Mice lacking Smad3 show accelerated wound healing and an impaired local inflammatory response

Abstract

The generation of animals lacking SMAD proteins, which transduce signals from transforming growth factor-β (TGF-β), has made it possible to explore the contribution of the SMAD proteins to TGF-β activity in vivo. Here we report that, in contrast to predictions made on the basis of the ability of exogenous TGF-β to improve wound healing, Smad3-null (Smad3ex8/ex8) mice paradoxically show accelerated cutaneous wound healing compared with wild-type mice, characterized by an increased rate of re-epithelialization and significantly reduced local infiltration of monocytes. Smad3ex8/ex8 keratinocytes show altered patterns of growth and migration, and Smad3ex8/ex8 monocytes exhibit a selectively blunted chemotactic response to TGF-β. These data are, to our knowledge, the first to implicate Smad3 in specific pathways of tissue repair and in the modulation of keratinocyte and monocyte function in vivo.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1: Smad3 disruption accelerates cutaneous wound healing.
Figure 2: Accelerated wound healing in Smad3-null mice is associated with a reduced monocytic response.
Figure 3: Addition of TGF-β1 to Smad3–/– wounds has no effect on re-epithelialization but enhances matrix production.
Figure 4: Smad3 is required for TGF-β-induced monocyte chemotaxis and TGF-β expression.
Figure 5: Smad3 deletion modulates keratinocyte proliferation and migration.

References

  1. 1

    Massague, J. TGF-beta signal transduction. Annu. Rev. Biochem. 67, 753–791 (1998).

    CAS  Article  Google Scholar 

  2. 2

    Derynck, R., Zhang, Y. & Feng, X. H. Smads: transcriptional activators of TGF-beta responses. Cell 95, 737–740 (1998).

    CAS  Article  Google Scholar 

  3. 3

    Roberts, A. B. Transforming growth factor-β: activity and efficacy in animal models of wound healing. Wound Repair Regen. 3, 408–418 (1995).

    CAS  Article  Google Scholar 

  4. 4

    O"Kane, S. & Ferguson, M. W. J. Transforming growth factor beta s and wound healing. Int. J. Biochem. Cell Biol. 29, 63–78 (1997).

    CAS  Article  Google Scholar 

  5. 5

    Yang, X. et al. Targeted disruption of SMAD3 results in impaired mucosal immunity and diminished T cell responsiveness to TGF-beta. EMBO J. 18, 1280–1291 (1999).

    CAS  Article  Google Scholar 

  6. 6

    Datto, M. B. et al. Targeted disruption of Smad3 reveals an essential role in transforming growth factor beta-mediated signal transduction. Mol. Cell Biol. 19, 2495–2504 (1999).

    CAS  Article  Google Scholar 

  7. 7

    Zhu, Y., Richardson, J. A., Parada, L. F., & Graff, J. M. Smad3 mutant mice develop metastatic colorectal cancer. Cell 18, 703–714 (1998).

    Article  Google Scholar 

  8. 8

    Weinstein, M., Yang, X., Li, C., Xu, X., & Deng, C. Failure of extraembryonic membrane formation and mesoderm induction in embryos lacking the tumor suppressor Smad2. Proc. Natl Acad. Sci. USA 95, 9378–9383 (1998).

    CAS  Article  Google Scholar 

  9. 9

    Ashcroft, G. S. et al. Estrogen accelerates cutaneous wound healing associated with an increase in TGF-beta1 levels. Nature Med. 3, 1209–1215 (1997).

    CAS  Article  Google Scholar 

  10. 10

    Gross, J. et al. On the mechanism of skin wound “contraction”: a granulation tissue “knockout” with a normal phenotype. Proc. Natl Acad. Sci. USA 92, 5982–5986 (1995).

    CAS  Article  Google Scholar 

  11. 11

    Wahl, S. M. et al. Transforming growth factor type beta induces monocyte chemotaxis and growth factor production. Proc. Natl Acad. Sci. USA 84, 5788–5792 (1987).

    CAS  Article  Google Scholar 

  12. 12

    Leibovich, S. J. & Ross, R. The role of the macrophage in wound repair. A study with hydrocortisone and antimacrophage serum. Am. J. Pathol. 78, 71–100 (1975).

    CAS  PubMed  PubMed Central  Google Scholar 

  13. 13

    McCartney-Francis, N., & Wahl, S. M. Transforming growth factor beta: a matter of life and death. J. Leuk. Biol. 55, 401–409 (1994).

    CAS  Article  Google Scholar 

  14. 14

    Pierce, G. F. et al. Transforming growth factor beta reverses the glucocorticoid-induced wound-healing deficit in rats: possible regulation in macrophages by platelet-derived growth factor. Proc. Natl Acad. Sci. USA 86, 2229–2233 (1989).

    CAS  Article  Google Scholar 

  15. 15

    Vindevoghel, L. et al. SMAD3/4-dependent transcriptional activation of the human type VII collagen gene (COL7A1) promoter by transforming growth factor beta. Proc. Natl Acad. Sci. USA 95, 14769–14774 (1998).

    CAS  Article  Google Scholar 

  16. 16

    Chen, S. J. et al. Stimulation of type I collagen transcription in human skin fibroblasts by TGF-beta: involvement of Smad3. J. Invest. Dermatol. 112, 49–57 (1999).

