Most of the currently available drugs for osteoporosis inhibit osteoclastic bone resorption; only a few drugs promote osteoblastic bone formation. It is thus becoming increasingly necessary to identify the factors that regulate bone formation. We found that osteoclasts express semaphorin 4D (Sema4D), previously shown to be an axon guidance molecule, which potently inhibits bone formation. The binding of Sema4D to its receptor Plexin-B1 on osteoblasts resulted in the activation of the small GTPase RhoA, which inhibits bone formation by suppressing insulin-like growth factor-1 (IGF-1) signaling and by modulating osteoblast motility. Sema4d−/− mice, Plxnb1−/− mice and mice expressing a dominant-negative RhoA specifically in osteoblasts showed an osteosclerotic phenotype due to augmented bone formation. Notably, Sema4D-specific antibody treatment markedly prevented bone loss in a model of postmenopausal osteoporosis. Thus, Sema4D has emerged as a new therapeutic target for the discovery and development of bone-increasing drugs.
Subscribe to Journal
Get full journal access for 1 year
only $17.42 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Martin, T.J. & Sims, N.A. Osteoclast-derived activity in the coupling of bone formation to resorption. Trends Mol. Med. 11, 76–81 (2005).
Hayden, J.M., Mohan, S. & Baylink, D.J. The insulin-like growth factor system and the coupling of formation to resorption. Bone 17, 93S–98S (1995).
Hattner, R., Epker, B.N. & Frost, H.M. Suggested sequential mode of control of changes in cell behaviour in adult bone remodelling. Nature 206, 489–490 (1965).
Takahashi, H., Epker, B. & Frost, H.M. Resorption precedes formative activity. Surg. Forum 15, 437–438 (1964).
Tang, Y. et al. TGF-β1−induced migration of bone mesenchymal stem cells couples bone resorption with formation. Nat. Med. 15, 757–765 (2009).
Matsuo, K. & Irie, N. Osteoclast-osteoblast communication. Arch. Biochem. Biophys. 473, 201–209 (2008).
Parfitt, A.M., Mundy, G.R., Roodman, G.D., Hughes, D.E. & Boyce, B.F. A new model for the regulation of bone resorption, with particular reference to the effects of bisphosphonates. J. Bone Miner. Res. 11, 150–159 (1996).
Larrivée, B., Freitas, C., Suchting, S., Brunet, I. & Eichmann, A. Guidance of vascular development: lessons from the nervous system. Circ. Res. 104, 428–441 (2009).
Suzuki, K., Kumanogoh, A. & Kikutani, H. Semaphorins and their receptors in immune cell interactions. Nat. Immunol. 9, 17–23 (2008).
Tran, T.S., Kolodkin, A.L. & Bharadwaj, R. Semaphorin regulation of cellular morphology. Annu. Rev. Cell Dev. Biol. 23, 263–292 (2007).
Huber, A.B., Kolodkin, A.L., Ginty, D.D. & Cloutier, J.F. Signaling at the growth cone: ligand-receptor complexes and the control of axon growth and guidance. Annu. Rev. Neurosci. 26, 509–563 (2003).
Dickson, B.J. Molecular mechanisms of axon guidance. Science 298, 1959–1964 (2002).
Tamagnone, L. & Comoglio, P.M. Signalling by semaphorin receptors: cell guidance and beyond. Trends Cell Biol. 10, 377–383 (2000).
Irie, N. et al. Bidirectional signaling through ephrinA2-EphA2 enhances osteoclastogenesis and suppresses osteoblastogenesis. J. Biol. Chem. 284, 14637–14644 (2009).
Sutton, A.L. et al. Semaphorin 3B is a 1,25-dihydroxyvitamin D3-induced gene in osteoblasts that promotes osteoclastogenesis and induces osteopenia in mice. Mol. Endocrinol. 22, 1370–1381 (2008).
Takegahara, N. et al. Plexin-A1 and its interaction with DAP12 in immune responses and bone homeostasis. Nat. Cell Biol. 8, 615–622 (2006).
Zhao, C. et al. Bidirectional ephrinB2-EphB4 signaling controls bone homeostasis. Cell Metab. 4, 111–121 (2006).
Delorme, G., Saltel, F., Bonnelye, E., Jurdic, P. & Machuca-Gayet, I. Expression and function of semaphorin 7A in bone cells. Biol. Cell 97, 589–597 (2005).
Kumanogoh, A. et al. Identification of CD72 as a lymphocyte receptor for the class IV semaphorin CD100: a novel mechanism for regulating B cell signaling. Immunity 13, 621–631 (2000).
Elhabazi, A., Delaire, S., Bensussan, A., Boumsell, L. & Bismuth, G. Biological activity of soluble CD100. I. The extracellular region of CD100 is released from the surface of T lymphocytes by regulated proteolysis. J. Immunol. 166, 4341–4347 (2001).
Tamagnone, L. et al. Plexins are a large family of receptors for transmembrane, secreted, and GPI-anchored semaphorins in vertebrates. Cell 99, 71–80 (1999).
Swiercz, J.M., Worzfeld, T. & Offermanns, S. ErbB-2 and Met reciprocally regulate cellular signaling via plexin-B1. J. Biol. Chem. 283, 1893–1901 (2008).
Kruger, R.P., Aurandt, J. & Guan, K.L. Semaphorins command cells to move. Nat. Rev. Mol. Cell Biol. 6, 789–800 (2005).
Oinuma, I., Ishikawa, Y., Katoh, H. & Negishi, M. The Semaphorin 4D receptor Plexin-B1 is a GTPase activating protein for R-Ras. Science 305, 862–865 (2004).
