title: Adenoviral gene transfer of endothelin-1 in the lung induces pulmonary fibrosis through the activation of focal adhesion kinase.
abstract: Endothelin-1 (ET-1) has been implicated in the development of pulmonary fibrosis, based on its capacity in vitro to promote extracellular matrix (ECM) production and contraction, and on studies showing elevated expression of ET-1 and its receptors in patients with pulmonary fibrosis. However, the in vivo fibrogenic effect of ET-1 is not well characterized. We used the adenoviral-mediated gene transfer of ET-1 to overexpress ET-1 transiently in murine lungs by intratracheal administration. An increased expression of ET-1 for 3 to 10 days after injection resulted in a moderate but reversible fibrotic response, peaking on Day 14 after infection and characterized by the deposition of ECM components, myofibroblast formation, and a significant inflammatory infiltrate, mainly in the peribronchiolar/perivascular region. Adenoviral-mediated ET-1 overexpression activated focal adhesion kinase (FAK) both in vitro, using primary murine lung fibroblasts, and in vivo, intratracheally administered in the lungs of mice. The inhibition of FAK with the compound PF-562,271 prevented ET-1-mediated collagen deposition and myofibroblast formation, thereby preventing the development of lung fibrosis. In conclusion, we demonstrate that the overexpression of ET-1 directly in the lungs of mice can initiate a fibrogenic response characterized by increased ECM deposition and myofibroblast formation, and that this effect of ET-1 can be prevented by inhibition of FAK. Our data suggest that the ET-1/FAK axis may contribute importantly to the pathogenesis of fibrotic disorders, and highlight FAK as a potential therapeutic target in these devastating diseases.
authors.affiliation: Centro de Biología Molecular Severo Ochoa, Consejo Superior de Investigaciones Científicas, Universidad Autónoma de Madrid, Madrid, Spain.
authors.forename: David
authors.initials: D
authors.lastname: Lagares
authors.forename: Oscar
authors.initials: O
authors.lastname: Busnadiego
authors.forename: Rosa Ana
authors.initials: RA
authors.lastname: García-Fernández
authors.forename: Santiago
authors.initials: S
authors.lastname: Lamas
authors.forename: Fernando
authors.initials: F
authors.lastname: Rodríguez-Pascual
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology / Q000473:pathology; D002454:Cell Differentiation; D002478:Cells, Cultured; D019332:Endothelin-1 / Q000096:biosynthesis* / Q000235:genetics; D004789:Enzyme Activation; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D005822:Genetic Vectors; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D008168:Lung / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008810:Mice, Inbred C57BL; D058628:Myofibroblasts / Q000473:pathology / Q000502:physiology; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D011658:Pulmonary Fibrosis / Q000201:enzymology* / Q000276:immunology / Q000473:pathology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D012595:Scleroderma, Systemic / Q000201:enzymology / Q000276:immunology / Q000473:pathology; D013449:Sulfonamides / Q000494:pharmacology; D013601:T-Lymphocytes / Q000276:immunology / Q000473:pathology
chemical_list: D019332:Endothelin-1; D007211:Indoles; C527797:N-methyl-N-(3-((2-(2-oxo-2,3-dihydro-1H-indol-5-ylamino)-5-trifluoromethyl-pyrimidin-4-ylamino)-methyl)-pyridin-2-yl)-methanesulfonamide; D011994:Recombinant Proteins; D013449:Sulfonamides; D051416:Focal Adhesion Protein-Tyrosine Kinases