    CAS  Article  Google Scholar 

  17. 17

    Hocevar, B. A., Brown, T. L. & Howe, P. H. TGF-beta induces fibronectin synthesis through a c-Jun N-terminal kinase-dependent, Smad4-independent pathway. EMBO J. 18, 1345–1356 (1999).

    CAS  Article  Google Scholar 

  18. 18

    Wiseman, D. M., Polverini, P. J., Kamp, D. W. & Leibovich, S. J. Transforming growth factor-beta (TGF beta) is chemotactic for human monocytes and induces their expression of angiogenic activity. Biochem. Biophys. Res. Commun. 157, 793–800 (1988).

    CAS  Article  Google Scholar 

  19. 19

    Wahl, S. M., Allen, J. B., Weeks, B. S., Wong, H. L. & Klotman, P. E. Transforming growth factor beta enhances integrin expression and type IV collagenase secretion in human monocytes. Proc. Natl Acad. Sci. USA 90, 4577–4581 (1993).

    CAS  Article  Google Scholar 

  20. 20

    Zambruno, G. et al. Transforming growth factor-beta 1 modulates beta 1 and beta 5 integrin receptors and induces the de novo expression of the alpha v beta 6 heterodimer in normal human keratinocytes: implications for wound healing. J. Cell Biol. 129, 853–865 (1995).

    CAS  Article  Google Scholar 

  21. 21

    Mustoe, T. A., Pierce, G. F., Morishima, C. & Deuel, T. F. Growth factor-induced acceleration of tissue repair through direct and inductive activities in a rabbit dermal ulcer model. J. Clin. Invest. 87, 694–703 (1991).

    CAS  Article  Google Scholar 

  22. 22

    Hebda, P. A. Stimulatory effects of transforming growth factor-beta and epidermal growth factor on epidermal cell outgrowth from porcine skin explant cultures. J. Invest. Dermatol. 91, 440–445 (1988).

    CAS  Article  Google Scholar 

  23. 23

    Yanagisawa, K. et al. Induction of apoptosis by Smad3 and down-regulation of Smad3 expression in response to TGF-beta in human normal lung epithelial cells. Oncogene 17, 1743–1747 (1998).

    CAS  Article  Google Scholar 

  24. 24

    Dennler, S., Huet, S. & Gauthier, J. M. A short amino-acid sequence in MH1 domain is responsible for functional differences between Smad2 and Smad3. Oncogene 18, 1643–1648 (1999).

    CAS  Article  Google Scholar 

  25. 25

    Ulloa, L., Doody, J. & Massague, J. Inhibition of transforming growth factor-beta/SMAD signalling by the interferon-gamma/STAT pathway. Nature 397, 710–713 (1999).

    CAS  Article  Google Scholar 

  26. 26

    Yanagisawa, J. et al. Convergence of transforming growth factor-beta and vitamin D signaling pathways on SMAD transcriptional coactivators. Science 283, 1317–1321 (1999).

    CAS  Article  Google Scholar 

  27. 27

    Kurokawa, M. et al. The oncoprotein Evi-1 represses TGF-beta signalling by inhibiting Smad3. Nature 2, 92–96 (1998).

    Article  Google Scholar 

  28. 28

    de Caestecker, M. P. et al. Smad2 transduces common signals from receptor serine-threonine and tyrosine kinases. Genes Dev. 12, 587–592 (1998).

    Article  Google Scholar 

  29. 29

    Kretzschmar, M. et al. A mechanism of repression of TGFbeta/Smad signaling by oncogenic Ras. Genes Dev. 1, 804–816 (1999).

    Article  Google Scholar 

  30. 30

    Liu, X. et al. Transforming growth factor beta-induced phosphorylation of Smad3 is required for growth inhibition and transcriptional induction in epithelial cells. Proc. Natl Acad. Sci. USA 94, 10669–10674 (1997).

    CAS  Article  Google Scholar 

  31. 31

    Feldman, G. et al. STAT5A-deficient mice demonstrate a defect in granulocyte-macrophage colony-stimulating factor-induced proliferation and gene expression. Blood 90, 1768–1776 (1997).

    CAS  PubMed  Google Scholar 

  32. 32

    Dlugosz, A. A., Glick, A. B., Tennenbaum, T., Weinberg, W. C. & Yuspa, S. H. Isolation and utilization of epidermal keratinocytes for oncogene research. Methods Enzymol. 254, 3–20 (1995).

    CAS  Article  Google Scholar 

  33. 33

    Danielpour, D. et al. Immunodetection and quantitation of the two forms of transforming growth factor-beta (TGF-beta 1 and TGF-beta 2) secreted by cells in culture. J. Cell Physiol. 138, 79–86 (1989).

    CAS  Article  Google Scholar 

Download references

Acknowledgements

G.S.A. was supported by a Clinician Scientist Fellowship from the Wellcome Trust. We thank L. Hansen for help with the keratinocyte assays.

Correspondence and requests for materials should be addressed to A.B.R..

Author information

Affiliations

Authors

Corresponding author

Correspondence to Anita B. Roberts.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Ashcroft, G., Yang, X., Glick, A. et al. Mice lacking Smad3 show accelerated wound healing and an impaired local inflammatory response. Nat Cell Biol 1, 260–266 (1999). https://doi.org/10.1038/12971

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/12971

Further reading

Search

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing