Matrix metalloprotease-9 induces transforming growth factor- β₁ production in airway epithelium via activation of epidermal growth factor receptors

Aims: Matrix metalloprotease (MMP)-9 is present in abundance in various chronic airway disorders and is involved in lung remodeling. MMP may cleave membrane-bound precursor proteins and release epidermal growth factor-like ligands that subsequently bind to epidermal growth factor receptor (EGFR). Wehypothesized that MMP-9 may stimulate the airway epithelium to produce fibrogenic mediators through activation of membrane-bound receptors. Main methods: Human airway epithelial cells were grown on air-liquid interface culture inserts. MMP-9 was employed to stimulate the cells. Conditioned medium following MMP-9 stimulation was co-incubated with human lung fibroblasts. Key findings: MMP-9 stimulated human airway epithelial cells to produce transforming growth factor (TGF)-β₁ at both themRNA and protein level. Using amicroarray, increased phosphorylation of EGFR tyrosine kinase (TK) was identified and further confirmed by immunoprecipitation and Western blot analysis. A significant increase in EGF and TGF-α release was observed after MMP-9 had been added for 30min. Protease inhibitor, EGFR monoclonal antibody and EGFR-TK inhibitor blocked this action and subsequent TGF-β₁ production. Neutralizing antibodies against EGF and TGF-α substantially inhibited TGF-β₁ production following MMP-9 stimulation. MMP-9-induced TGF-β₁ production occurred through MAP kinase p44/42 phosphorylation. Selective p44/42 kinase inhibitor UO126 successfully inhibited TGF-β₁ production. Conditioned medium from epithelial cells treated with MMP-9 significantly induced Smad3 phosphorylation and subsequent fibroblast proliferation after 24 h culture. Significance: These data indicate that MMP-9 induces TGF-β₁ production in the airway epithelium through the cleavage of EGF and EGF-like ligands and activating EGFR, suggesting potential targets of therapeutic intervention in airway fibrotic disorders.