title: Decreased fibronectin production significantly contributes to dysregulated repair of asthmatic epithelium.
abstract: BACKGROUND
  Damage to airway epithelium is followed by deposition of extracellular matrix (ECM) and migration of adjacent epithelial cells. We have shown that epithelial cells from children with asthma fail to heal a wound in vitro.
  
  
  OBJECTIVE
  To determine whether dysregulated ECM production by the epithelium plays a role in aberrant repair in asthma.
  
  
  METHODS
  Airway epithelial cells (AEC) from children with asthma (n = 36), healthy atopic control subjects (n = 23), and healthy nonatopic control subjects (n = 53) were investigated by microarray, gene expression and silencing, transcript regulation analysis, and ability to close mechanical wounds.
  
  
  RESULTS
  Time to repair a mechanical wound in vitro by AEC from healthy and atopic children was not significantly different and both were faster than AEC from children with asthma. Microarray analysis revealed differential expression of multiple gene sets associated with repair and remodeling in asthmatic AEC. Fibronectin (FN) was the only ECM component whose expression was significantly lower in asthmatic AEC. Expression differences were verified by quantitative polymerase chain reaction and ELISA, and reduced FN expression persisted in asthmatic cells over passage. Silencing of FN expression in nonasthmatic AEC inhibited wound repair, whereas addition of FN to asthmatic AEC restored reparative capacity. Asthmatic AEC failed to synthesize FN in response to wounding or cytokine/growth factor stimulation. Exposure to 5', 2'deoxyazacytidine had no effect on FN expression and subsequent analysis of the FN promoter did not show evidence of DNA methylation.
  
  
  CONCLUSIONS
  These data show that the reduced capacity of asthmatic epithelial cells to secrete FN is an important contributor to the dysregulated AEC repair observed in these cells.
authors.affiliation: Department of Respiratory Medicine, Princess Margaret Hospital for Children, Perth, 6001, Western Australia, Australia. anthonyk@ichr.uwa.edu.au
authors.forename: Anthony
authors.initials: A
authors.lastname: Kicic
authors.forename: Teal S
authors.initials: TS
authors.lastname: Hallstrand
authors.forename: Erika N
authors.initials: EN
authors.lastname: Sutanto
authors.forename: Paul T
authors.initials: PT
authors.lastname: Stevens
authors.forename: Michael S
authors.initials: MS
authors.lastname: Kobor
authors.forename: Christopher
authors.initials: C
authors.lastname: Taplin
authors.forename: Peter D
authors.initials: PD
authors.lastname: Paré
authors.forename: Richard P
authors.initials: RP
authors.lastname: Beyer
authors.forename: Stephen M
authors.initials: SM
authors.lastname: Stick
authors.forename: Darryl A
authors.initials: DA
authors.lastname: Knight
mesh_terms: D000293:Adolescent; D001249:Asthma / Q000503:physiopathology*; D001374:Azacitidine / Q000494:pharmacology; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D003513:Cycloheximide / Q000494:pharmacology; D019175:DNA Methylation; D003907:Dexamethasone / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D004847:Epithelial Cells / Q000187:drug effects / Q000502:physiology; D005109:Extracellular Matrix / Q000378:metabolism; D005353:Fibronectins / Q000096:biosynthesis*; D006711:Homocystine / Q000494:pharmacology; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D006967:Hypersensitivity / Q000503:physiopathology; D046228:Microarray Analysis; D020545:Respiratory Mucosa / Q000503:physiopathology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology
chemical_list: D005353:Fibronectins; D006877:Hydroxamic Acids; D053773:Transforming Growth Factor beta1; C012589:trichostatin A; D006711:Homocystine; D003907:Dexamethasone; D003513:Cycloheximide; D001374:Azacitidine; D013755:Tetradecanoylphorbol Acetate