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EndMT contributes to the onset and progression of cerebral cavernous malformations

Nature volume 498, pages 492–496 (2013)Cite this article

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Abstract

Cerebral cavernous malformation (CCM) is a vascular dysplasia, mainly localized within the brain and affecting up to 0.5% of the human population. CCM lesions are formed by enlarged and irregular blood vessels that often result in cerebral haemorrhages. CCM is caused by loss-of-function mutations in one of three genes, namely CCM1 (also known as KRIT1), CCM2 (OSM) and CCM3 (PDCD10), and occurs in both sporadic and familial forms1. Recent studies2,3,4,5,6,7 have investigated the cause of vascular dysplasia and fragility in CCM, but the in vivo functions of this ternary complex remain unclear8. Postnatal deletion of any of the three Ccm genes in mouse endothelium results in a severe phenotype, characterized by multiple brain vascular malformations that are markedly similar to human CCM lesions9. Endothelial-to-mesenchymal transition (EndMT) has been described in different pathologies, and it is defined as the acquisition of mesenchymal- and stem-cell-like characteristics by the endothelium10,11,12. Here we show that endothelial-specific disruption of the Ccm1 gene in mice induces EndMT, which contributes to the development of vascular malformations. EndMT in CCM1-ablated endothelial cells is mediated by the upregulation of endogenous BMP6 that, in turn, activates the transforming growth factor-β (TGF-β) and bone morphogenetic protein (BMP) signalling pathway. Inhibitors of the TGF-β and BMP pathway prevent EndMT both in vitro and in vivo and reduce the number and size of vascular lesions in CCM1-deficient mice. Thus, increased TGF-β and BMP signalling, and the consequent EndMT of CCM1-null endothelial cells, are crucial events in the onset and progression of CCM disease. These studies offer novel therapeutic opportunities for this severe, and so far incurable, pathology.

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Figure 1: Ccm1 deletion causes EndMT in vivo.
Figure 2: Ccm1 deletion induces EndMT via TGF-β signalling activation.
Figure 3: TGF-β signalling inhibition reduces number and size of lesions and vessel leakage.
Figure 4: CCM1 deletion induces EndMT via BMP6 upregulation.

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References

  1. Bergametti, F. et al. Mutations within the programmed cell death 10 gene cause cerebral cavernous malformations. Am. J. Hum. Genet. 76, 42–51 (2005)

    Article  CAS  Google Scholar 

  2. Whitehead, K. J. et al. The cerebral cavernous malformation signaling pathway promotes vascular integrity via Rho GTPases. Nature Med. 15, 177–184 (2009)

    Article  CAS  Google Scholar 

  3. Chan, A. C. et al. Mutations in 2 distinct genetic pathways result in cerebral cavernous malformations in mice. J. Clin. Invest. 121, 1871–1881 (2011)

    Article  ADS  CAS  Google Scholar 

  4. Glading, A., Han, J., Stockton, R. A. & Ginsberg, M. H. KRIT-1/CCM1 is a Rap1 effector that regulates endothelial cell cell junctions. J. Cell Biol. 179, 247–254 (2007)

    Article  CAS  Google Scholar 

  5. Stockton, R. A., Shenkar, R., Awad, I. A. & Ginsberg, M. H. Cerebral cavernous malformations proteins inhibit Rho kinase to stabilize vascular integrity. J. Exp. Med. 207, 881–896 (2010)

    Article  CAS  Google Scholar 

  6. Lampugnani, M. G. et al. CCM1 regulates vascular-lumen organization by inducing endothelial polarity. J. Cell Sci. 123, 1073–1080 (2010)

    Article  CAS  Google Scholar 

  7. Wüstehube, J. et al. Cerebral cavernous malformation protein CCM1 inhibits sprouting angiogenesis by activating DELTA-NOTCH signaling. Proc. Natl Acad. Sci. USA 107, 12640–12645 (2010)

    Article  ADS  Google Scholar 

  8. Faurobert, E. & Albiges-Rizo, C. Recent insights into cerebral cavernous malformations: a complex jigsaw puzzle under construction. FEBS J. 277, 1084–1096 (2010)

    Article  CAS  Google Scholar 

  9. Boulday, G. et al. Developmental timing of CCM2 loss influences cerebral cavernous malformations in mice. J. Exp. Med. 208, 1835–1847 (2011)

    Article  CAS  Google Scholar 

  10. Zeisberg, E. M. et al. Endothelial-to-mesenchymal transition contributes to cardiac fibrosis. Nature Med. 13, 952–961 (2007)

    Article  CAS  Google Scholar 

  11. Medici, D. et al. Conversion of vascular endothelial cells into multipotent stem-like cells. Nature Med. 16, 1400–1406 (2010)

    Article  CAS  Google Scholar 

  12. Kalluri, R. & Weinberg, R. A. The basics of epithelial-mesenchymal transition. J. Clin. Invest. 119, 1420–1428 (2009)

