Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Lineage tracing and genetic ablation of ADAM12+ perivascular cells identify a major source of profibrotic cells during acute tissue injury

Nature Medicine volume 18, pages 1262–1270 (2012)Cite this article

Subjects

Abstract

Profibrotic cells that develop upon injury generate permanent scar tissue and impair organ recovery, though their origin and fate are unclear. Here we show that transient expression of ADAM12 (a disintegrin and metalloprotease 12) identifies a distinct proinflammatory subset of platelet-derived growth factor receptor-α–positive stromal cells that are activated upon acute injury in the muscle and dermis. By inducible genetic fate mapping, we demonstrate in vivo that injury-induced ADAM12+ cells are specific progenitors of a major fraction of collagen-overproducing cells generated during scarring, which are progressively eliminated during healing. Genetic ablation of ADAM12+ cells, or knockdown of ADAM12, is sufficient to limit generation of profibrotic cells and interstitial collagen accumulation. ADAM12+ cells induced upon injury are developmentally distinct from muscle and skin lineage cells and are derived from fetal ADAM12+ cells programmed during vascular wall development. Thus, our data identify injury-activated profibrotic progenitors residing in the perivascular space that can be targeted through ADAM12 to limit tissue scarring.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: ADAM12 expression identifies a proinflammatory subset of gp38+PDGFR-α+ stromal cells transiently induced upon dermis and muscle injury.
Figure 2: The majority of myofibroblasts developing in vivo upon injury are progeny of ADAM12+ cells.
Figure 3: ADAM12+ profibrotic progenitors are induced de novo by tissue injury.
Figure 4: Ablation of ADAM12+ cells induced de novo by injury reduces inflammation and tissue fibrosis.
Figure 5: Tissue injury reactivates ADAM12+ cells programmed during nerve and vascular wall development.
Figure 6: Mesenchymal perivascular ADAM12+ cells are profibrotic progenitors.

Similar content being viewed by others

Article 01 December 2022

Zixuan Zhao, Xinyi Chen, … Hanry Yu

References

  1. Collins, C.A. et al. Stem cell function, self-renewal, and behavioral heterogeneity of cells from the adult muscle satellite cell niche. Cell 122, 289–301 (2005).

    Article  CAS  PubMed  Google Scholar 

  2. Blanpain, C. & Fuchs, E. Epidermal homeostasis: a balancing act of stem cells in the skin. Nat. Rev. Mol. Cell Biol. 10, 207–217 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Wynn, T.A. Cellular and molecular mechanisms of fibrosis. J. Pathol. 214, 199–210 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Brack, A.S. et al. Increased Wnt signaling during aging alters muscle stem cell fate and increases fibrosis. Science 317, 807–810 (2007).

    Article  CAS  PubMed  Google Scholar 

  5. Li, Y. et al. Transforming growth factor-β1 induces the differentiation of myogenic cells into fibrotic cells in injured skeletal muscle: a key event in muscle fibrogenesis. Am. J. Pathol. 164, 1007–1019 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Uezumi, A., Fukada, S., Yamamoto, N., Takeda, S. & Tsuchida, K. Mesenchymal progenitors distinct from satellite cells contribute to ectopic fat cell formation in skeletal muscle. Nat. Cell Biol. 12, 143–152 (2010).

    Article  CAS  PubMed  Google Scholar 

  7. Joe, A.W. et al. Muscle injury activates resident fibro/adipogenic progenitors that facilitate myogenesis. Nat. Cell Biol. 12, 153–163 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Olson, L.E. & Soriano, P. Increased PDGFRα activation disrupts connective tissue development and drives systemic fibrosis. Dev. Cell 16, 303–313 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Herzog, E.L. & Bucala, R. Fibrocytes in health and disease. Exp. Hematol. 38, 548–556 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Hinz, B. et al. The myofibroblast: one function, multiple origins. Am. J. Pathol. 170, 1807–1816 (2007).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Crisan, M. et al. A perivascular origin for mesenchymal stem cells in multiple human organs. Cell Stem Cell 3, 301–313 (2008).

    Article  CAS  PubMed  Google Scholar 

  12. Real, C., Glavieux-Pardanaud, C., Vaigot, P., Le-Douarin, N. & Dupin, E. The instability of the neural crest phenotypes: Schwann cells can differentiate into myofibroblasts. Int. J. Dev. Biol. 49, 151–159 (2005).

