Idiopathic pulmonary fibrosis is characterized by excessive deposition of collagen in the lung, leading to chronically impaired gas exchange and death1,2,3. Oxidative stress is believed to be critical in this disease pathogenesis4,5,6, although the exact mechanisms remain enigmatic. Protein S-glutathionylation (PSSG) is a post-translational modification of proteins that can be reversed by glutaredoxin-1 (GLRX)7. It remains unknown whether GLRX and PSSG play a role in lung fibrosis. Here, we explored the impact of GLRX and PSSG status on the pathogenesis of pulmonary fibrosis, using lung tissues from subjects with idiopathic pulmonary fibrosis, transgenic mouse models and direct administration of recombinant Glrx to airways of mice with existing fibrosis. We demonstrate that GLRX enzymatic activity was strongly decreased in fibrotic lungs, in accordance with increases in PSSG. Mice lacking Glrx were far more susceptible to bleomycin- or adenovirus encoding active transforming growth factor beta-1 (AdTGFB1)-induced pulmonary fibrosis, whereas transgenic overexpression of Glrx in the lung epithelium attenuated fibrosis. We furthermore show that endogenous GLRX was inactivated through an oxidative mechanism and that direct administration of the Glrx protein into airways augmented Glrx activity and reversed increases in collagen in mice with TGFB1- or bleomycin-induced fibrosis, even when administered to fibrotic, aged animals. Collectively, these findings suggest the therapeutic potential of exogenous GLRX in treating lung fibrosis.
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This work was funded by NIH R35HL135828, R01HL079331, R01HL060014 and R03HL095404 (Y.M.W.J.-H.), an ATS unrestricted grant and NIH R01HL122383 (V.A.), Parker B. Francis fellowships (V.A. and J.L.J.v.d.V.), NIH R01HL05646 (A.v.d.V.), T32HL076122 (D.H.M. and J.T.J.), NCRR 1S10RR019246 (D.J.T.), American Heart Association 16GRNT27660006, NIH R01DK103750 and 1UL1TR001430 (M.M.B.), NIH R01HL133013 and R03 AG051857 (R.M.), P30GM103532 (C.G.I.), FAMRI Young Clinical Scientist Award 113393 and 1K01HL125474-01 (J.L.G.) and U01HL108642 and RC2 HL101715 (N.K.). Analysis of biological specimens and data provided by the Lung Tissue Research Consortium (LTRC) was supported by R03HL095404 (Y.M.W.J.-H.). We would like to thank J. Whitsett (University of Cincinnati) for generously providing the CCSP-rtTA, TetOP-Cre mice, J. Gauldie for providing AdTGFB1, N. Bishop for help with imaging of GLRX and collagen, J. M. Siddesha for technical support and P. Vacek for statistical support.