    Article  CAS  Google Scholar 

  13. Labauge, P., Denier, C., Bergametti, F. & Tournier-Lasserve, E. Genetics of cavernous angiomas. Lancet Neurol. 6, 237–244 (2007)

    Article  CAS  Google Scholar 

  14. Lopez, D., Niu, G., Huber, P. & Carter, W. B. Tumor-induced upregulation of Twist, Snail, and Slug represses the activity of the human VE-cadherin promoter. Arch. Biochem. Biophys. 482, 77–82 (2009)

    Article  CAS  Google Scholar 

  15. Mariotti, A., Perotti, A., Sessa, C. & Ruegg, C. N-cadherin as a therapeutic target in cancer. Expert Opin. Investig. Drugs 16, 451–465 (2007)

    Article  CAS  Google Scholar 

  16. 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  Google Scholar 

  17. Li, Y., Yang, J., Luo, J. H., Dedhar, S. & Liu, Y. Tubular epithelial cell dedifferentiation is driven by the helix-loop-helix transcriptional inhibitor Id1. J. Am. Soc. Nephrol. 18, 449–460 (2007)

    Article  Google Scholar 

  18. Vivien, C. et al. Non-viral expression of mouse Oct4, Sox2, and Klf4 transcription factors efficiently reprograms tadpole muscle fibers in vivo. J. Biol. Chem. 287, 7427–7435 (2012)

    Article  CAS  Google Scholar 

  19. Gauger, K. J., Chenausky, K. L., Murray, M. E. & Schneider, S. S. SFRP1 reduction results in an increased sensitivity to TGF-β signaling. BMC Cancer 11, 59 (2011)

    Article  CAS  Google Scholar 

  20. Melisi, D. et al. LY2109761, a novel transforming growth factor β receptor type I and type II dual inhibitor, as a therapeutic approach to suppressing pancreatic cancer metastasis. Mol. Cancer Ther. 7, 829–840 (2008)

    Article  CAS  Google Scholar 

  21. Tanaka, H. et al. Transforming growth factor beta signaling inhibitor, SB-431542, induces maturation of dendritic cells and enhances anti-tumor activity. Oncol. Rep. 24, 1637–1643 (2010)

    CAS  PubMed  Google Scholar 

  22. McLean, K. et al. Human ovarian carcinoma-associated mesenchymal stem cells regulate cancer stem cells and tumorigenesis via altered BMP production. J. Clin. Invest. 121, 3206–3219 (2011)

    Article  CAS  Google Scholar 

  23. Ao, A., Hao, J., Hopkins, C. R. & Hong, C. C. DMH1, a novel BMP small molecule inhibitor, increases cardiomyocyte progenitors and promotes cardiac differentiation in mouse embryonic stem cells. PLoS ONE 7, e41627 (2012)

    Article  ADS  CAS  Google Scholar 

  24. Murtaugh, L. C., Stanger, B. Z., Kwan, K. M. & Melton, D. A. Notch signaling controls multiple steps of pancreatic differentiation. Proc. Natl Acad. Sci. USA 100, 14920–14925 (2003)

    Article  ADS  CAS  Google Scholar 

  25. Korpal, M. & Kang, Y. Targeting the transforming growth factor-beta signalling pathway in metastatic cancer. Eur. J. Cancer 46, 1232–1240 (2010)

    Article  CAS  Google Scholar 

  26. Yuan, Q. et al. Fluorofenidone suppresses epithelial-mesenchymal transition and the expression of connective tissue growth factor via inhibiting TGF-β/Smads signaling in human proximal tubular epithelial cells. Pharmazie 66, 961–967 (2011)

    CAS  PubMed  Google Scholar 

  27. Glading, A. J. & Ginsberg, M. H. Rap1 and its effector KRIT1/CCM1 regulate β-catenin signaling. Dis. Model Mech. 3, 73–83 (2010)

    Article  CAS  Google Scholar 

  28. Liebner, S. et al. β-catenin is required for endothelial-mesenchymal transformation during heart cushion development in the mouse. J. Cell Biol. 166, 359–367 (2004)

    Article  CAS  Google Scholar 

  29. Liebner, S., Czupalla, C. J. & Wolburg, H. Current concepts of blood–brain barrier development. Int. J. Dev. Biol. 55, 467–476 (2011)

    Article  CAS  Google Scholar 

  30. Louvi, A. et al. Loss of cerebral cavernous malformation 3 (Ccm3) in neuroglia leads to CCM and vascular pathology. Proc. Natl Acad. Sci. USA 108, 3737–3742 (2011)

    Article  ADS  CAS  Google Scholar 

  31. Corada, M. et al. The Wnt/β-catenin pathway modulates vascular remodeling and specification by upregulating Dll4/Notch signaling. Dev. Cell 18, 938–949 (2010)

