ADAM10-mediated ephrin-B2 shedding promotes myofibroblast activation and organ fibrosis

  • A Corrigendum to this article was published on 01 December 2017

Abstract

Maladaptive wound healing responses to chronic tissue injury result in organ fibrosis. Fibrosis, which entails excessive extracellular matrix (ECM) deposition and tissue remodeling by activated myofibroblasts, leads to loss of proper tissue architecture and organ function; however, the molecular mediators of myofibroblast activation have yet to be fully identified. Here we identify soluble ephrin-B2 (sEphrin-B2) as a new profibrotic mediator in lung and skin fibrosis. We provide molecular, functional and translational evidence that the ectodomain of membrane-bound ephrin-B2 is shed from fibroblasts into the alveolar airspace after lung injury. Shedding of sEphrin-B2 promotes fibroblast chemotaxis and activation via EphB3 and/or EphB4 receptor signaling. We found that mice lacking ephrin-B2 in fibroblasts are protected from skin and lung fibrosis and that a disintegrin and metalloproteinase 10 (ADAM10) is the major ephrin-B2 sheddase in fibroblasts. ADAM10 expression is increased by transforming growth factor (TGF)-β1, and ADAM10-mediated sEphrin-B2 generation is required for TGF-β1-induced myofibroblast activation. Pharmacological inhibition of ADAM10 reduces sEphrin-B2 levels in bronchoalveolar lavage and prevents lung fibrosis in mice. Consistent with the mouse data, ADAM10–sEphrin-B2 signaling is upregulated in fibroblasts from human subjects with idiopathic pulmonary fibrosis. These results uncover a new molecular mechanism of tissue fibrogenesis and identify sEphrin-B2, its receptors EphB3 and EphB4 and ADAM10 as potential therapeutic targets in the treatment of fibrotic diseases.

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Figure 1: Bleomycin-induced lung fibrosis is dependent on ephrin-B2 in fibroblasts.
Figure 2: Ephrin-B2 ectodomain is shed by fibroblasts upon lung injury.
Figure 3: The ephrin-B2 ectodomain directs fibroblast migration, invasion and myofibroblast differentiation in vitro and in vivo.
Figure 4: ADAM10-mediated sEphrin-B2 generation is required for TGF-β1-induced myofibroblast differentiation.
Figure 5: ADAM10 inhibition prevents ephrin-B2 shedding, myofibroblast formation and lung fibrosis in mice.
Figure 6: ADAM10-sEphrin-B2 signaling is upregulated in IPF.

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  • 20 November 2017

    In the version of this article initially published online, the positions of the colored boxes in the key of Figure 5f were inverted. The treatment group is represented by the red line of the graph and the control group by the blue line. The error has been corrected in the print, PDF and HTML versions of this article.

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Acknowledgements

Authors would like to thank P. Datta, S. Nakamura, H. Endisha and J. Rockel (all from the University Health Network) for their technical assistance with mouse breeding and genotyping. The authors gratefully acknowledge funding support by University of Montreal Hospital Research Centre and University of Montreal (M.K.); Campaign to Cure Arthritis via the Toronto General and Western Foundation, University Health Network, Toronto (M.K.); an American Thoracic Society Foundation and Pulmonary Fibrosis Foundation Research Grant and the Marie A. Coyle Research Grant from the Scleroderma Foundation (D.L.), and by the National Institutes of Health, HL108975 and a grant from the Scleroderma Research Foundation (A.M.T).

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D.L. designed most of the experiments, performed in vitro and in vivo mouse experiments, analyzed the data and generated the figures. P.G.-K. and M.B. were involved in the generation of Ephrinb2-CKO mice. A.S., P.G., N.A. and D.M.S. performed and analyzed in vitro experiments related to the ADAM10–ephrin-B2–EphB3/4 pathway in fibroblasts. C.K.P. and A.F. performed in vivo studies with ADAM10 inhibitor. C.T. was involved in histological characterization of mouse experiments in skin fibrosis model. M.S., A.P., S.B.M., R.K., K.E.B. and B.S.S. provided human lung fibroblasts, plasma and bronchoalveolar lavage fluid from individuals with IPF and healthy controls. M.B. and R.G. provided intellectual input on project design and troubleshooting. B.W. performed protein expression studies in mouse samples. J.W., H.F., J.-P.P. and J.M.-P. were involved in the characterization of mouse phenotype and troubleshooting with experiments related to ephrin biology. M.K. designed the original concept and led the entire team during the course of this study. D.L., A.M.T. and M.K. designed the study experiments, supervised the project and took overall responsibility for writing the manuscript with the help of all the authors.

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Correspondence to David Lagares or Mohit Kapoor.

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Lagares, D., Ghassemi-Kakroodi, P., Tremblay, C. et al. ADAM10-mediated ephrin-B2 shedding promotes myofibroblast activation and organ fibrosis. Nat Med 23, 1405–1415 (2017). https://doi.org/10.1038/nm.4419

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