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BMP type I receptor inhibition reduces heterotopic ossification

An Erratum to this article was published on 01 January 2009

This article has been updated

Abstract

Fibrodysplasia ossificans progressiva (FOP) is a congenital disorder of progressive and widespread postnatal ossification of soft tissues1,2,3,4 and is without known effective treatments. Affected individuals harbor conserved mutations in the ACVR1 gene that are thought to cause constitutive activation of the bone morphogenetic protein (BMP) type I receptor, activin receptor-like kinase-2 (ALK2)5. Here we show that intramuscular expression in the mouse of an inducible transgene encoding constitutively active ALK2 (caALK2), resulting from a glutamine to aspartic acid change at amino acid position 207, leads to ectopic endochondral bone formation, joint fusion and functional impairment, thus phenocopying key aspects of human FOP. A selective inhibitor of BMP type I receptor kinases, LDN-193189 (ref. 6), inhibits activation of the BMP signaling effectors SMAD1, SMAD5 and SMAD8 in tissues expressing caALK2 induced by adenovirus specifying Cre (Ad.Cre). This treatment resulted in a reduction in ectopic ossification and functional impairment. In contrast to localized induction of caALK2 by Ad.Cre (which entails inflammation), global postnatal expression of caALK2 (induced without the use of Ad.Cre and thus without inflammation) does not lead to ectopic ossification. However, if in this context an inflammatory stimulus was provided with a control adenovirus, ectopic bone formation was induced. Like LDN-193189, corticosteroid inhibits ossification in Ad.Cre-injected mutant mice, suggesting caALK2 expression and an inflammatory milieu are both required for the development of ectopic ossification in this model. These results support the role of dysregulated ALK2 kinase activity in the pathogenesis of FOP and suggest that small molecule inhibition of BMP type I receptor activity may be useful in treating FOP and heterotopic ossification syndromes associated with excessive BMP signaling.

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Figure 1: Mouse model of FOP.
Figure 2: Effect of LDN-193189 on BMP signaling and function.
Figure 3: Impact of LDN-193189 on ectopic ossification in vivo.
Figure 4: Impact of pharmacologic inhibition of BMP signaling and inflammation in the mouse FOP model.

Change history

  • 04 December 2008

    In the version of this article initially published, the title included a misspelling—‘heterotropic’ should have been ‘heterotopic’. Additionally, the fourth and fifth sentences of the abstract were incorrectly worded and have been corrected to state more clearly the role of Ad.Cre. These changes do not affect the scientific content of the text. The errors have been corrected in the HTML and PDF versions of the article.

References

  1. Shore, E.M. & Kaplan, F.S. Insights from a rare genetic disorder of extra-skeletal bone formation, fibrodysplasia ossificans progressiva (FOP). Bone 43, 427–433 (2008).

    Article  CAS  Google Scholar 

  2. Buyse, G., Silberstein, J., Goemans, N. & Casaer, P. Fibrodysplasia ossificans progressiva: still turning into wood after 300 years? Eur. J. Pediatr. 154, 694–699 (1995).

    Article  CAS  Google Scholar 

  3. Kaplan, F.S., Glaser, D.L., Pignolo, R.J. & Shore, E.M. A new era for fibrodysplasia ossificans progressiva: a druggable target for the second skeleton. Expert Opin. Biol. Ther. 7, 705–712 (2007).

    Article  CAS  Google Scholar 

  4. Kaplan, F.S. et al. Fibrodysplasia ossificans progressiva. Best Pract. Res. Clin. Rheumatol. 22, 191–205 (2008).

    Article  CAS  Google Scholar 

  5. Shore, E.M. et al. A recurrent mutation in the BMP type I receptor ACVR1 causes inherited and sporadic fibrodysplasia ossificans progressiva. Nat. Genet. 38, 525–527 (2006).

    Article  CAS  Google Scholar 

  6. Cuny, G.D. et al. Structure-activity relationship study of bone morphogenetic protein (BMP) signaling inhibitors. Bioorg. Med. Chem. Lett. 18, 4388–4392 (2008).

    Article  CAS  Google Scholar 

  7. Tsuchida, K., Mathews, L.S. & Vale, W.W. Cloning and characterization of a transmembrane serine kinase that acts as an activin type I receptor. Proc. Natl. Acad. Sci. USA 90, 11242–11246 (1993).

    Article  CAS  Google Scholar 

  8. Miyazono, K., Maeda, S. & Imamura, T. BMP receptor signaling: Transcriptional targets, regulation of signals, and signaling cross-talk. Cytokine Growth Factor Rev. 16, 251–263 (2005).

    Article  CAS  Google Scholar 

  9. Groppe, J.C., Shore, E.M. & Kaplan, F.S. Functional modeling of the ACVR1 (R206H) mutation in FOP. Clin. Orthop. Relat. Res. 462, 87–92 (2007).

