Proteomic Analysis of Primary Human Airway Epithelial Cells Exposed to the Respiratory Toxicant Diacetyl

Details

• Personal Author:
  Brass DM ; Foster MW ; Gwinn WM ; Kelly FL ; Morgan DL ; Moseley MA ; Palmer SM ; Thompson JW ; Valente AM
• Description:
  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 3D organotypic cultures of human primary tracheobronchial epithelial cells. Using a label-free approach, we quantified approximately 3400 proteins and 5700 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 cross-linking of basal cell keratins were also observed in diacetyltreated 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. [Description provided by NIOSH]
• Subjects:
  Animals Biomarkers Diacetyl Environmental Exposure Fibrosis Flavoring Agents Hazardous Substances Laboratories Lung Lung Diseases Occupational Health Respiratory System Respiratory Tract Diseases Risk Assessment Risk Factors Safety Workplace

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• Keywords:
  Author Keywords: BO Bronchiolitis Butanedione Butter Flavoring Cell Biology Cellular Function Cellular Reactions Cornified Envelope Diseases Exposure Levels Flavorings Laboratory Animals Lung Disease Lung Disorders Lung Fibrosis Pentanedione PRM Proteins Repetin Risk Assessment Risk Factors RSPH4A Squamous Metaplasia TGM1 Toxic Effects Toxins Work Environment Workers

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• ISSN:
  1535-3893
• Document Type:
  Text
• Funding:
  Grant
• Genre:
  Journal Article
• Place as Subject:
  North Carolina ; OSHA Region 4
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Volume:
  16
• Issue:
  2
• NIOSHTIC Number:
  nn:20052070
• Citation:
  J Proteome Res 2016 Dec; 16(2):538-549
• Contact Point Address:
  Matthew W. Foster, Department of Medicine and Duke Proteomics and Metabolomics Shared Resource, Duke University Medical Center, Durham, North Carolina 27710
• CAS Registry Number:
  Butanedione (CAS RN 431-03-8)
• Federal Fiscal Year:
  2017
• Performing Organization:
  Duke University
• Peer Reviewed:
  True
• Start Date:
  20140901
• Source Full Name:
  Journal of Proteome Research
• End Date:
  20160831
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:65a316c8603e33a92302ae832a0522c65e4bbc6d6ce1a958281d5718c4c691ff536aac95cf129d6c63d3c3519d39f0e2206cbc7ada8369c91ffacc77daa4ecd5
• Download URL:
  https://stacks.cdc.gov/view/cdc/211670/cdc_211670_DS1.pdf
• File Type:
  [PDF - 5.18 MB ]