Decreased Fibronectin Production Significantly Contributes to Dysregulated Repair of Asthmatic Epithelium

• Published in: American journal of respiratory and critical care medicine Vol. 181; no. 9; pp. 889 - 898
• Main Authors: KICIC, Anthony, HALLSTRAND, Teal S, SUTANTO, Erika N, STEVENS, Paul T, KOBOR, Michael S, TAPLIN, Christopher, PARE, Peter D, BEYER, Richard P, STICK, Stephen M, KNIGHT, Darryl A
• Format: Journal Article
• Language: English
• Published: New York, NY American Thoracic Society 01-05-2010
• Subjects: A. Asthma and Allergy; Adolescent; Anesthesia; Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy; Asthma; Asthma - physiopathology; Azacitidine - pharmacology; Biological and medical sciences; Cells, Cultured; Child; Child, Preschool; Cycloheximide - pharmacology; Cytokines; Dexamethasone - pharmacology; DNA Methylation; Enzyme-Linked Immunosorbent Assay; Epithelial Cells - drug effects; Epithelial Cells - physiology; Extracellular matrix; Extracellular Matrix - metabolism; Fibronectins - biosynthesis; Gene expression; Growth factors; Homocysteine; Homocystine - pharmacology; Humans; Hydroxamic Acids - pharmacology; Hypersensitivity - physiopathology; Intensive care medicine; Local anesthesia. Pain (treatment); Medical sciences; Microarray Analysis; Polymerase chain reaction; Respiratory Mucosa - physiopathology; Tetradecanoylphorbol Acetate - pharmacology; Transforming Growth Factor beta1 - pharmacology; Germany; Lung disease; Intensive care; wound repair; Respiratory disease; Bronchus disease; Inflammation; Obstructive pulmonary disease; Epithelium; Wound; Fibronectin; Resuscitation; Asthma
• ISSN: 1073-449X; 1535-4970
• DOI: 10.1164/rccm.200907-1071oc

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.
To determine whether dysregulated ECM production by the epithelium plays a role in aberrant repair in asthma.
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.
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.
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.