    Article  CAS  PubMed  Google Scholar 

  13. Thiery, J.P., Acloque, H., Huang, R.Y. & Nieto, M.A. Epithelial-mesenchymal transitions in development and disease. Cell 139, 871–890 (2009).

    Article  CAS  PubMed  Google Scholar 

  14. Joyner, A.L. & Zervas, M. Genetic inducible fate mapping in mouse: establishing genetic lineages and defining genetic neuroanatomy in the nervous system. Dev. Dyn. 235, 2376–2385 (2006).

    Article  PubMed  Google Scholar 

  15. Horiuchi, K. et al. Substrate selectivity of epidermal growth factor-receptor ligand sheddases and their regulation by phorbol esters and calcium influx. Mol. Biol. Cell 18, 176–188 (2007).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Shi, Z., Xu, W., Loechel, F., Wewer, U.M. & Murphy, L.J. ADAM 12, a disintegrin metalloprotease, interacts with insulin-like growth factor–binding protein-3. J. Biol. Chem. 275, 18574–18580 (2000).

    Article  CAS  PubMed  Google Scholar 

  17. Atfi, A. et al. The disintegrin and metalloproteinase ADAM12 contributes to TGF-β signaling through interaction with the type II receptor. J. Cell Biol. 178, 201–208 (2007).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Kurisaki, T., Masuda, A., Osumi, N., Nabeshima, Y. & Fujisawa-Sehara, A. Spatially- and temporally-restricted expression of meltrin α (ADAM12) and β (ADAM19) in mouse embryo. Mech. Dev. 73, 211–215 (1998).

    Article  CAS  PubMed  Google Scholar 

  19. Le Pabic, H. et al. ADAM12 in human liver cancers: TGF-β–regulated expression in stellate cells is associated with matrix remodeling. Hepatology 37, 1056–1066 (2003).

    Article  CAS  PubMed  Google Scholar 

  20. Borneman, A., Kuschel, R. & Fujisawa-Sehara, A. Analysis for transcript expression of meltrin α in normal, regenerating, and denervated rat muscle. J. Muscle Res. Cell Motil. 21, 475–480 (2000).

    Article  CAS  PubMed  Google Scholar 

  21. Shi-Wen, X. et al. Endogenous endothelin-1 signaling contributes to type I collagen and CCN2 overexpression in fibrotic fibroblasts. Matrix Biol. 26, 625–632 (2007).

    Article  PubMed  Google Scholar 

  22. Skubitz, K.M. & Skubitz, A.P. Gene expression in aggressive fibromatosis. J. Lab. Clin. Med. 143, 89–98 (2004).

    Article  CAS  PubMed  Google Scholar 

  23. Peduto, L. et al. ADAM12 is highly expressed in carcinoma-associated stroma and is required for mouse prostate tumor progression. Oncogene 25, 5462–5466 (2006).

    Article  CAS  PubMed  Google Scholar 

  24. Peduto, L. et al. Inflammation recapitulates the ontogeny of lymphoid stromal cells. J. Immunol. 182, 5789–5799 (2009).

    Article  CAS  PubMed  Google Scholar 

  25. Harris, J.B. Myotoxic phospholipases A2 and the regeneration of skeletal muscles. Toxicon 42, 933–945 (2003).

    Article  CAS  PubMed  Google Scholar 

  26. Jessen, K.R. & Mirsky, R. Negative regulation of myelination: relevance for development, injury, and demyelinating disease. Glia 56, 1552–1565 (2008).

    Article  PubMed  Google Scholar 

  27. Hernandez-Gea, V. & Friedman, S.L. Pathogenesis of liver fibrosis. Annu. Rev. Pathol. 6, 425–456 (2011).

    Article  CAS  PubMed  Google Scholar 

  28. Passino, M.A., Adams, R.A., Sikorski, S.L. & Akassoglou, K. Regulation of hepatic stellate cell differentiation by the neurotrophin receptor p75NTR. Science 315, 1853–1856 (2007).

    Article  CAS  PubMed  Google Scholar 

  29. Gaengel, K., Genove, G., Armulik, A. & Betsholtz, C. Endothelial-mural cell signaling in vascular development and angiogenesis. Arterioscler. Thromb. Vasc. Biol. 29, 630–638 (2009).