    Article  CAS  Google Scholar 

  32. Soriano, P. Generalized lacZ expression with the ROSA26 Cre reporter strain. Nature Genet. 21, 70–71 (1999)

    Article  CAS  Google Scholar 

  33. Spagnuolo, R. et al. Gas1 is induced by VE-cadherin and vascular endothelial growth factor and inhibits endothelial cell apoptosis. Blood 103, 3005–3012 (2004)

    Article  CAS  Google Scholar 

  34. Felici, A. et al. TLP, a novel modulator of TGF-β signaling, has opposite effects on Smad2- and Smad3-dependent signaling. EMBO J. 22, 4465–4477 (2003)

    Article  CAS  Google Scholar 

  35. Bussolino, F. et al. Murine endothelioma cell lines transformed by polyoma middle T oncogene as target for and producers of cytokines. J. Immunol. 147, 2122–2129 (1991)

    CAS  PubMed  Google Scholar 

  36. Liebner, S. et al. Wnt/β-catenin signaling controls development of the blood-brain barrier. J. Cell Biol. 183, 409–417 (2008)

    Article  CAS  Google Scholar 

  37. Korff, T. & Augustin, H. G. Tensional forces in fibrillar extracellular matrices control directional capillary sprouting. J. Cell Sci. 112, 3249–3258 (1999)

    CAS  PubMed  Google Scholar 

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Acknowledgements

This work was supported by grants from: Fondation Leducq Transatlantic Network of Excellence, Associazione Italiana per la Ricerca sul Cancro (AIRC) and a ‘Special Program Molecular Clinical Oncology 5x1000’ to AIRC-Gruppo Italiano Malattie Mieloproliferative (AGIMM); the European Community: European Research Council (ERC grant 268870 call identifier ERC-2010-SdG) (EU Networks: EUSTROKE-contract-202213, OPTISTEM-contract-223098, ENDOSTEM-HEALTH-2009-241440, ENDOSTEM-HEALTH-2009-241440, JUSTBRAIN-HEALTH-2009-241861, ITN-VESSEL), and the CARIPLO Foundation. We are strongly indebted to R. Adams and D. A. Melton for sharing Cdh5(PAC)-CreERT2 and NotchIC mice, respectively. We are grateful to M. Forni and the Italian research network for Cerebral Cavernous Malformation (CCM Italia; http://www.ccmitalia.unito.it) for contributing to the study with important biological materials.

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Author notes

  1. Luigi Maddaluno and Noemi Rudini: These authors contributed equally to this work.

Authors and Affiliations

  1. IFOM Fondazione, FIRC Institute of Molecular Oncology, 20139 Milan, Italy,

    Luigi Maddaluno, Noemi Rudini, Roberto Cuttano, Luca Bravi, Costanza Giampietro, Monica Corada, Luca Ferrarini, Fabrizio Orsenigo, Eleanna Papa, Maria Grazia Lampugnani & Elisabetta Dejana

  2. INSERM, UMR-S740, F-75010 Paris, France,

    Gwenola Boulday & Elisabeth Tournier-Lasserve

  3. Université Paris Diderot, Sorbonne Paris Cité, Génétique des Maladies Vasculaires UMR-S740, F-75010 Paris, France,

    Gwenola Boulday & Elisabeth Tournier-Lasserve

  4. AP-HP, Groupe Hospitalier Saint-Louis Lariboisiere-Fernand-Widal, F-75010 Paris, France,

    Elisabeth Tournier-Lasserve

  5. Department of Pathology, Centre Hospitalier Universitaire, Avenue Côte de Nacre, 14032 Caen, France,

    Françoise Chapon

  6. IEO – European Institute of Oncology, 20139 Milan, Italy,

    Cristina Richichi

  7. CCM Italia - Department of Clinical and Biological Sciences, University of Turin, 10043 Orbassano, Turin, Italy,

    Saverio Francesco Retta

  8. Mario Negri Institute of Pharmacological Research, 20156 Milan, Italy,

    Maria Grazia Lampugnani

  9. Department of Biosciences, University of Milan, 20133 Milan, Italy,

    Elisabetta Dejana

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Contributions

Experiments were designed by L.M., N.R. and E.D.; in vivo treatments were performed by L.M., N.R., R.C., M.C. and L.B.; G.B. and E.T.-L. contributed to the scientific discussion and the setting-up of the in vivo experiments. Cell isolation and treatments were performed by L.M., N.R., E.P., M.G.L., C.G. and M.C.; qRT–PCR analyses were performed by L.M., N.R. and L.F.; F.O., R.C., C.R. and L.M. performed analysis of the retinas; human samples were analysed by L.M. with the help of F.C. and S.F.R.; the manuscript was assembled and written by L.M., N.R., C.G. and E.D.

Corresponding authors

Correspondence to Luigi Maddaluno or Elisabetta Dejana.

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The authors declare no competing financial interests.

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Maddaluno, L., Rudini, N., Cuttano, R. et al. EndMT contributes to the onset and progression of cerebral cavernous malformations. Nature 498, 492–496 (2013). https://doi.org/10.1038/nature12207

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