Inhibition of the αvβ6 integrin leads to limited alteration of TGF-α-induced pulmonary fibrosis

Am J Physiol Lung Cell Mol Physiol. 2014 Apr 15;306(8):L726-35. doi: 10.1152/ajplung.00357.2013. Epub 2014 Feb 7.

Abstract

A number of growth factors and signaling pathways regulate matrix deposition and fibroblast proliferation in the lung. The epidermal growth factor receptor (EGFR) family of receptors and the transforming growth factor-β (TGF-β) family are active in diverse biological processes and are central mediators in the initiation and maintenance of fibrosis in many diseases. Transforming growth factor-α (TGF-α) is a ligand for the EGFR, and doxycycline (Dox)-inducible transgenic mice conditionally expressing TGF-α specifically in the lung epithelium develop progressive fibrosis accompanied with cachexia, changes in lung mechanics, and marked pleural thickening. Although recent studies demonstrate that EGFR activation modulates the fibroproliferative effects involved in the pathogenesis of TGF-β induced pulmonary fibrosis, in converse, the direct role of EGFR induction of the TGF-β pathway in the lung is unknown. The αvβ6 integrin is an important in vivo activator of TGF-β activation in the lung. Immunohistochemical analysis of αvβ6 protein expression and bronchoalveolar analysis of TGF-β pathway signaling indicates activation of the αvβ6/TGF-β pathway only at later time points after lung fibrosis was already established in the TGF-α model. To determine the contribution of the αvβ6/TGF-β pathway on the progression of established fibrotic disease, TGF-α transgenic mice were administered Dox for 4 wk, which leads to extensive fibrosis; these mice were then treated with a function-blocking anti-αvβ6 antibody with continued administration of Dox for an additional 4 wk. Compared with TGF-α transgenic mice treated with control antibody, αvβ6 inhibition significantly attenuated pleural thickening and altered the decline in lung mechanics. To test the effects of genetic loss of the β6 integrin, TGF-α transgenic mice were mated with β6-null mice and the degree of fibrosis was compared in adult mice following 8 wk of Dox administration. Genetic ablation of the β6 integrin attenuated histological and physiological changes in the lungs of TGF-α transgenic mice although a significant degree of fibrosis still developed. In summary, inhibition of the β6 integrin led to a modest, albeit significant, effect on pleural thickening and lung function decline observed with TGF-α-induced pulmonary fibrosis. These data support activation of the αvβ6/TGF-β pathway as a secondary effect contributing to TGF-α-induced pleural fibrosis and suggest a complex contribution of multiple mediators to the maintenance of progressive fibrosis in the lung.

Keywords: TGF-β; epidermal growth factor receptor; integrin; pleural fibrosis; pulmonary fibrosis.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Anti-Bacterial Agents / toxicity
  • Antibodies, Neutralizing
  • Antigens, Neoplasm / genetics
  • Antigens, Neoplasm / metabolism
  • Bronchoalveolar Lavage
  • Collagen
  • Doxycycline / toxicity
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Humans
  • Immunoenzyme Techniques
  • Integrins / antagonists & inhibitors*
  • Integrins / genetics
  • Integrins / metabolism
  • Male
  • Mice
  • Mice, Transgenic
  • Pulmonary Fibrosis / chemically induced
  • Pulmonary Fibrosis / metabolism
  • Pulmonary Fibrosis / pathology*
  • RNA, Messenger / genetics
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transforming Growth Factor alpha / pharmacology*
  • Transforming Growth Factor beta / pharmacology
  • Uteroglobin / physiology

Substances

  • Anti-Bacterial Agents
  • Antibodies, Neutralizing
  • Antigens, Neoplasm
  • Integrins
  • RNA, Messenger
  • Scgb1a1 protein, mouse
  • Transforming Growth Factor alpha
  • Transforming Growth Factor beta
  • integrin alphavbeta6
  • Collagen
  • Uteroglobin
  • Doxycycline