    Article  Google Scholar 

  10. Shore, E.M., Xu, M., Connor, J.M. & Kaplan, F.S. Mutations in the BMP type I receptor ACVR1 in patients with fibrodysplasia ossificans progressiva (FOP). J. Bone Miner. Res. 21, S75 (2006).

    Article  Google Scholar 

  11. Macias-Silva, M., Hoodless, P.A., Tang, S.J., Buchwald, M. & Wrana, J.L. Specific activation of Smad1 signaling pathways by the BMP7 type I receptor, ALK2. J. Biol. Chem. 273, 25628–25636 (1998).

    Article  CAS  Google Scholar 

  12. Zhang, D. et al. ALK2 functions as a BMP type I receptor and induces Indian hedgehog in chondrocytes during skeletal development. J. Bone Miner. Res. 18, 1593–1604 (2003).

    Article  CAS  Google Scholar 

  13. Koefoed, M. et al. Biological effects of rAAV-caAlk2 coating on structural allograft healing. Mol. Ther. 12, 212–218 (2005).

    Article  CAS  Google Scholar 

  14. Fukuda, T. et al. Generation of a mouse with conditionally activated signaling through the BMP receptor, ALK2. Genesis 44, 159–167 (2006).

    Article  CAS  Google Scholar 

  15. Waheed, I. et al. Factors associated with induced chronic inflammation in mdx skeletal muscle cause posttranslational stabilization and augmentation of extrasynaptic sarcolemmal utrophin. Hum. Gene Ther. 16, 489–501 (2005).

    Article  CAS  Google Scholar 

  16. Yu, P.B. et al. Dorsomorphin inhibits BMP signals required for embryogenesis and iron metabolism. Nat. Chem. Biol. 4, 33–41 (2008).

    Article  CAS  Google Scholar 

  17. Yu, P.B. et al. Bone morphogenetic protein (BMP) type II receptor is required for BMP-mediated growth arrest and differentiation in pulmonary artery smooth muscle cells. J. Biol. Chem. 283, 3877–3888 (2008).

    Article  CAS  Google Scholar 

  18. Billings, P.C. et al. Dysregulated BMP signaling and enhanced osteogenic differentiation of connective tissue progenitor cells from patients with fibrodysplasia ossificans progressiva (FOP). J. Bone Miner. Res. 23, 305–313 (2008).

    Article  CAS  Google Scholar 

  19. Fukuda, T. et al. Constitutively activated ALK2 and increased smad1/5 cooperatively induce BMP signaling in fibrodysplasia ossificans progressiva. J. Biol. Chem. published online, doi: 10.1074/jbc.M801681200 (6 August 2008).

  20. Hegyi, L. et al. Stromal cells of fibrodysplasia ossificans progressiva lesions express smooth muscle lineage markers and the osteogenic transcription factor Runx2/Cbfa-1: clues to a vascular origin of heterotopic ossification? J. Pathol. 201, 141–148 (2003).

    Article  CAS  Google Scholar 

  21. Hayashi, S. & McMahon, A.P. Efficient recombination in diverse tissues by a tamoxifen-inducible form of Cre: a tool for temporally regulated gene activation/inactivation in the mouse. Dev. Biol. 244, 305–318 (2002).

    Article  CAS  Google Scholar 

  22. Scarlett, R.F. et al. Influenza-like viral illnesses and flare-ups of fibrodysplasia ossificans progressiva. Clin. Orthop. Relat. Res. 275–279 (2004).

    Article  Google Scholar 

  23. Kaplan, F.S. et al. Hematopoietic stem-cell contribution to ectopic skeletogenesis. J. Bone Joint Surg. Am. 89, 347–357 (2007).

    Article  Google Scholar 

  24. Pignolo, R.J., Suda, R.K. & Kaplan, F.S. The fibrodysplasia ossificans progressiva lesion. Clin. Rev. Bone Miner. Metab. 5, 195–200 (2005).

    Article  Google Scholar 

  25. Zhao, G.Q. Consequences of knocking out BMP signaling in the mouse. Genesis 35, 43–56 (2003).

    Article  CAS  Google Scholar 

  26. Tsuji, K. et al. BMP2 activity, although dispensable for bone formation, is required for the initiation of fracture healing. Nat. Genet. 38, 1424–1429 (2006).

    Article  CAS  Google Scholar 

  27. Corriere, M.A. et al. Endothelial Bmp4 is induced during arterial remodeling: effects on smooth muscle cell migration and proliferation. J. Surg. Res. 145, 142–149 (2008).

    Article  CAS  Google Scholar 

  28. Yu, P.B., Beppu, H., Kawai, N., Li, E. & Bloch, K.D. Bone morphogenetic protein (BMP) type II receptor deletion reveals BMP ligand–specific gain of signaling in pulmonary artery smooth muscle cells. J. Biol. Chem. 280, 24443–24450 (2005).