    Article  CAS  PubMed  Google Scholar 

  30. Luche, H., Weber, O., Nageswara Rao, T., Blum, C. & Fehling, H.J. Faithful activation of an extra-bright red fluorescent protein in “knock-in” Cre-reporter mice ideally suited for lineage tracing studies. Eur. J. Immunol. 37, 43–53 (2007).

    Article  CAS  PubMed  Google Scholar 

  31. Schönig, K., Schwenk, F., Rajewsky, K. & Bujard, H. Stringent doxycycline dependent control of CRE recombinase in vivo. Nucleic Acids Res. 30, e134 (2002).

    Article  PubMed  PubMed Central  Google Scholar 

  32. Srinivas, S. et al. Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus. BMC Dev. Biol. 1, 4 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Le Douarin, N.M., Calloni, G.W. & Dupin, E. The stem cells of the neural crest. Cell Cycle 7, 1013–1019 (2008).

    Article  CAS  PubMed  Google Scholar 

  34. Verrecchia, F. & Mauviel, A. Transforming growth factor-β and fibrosis. World J. Gastroenterol. 13, 3056–3062 (2007).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Betancur, P., Bronner-Fraser, M. & Sauka-Spengler, T. Assembling neural crest regulatory circuits into a gene regulatory network. Annu. Rev. Cell Dev. Biol. 26, 581–603 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Jessen, K.R. & Mirsky, R. The origin and development of glial cells in peripheral nerves. Nat. Rev. Neurosci. 6, 671–682 (2005).

    Article  CAS  PubMed  Google Scholar 

  37. Buchstaller, J. et al. Efficient isolation and gene expression profiling of small numbers of neural crest stem cells and developing Schwann cells. J. Neurosci. 24, 2357–2365 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Joseph, N.M. et al. Neural crest stem cells undergo multilineage differentiation in developing peripheral nerves to generate endoneurial fibroblasts in addition to Schwann cells. Development 131, 5599–5612 (2004).

    Article  CAS  PubMed  Google Scholar 

  39. Danielian, P.S., Muccino, D., Rowitch, D.H., Michael, S.K. & McMahon, A.P. Modification of gene activity in mouse embryos in utero by a tamoxifen-inducible form of Cre recombinase. Curr. Biol. 8, 1323–1326 (1998).

    Article  CAS  PubMed  Google Scholar 

  40. Pittenger, M.F. et al. Multilineage potential of adult human mesenchymal stem cells. Science 284, 143–147 (1999).

    Article  CAS  PubMed  Google Scholar 

  41. Morikawa, S. et al. Prospective identification, isolation, and systemic transplantation of multipotent mesenchymal stem cells in murine bone marrow. J. Exp. Med. 206, 2483–2496 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Iredale, J.P. Models of liver fibrosis: exploring the dynamic nature of inflammation and repair in a solid organ. J. Clin. Invest. 117, 539–548 (2007).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Griffin, J.W. & Thompson, W.J. Biology and pathology of nonmyelinating Schwann cells. Glia 56, 1518–1531 (2008).

    Article  PubMed  Google Scholar 

  44. da Silva Meirelles, L., Caplan, A.I. & Nardi, N.B. In search of the in vivo identity of mesenchymal stem cells. Stem Cells 26, 2287–2299 (2008).

    Article  PubMed  Google Scholar 

  45. Olson, L.E. & Soriano, P. PDGFRβ signaling regulates mural cell plasticity and inhibits fat development. Dev. Cell 20, 815–826 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Chung, A.S., Lee, J. & Ferrara, N. Targeting the tumour vasculature: insights from physiological angiogenesis. Nat. Rev. Cancer 10, 505–514 (2010).

    Article  CAS  PubMed  Google Scholar 

  47. Lin, S.L., Kisseleva, T., Brenner, D.A. & Duffield, J.S. Pericytes and perivascular fibroblasts are the primary source of collagen-producing cells in obstructive fibrosis of the kidney. Am. J. Pathol. 173, 1617–1627 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Humphreys, B.D. et al. Fate tracing reveals the pericyte and not epithelial origin of myofibroblasts in kidney fibrosis. Am. J. Pathol. 176, 85–97 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Göritz, C. et al. A pericyte origin of spinal cord scar tissue. Science 333, 238–242 (2011).

