Agents
Gene Ontology
030104 developmental biology
medicine.anatomical_structure
Gene Expression Regulation
chemistry
030220 oncology & carcinogenesis
Respiratory epithelium
Volatilization
Toxicant
Subjects
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