    Article  CAS  Google Scholar 

  29. Komori, T. et al. Targeted disruption of Cbfa1 results in a complete lack of bone formation owing to maturational arrest of osteoblasts. Cell 89, 755–764 (1997).

    Article  CAS  Google Scholar 

  30. Korchynskyi, O. & ten Dijke, P. Identification and functional characterization of distinct critically important bone morphogenetic protein–specific response elements in the Id1 promoter. J. Biol. Chem. 277, 4883–4891 (2002).

    Article  CAS  Google Scholar 

  31. Fujii, M. et al. Roles of bone morphogenetic protein type I receptors and Smad proteins in osteoblast and chondroblast differentiation. Mol. Biol. Cell 10, 3801–3813 (1999).

    Article  CAS  Google Scholar 

  32. Dennler, S. et al. Direct binding of Smad3 and Smad4 to critical TGF β–inducible elements in the promoter of human plasminogen activator inhibitor-type 1 gene. EMBO J. 17, 3091–3100 (1998).

    Article  CAS  Google Scholar 

  33. Shimizu, A. et al. Identification of receptors and Smad proteins involved in activin signalling in a human epidermal keratinocyte cell line. Genes Cells 3, 125–134 (1998).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank P. ten Dijke (Leiden University Medical Center) for providing BRE-Luc and CAGA-Luc and K. Miyazono (University of Tokyo) for providing caALK2, caALK3, caALK4, caALK5, caALK6 and caALK7. We are grateful to H. Beppu, E. Schipani, H. Kronenberg, A. Wagers, J. Groppe, W. Zapol and F. Kaplan for insightful discussions and technical expertise, A. Graveline and D. Panus for technical assistance and E. Buys for technical expertise. This work was supported by US National Institutes of Health grants HL079943 (P.B.Y.) and HL074352 (K.D.B.), the US National Institute of Environmental Health Sciences Intramural Research Program grant ES071003-10 (Y.M.) and Partners Healthcare. This work was also supported by a Howard Hughes Medical Institute Early Career Award (P.B.Y.), a Pulmonary Hypertension Association Mentored Clinical Scientist Award (P.B.Y.), a grant from the GlaxoSmithKline Research & Education Foundation for Cardiovascular Disease (P.B.Y.) and a Developmental Grant from the Center for Research in Fibrodysplasia Ossificans Progressiva and Related Disorders at the University of Pennsylvania (C.C.H.).

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Authors and Affiliations

Authors

Contributions

P.B.Y. wrote the manuscript. P.B.Y., D.Y.D. and C.S.L. designed and performed experiments and analyzed data. G.D.C., P.B.Y., K.D.B. and R.T.P. helped to design, synthesize and evaluate dorsomorphin derivatives, and G.D.C. provided pharmacokinetic data. M.L.B. provided technical expertise and μCT tomography data. D.W.H. and P.M.M. performed experiments. C.S. tested the efficacy of the dorsomorphin derivative with additional assays. N.K. performed control experiments. Y.M. and T.F. provided key experimental reagents. T.K. provided reagents and experimental advice. C.C.H., Y.M. and K.D.B. provided feedback and experimental advice, and P.B.Y. and K.D.B. edited the manuscript.

Supplementary information

Supplementary Text and Figures

Supplementary Figs. 1–7 (PDF 603 kb)

Supplementary Video 1

A wild-type mouse injected with Ad.Cre in the left hindlimb (P7), shown at P60. No deficits in limb function or gait are apparent. (MOV 1633 kb)

Supplementary Video 2

A conditional caALK2 mouse injected with Ad.Cre in the left hindlimb (P7), shown at P60. Severe fixed extension of the left hindlimb prevents use of the limb during ambulation. (MOV 3987 kb)

Supplementary Video 3

Vehicle-treated, Ad.Cre-injected conditional caALK2 mouse shown at P15. Mild impairment of gait due to decreased range-of-motion of the knee and hip joints. (MOV 2955 kb)

Supplementary Video 4

Vehicle-treated, Ad.Cre-injected conditional caALK2 mouse shown at P30. Severe gait impairment due to fixed extension of knee, hip and ankle joints. (MOV 1338 kb)

Supplementary Video 5

LDN-193189-treated, Ad.Cre-injected conditional caALK2 mouse shown at P15. No impairment of gait or range-of-motion. (MOV 2631 kb)

Supplementary Video 6

LDN-193189-treated, Ad.Cre-injected conditional caALK2 mouse shown at P30. Mild gait abnormality due to moderately decreased range of motion in knee and hip joints. (MOV 3027 kb)

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Yu, P., Deng, D., Lai, C. et al. BMP type I receptor inhibition reduces heterotopic ossification. Nat Med 14, 1363–1369 (2008). https://doi.org/10.1038/nm.1888

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