    Article  PubMed  Google Scholar 

  50. Jørgensen, L.H., Jensen, C.H., Wewer, U.M. & Schroder, H.D. Transgenic overexpression of ADAM12 suppresses muscle regeneration and aggravates dystrophy in aged mdx mice. Am. J. Pathol. 171, 1599–1607 (2007).

    Article  PubMed  PubMed Central  Google Scholar 

  51. Blobel, C.P. ADAMs: key components in EGFR signalling and development. Nat. Rev. Mol. Cell Biol. 6, 32–43 (2005).

    Article  CAS  PubMed  Google Scholar 

  52. Solomon, E., Li, H., Muggy, S.D., Syta, E. & Zolkiewska, A. The role of SnoN in transforming growth factor β1-induced expression of metalloprotease-disintegrin ADAM12. J. Biol. Chem. 285, 21969–21977 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  53. Takashima, Y. et al. Neuroepithelial cells supply an initial transient wave of MSC differentiation. Cell 129, 1377–1388 (2007).

    Article  CAS  PubMed  Google Scholar 

  54. Le Douarin, N.M., Brito, J.M. & Creuzet, S. Role of the neural crest in face and brain development. Brain Res. Rev. 55, 237–247 (2007).

    Article  PubMed  Google Scholar 

  55. Sparwasser, T., Gong, S., Li, J.Y.H. & Eberl, G. General method for the modification of different BAC types and the rapid generation of BAC transgenic mice. Genesis 38, 39–50 (2004).

    Article  CAS  PubMed  Google Scholar 

  56. Arsic, N. et al. Vascular endothelial growth factor stimulates skeletal muscle regeneration in vivo. Mol. Ther. 10, 844–854 (2004).

    Article  CAS  PubMed  Google Scholar 

  57. Pappenheimer, A.M. Jr., Harper, A.A., Moynihan, M. & Brockes, J.P. Diphtheria toxin and related proteins: effect of route of injection on toxicity and the determination of cytotoxicity for various cultured cells. J. Infect. Dis. 145, 94–102 (1982).

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was funded by the Institut Pasteur, grants from the Mairie de Paris, the Agence Nationale de la Recherche and an Excellence grant from the European Commission. S.E.D.C. is funded by La Ligue contre le Cancer. We thank H.-J. Fehling (University Clinics Ulm, Ulm, Germany) for providing the Rosa26floxSTOP-RFP mice, H. Bujard (ZMBH, Heidelberg, Germany) for providing the LC-1 mice and A. Farr (University of Washington, Seattle) for gp38-specific antibody. We thank L. Polomack and the team of the Centre d'Ingénierie Génétique Murine for technical assistance and members of the Development of Lymphoid Tissue Unit for discussions.

Author information

Authors and Affiliations

  1. Institut Pasteur, Lymphoid Tissue Development Unit, Paris, France

    Sophie Dulauroy, Selene E Di Carlo, Gérard Eberl & Lucie Peduto

  2. Centre National de la Recherche Scientifique (CNRS), Unité de Recherche Associée (URA)1961, Paris, France

    Sophie Dulauroy, Selene E Di Carlo, Gérard Eberl & Lucie Peduto

  3. Institut Pasteur, Centre d'Ingénierie Génétique Murine, Paris, France

    Francina Langa

Authors
  1. Sophie Dulauroy
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Selene E Di Carlo
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Francina Langa
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Gérard Eberl
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Lucie Peduto
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

S.D. and S.E.D.C. carried out experiments and analyzed data. F.L. carried out BAC-mice transgenesis. G.E. participated in experiment design and data analysis, and wrote the manuscript. L.P. designed and directed the project, carried out experiments, analyzed data and wrote the manuscript.

Corresponding author

Correspondence to Lucie Peduto.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–8 (PDF 5580 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Dulauroy, S., Di Carlo, S., Langa, F. et al. Lineage tracing and genetic ablation of ADAM12+ perivascular cells identify a major source of profibrotic cells during acute tissue injury. Nat Med 18, 1262–1270 (2012). https://doi.org/10.1038/nm.2848

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nm.2848

This article is cited by

Search

Advanced search

Quick links

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