title: Integrative proteomic analysis reveals reprograming tumor necrosis factor signaling in epithelial mesenchymal transition.
abstract: The airway epithelium is a semi-impermeable barrier whose disruption by growth factor reprogramming is associated with chronic airway diseases of humans. Transforming growth factor beta (TGFβ)-induced epithelial mesenchymal transition (EMT) plays important roles in airway remodeling characteristic of idiopathic lung fibrosis, asthma and chronic obstructive pulmonary disease (COPD). Inflammation of the airways leads to airway injury and tumor necrosis factor alpha (TNFα) plays an important pro-inflammatory role. Little systematic information about the effects of EMT on TNFα signaling is available. Using an in vitro model of TGFβ-induced EMT in primary human small airway epithelial cells (hSAECs), we applied quantitative proteomics and phosphoprotein profiling to understand the molecular mechanism of EMT and the impact of EMT on innate inflammatory responses. We quantified 7925 proteins and 1348 phosphorylation sites by stable isotope labeling with iTRAQ technology. We found that cellular response to TNFα is cell state dependent and the relative TNFα response in mesenchymal state is highly compressed. Combined bioinformatics analyses of proteome and phosphoproteome indicate that the EMT state is associated with reprogramming of kinome, signaling cascade of upstream transcription regulators, phosphor-networks, and NF-κB dependent cell signaling.
  
  
  
  Epithelial mesenchymal transition and inflammation have important implications for clinical and physiologic manifestations of chronic airway diseases such as severe asthma, COPD, and lung fibrosis. Little systematic information on the interplay between EMT and innate inflammation is available. This study combined quantitative proteomics and phosphorproteomics approach to obtain systems-level insight into the upstream transcription regulators involved in the TGFβ-induced EMT in primary human small airway epithelial cells and to elucidate how EMT impacts on the TNFα signaling pathways. The proteomics and phosphoproteomics analysis indicates that many signaling pathways involved in TGFβ-induced EMT and EMT has profound reprogramming effects on innate inflammation response.
authors.affiliation: Department of Internal Medicine, University of Texas Medical Branch (UTMB), Galveston, TX, United States; Institute for Translational Sciences, UTMB, Galveston, TX, United States; Sealy Center for Molecular Medicine, UTMB, Galveston, TX, United States. Electronic address: yizhao@utmb.edu.
authors.forename: Yingxin
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Department of Internal Medicine, University of Texas Medical Branch (UTMB), Galveston, TX, United States; Institute for Translational Sciences, UTMB, Galveston, TX, United States; Sealy Center for Molecular Medicine, UTMB, Galveston, TX, United States.
authors.forename: Bing
authors.initials: B
authors.lastname: Tian
authors.affiliation: Sealy Center for Molecular Medicine, UTMB, Galveston, TX, United States; Department of Biochemistry & Molecular Biology, UTMB, Galveston, TX, United States.
authors.forename: Rovshan G
authors.initials: RG
authors.lastname: Sadygov
authors.affiliation: Department of Internal Medicine, University of Texas Medical Branch (UTMB), Galveston, TX, United States.
authors.forename: Yueqing
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Internal Medicine, University of Texas Medical Branch (UTMB), Galveston, TX, United States; Institute for Translational Sciences, UTMB, Galveston, TX, United States; Sealy Center for Molecular Medicine, UTMB, Galveston, TX, United States. Electronic address: arbrasie@utmb.edu.
authors.forename: Allan R
authors.initials: AR
authors.lastname: Brasier
mesh_terms: D056151:Airway Remodeling; D056809:Alveolar Epithelial Cells / Q000378:metabolism; D002461:Cell Line, Transformed; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects*; D006801:Humans; D007249:Inflammation / Q000209:etiology; D010750:Phosphoproteins / Q000032:analysis; D040901:Proteomics / Q000379:methods*; D015398:Signal Transduction; D016212:Transforming Growth Factor beta / Q000494:pharmacology*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*
chemical_list: D010750:Phosphoproteins; D016212:Transforming Growth Factor beta; D014409:Tumor Necrosis Factor-alpha
keywords: Epithelial mesenchymal transition; Innate immune response; Mass spectrometry; Phosphoproteomics; Proteomics