Driessens, M.H., Olivo, C., Nagata, K., Inagaki, M. & Collard, J.G. B plexins activate Rho through PDZ-RhoGEF. FEBS Lett. 529, 168–172 (2002).
Perrot, V., Vazquez-Prado, J. & Gutkind, J.S. Plexin B regulates Rho through the guanine nucleotide exchange factors leukemia-associated Rho GEF (LARG) and PDZ-RhoGEF. J. Biol. Chem. 277, 43115–43120 (2002).
Swiercz, J.M., Kuner, R., Behrens, J. & Offermanns, S. Plexin-B1 directly interacts with PDZ-RhoGEF/LARG to regulate RhoA and growth cone morphology. Neuron 35, 51–63 (2002).
Kobayashi, K. et al. Survival of developing motor neurons mediated by Rho GTPase signaling pathway through Rho-kinase. J. Neurosci. 24, 3480–3488 (2004).
Dacquin, R., Starbuck, M., Schinke, T. & Karsenty, G. Mouse α1(I)-collagen promoter is the best known promoter to drive efficient Cre recombinase expression in osteoblast. Dev. Dyn. 224, 245–251 (2002).
Sordella, R. et al. Modulation of CREB activity by the Rho GTPase regulates cell and organism size during mouse embryonic development. Dev. Cell 2, 553–565 (2002).
Gomez, C. et al. Expression of Semaphorin-3A and its receptors in endochondral ossification: potential role in skeletal development and innervation. Dev. Dyn. 234, 393–403 (2005).
Soker, S., Takashima, S., Miao, H.Q., Neufeld, G. & Klagsbrun, M. Neuropilin-1 is expressed by endothelial and tumor cells as an isoform-specific receptor for vascular endothelial growth factor. Cell 92, 735–745 (1998).
Behar, O., Golden, J.A., Mashimo, H., Schoen, F.J. & Fishman, M.C. Semaphorin III is needed for normal patterning and growth of nerves, bones and heart. Nature 383, 525–528 (1996).
Sekido, Y. et al. Human semaphorins A(V) and IV reside in the 3p21.3 small cell lung cancer deletion region and demonstrate distinct expression patterns. Proc. Natl. Acad. Sci. USA 93, 4120–4125 (1996).
Kitsukawa, T., Shimono, A., Kawakami, A., Kondoh, H. & Fujisawa, H. Overexpression of a membrane protein, neuropilin, in chimeric mice causes anomalies in the cardiovascular system, nervous system and limbs. Development 121, 4309–4318 (1995).
Koh, J.M. et al. Association study of semaphorin 7a (sema7a) polymorphisms with bone mineral density and fracture risk in postmenopausal Korean women. J. Hum. Genet. 51, 112–117 (2006).
Kumanogoh, A. et al. Requirement for the lymphocyte semaphorin, CD100, in the induction of antigen-specific T cells and the maturation of dendritic cells. J. Immunol. 169, 1175–1181 (2002).
Delaire, S. et al. Biological activity of soluble CD100. II. Soluble CD100, similarly to H-SemaIII, inhibits immune cell migration. J. Immunol. 166, 4348–4354 (2001).
Hirschberg, A. et al. Gene deletion mutants reveal a role for semaphorin receptors of the plexin-B family in mechanisms underlying corticogenesis. Mol. Cell. Biol. 30, 764–780 (2010).
Di Benedetto, A. et al. N-cadherin and cadherin 11 modulate postnatal bone growth and osteoblast differentiation by distinct mechanisms. J. Cell Sci. 123, 2640–2648 (2010).
Diarra, D. et al. Dickkopf-1 is a master regulator of joint remodeling. Nat. Med. 13, 156–163 (2007).
Matsumoto, T. & Abe, M. TGF-β-related mechanisms of bone destruction in multiple myeloma. Bone 48, 129–134 (2011).
Yadav, V.K. et al. Pharmacological inhibition of gut-derived serotonin synthesis is a potential bone anabolic treatment for osteoporosis. Nat. Med. 16, 308–312 (2010).
Shi, W. et al. The class IV semaphorin CD100 plays nonredundant roles in the immune system: defective B and T cell activation in CD100-deficient mice. Immunity 13, 633–642 (2000).
Friedel, R.H. et al. Gene targeting using a promoterless gene trap vector (“targeted trapping”) is an efficient method to mutate a large fraction of genes. Proc. Natl. Acad. Sci. USA 102, 13188–13193 (2005).
Nishikawa, K. et al. Maf promotes osteoblast differentiation in mice by mediating the age-related switch in mesenchymal cell differentiation. J. Clin. Invest. 120, 3455–3465 (2010).
Kim, S. et al. Stat1 functions as a cytoplasmic attenuator of Runx2 in the transcriptional program of osteoblast differentiation. Genes Dev. 17, 1979–1991 (2003).
Takayanagi, H. et al. Induction and activation of the transcription factor NFATc1 (NFAT2) integrate RANKL signaling in terminal differentiation of osteoclasts. Dev. Cell 3, 889–901 (2002).
Koga, T. et al. NFAT and Osterix cooperatively regulate bone formation. Nat. Med. 11, 880–885 (2005).
Gregory, C.A., Gunn, W.G., Peister, A. & Prockop, D.J. An Alizarin red-based assay of mineralization by adherent cells in culture: comparison with cetylpyridinium chloride extraction. Anal. Biochem. 329, 77–84 (2004).
Okuno, T. et al. Roles of Sema4D-plexin-B1 interactions in the central nervous system for pathogenesis of experimental autoimmune encephalomyelitis. J. Immunol. 184, 1499–1506 (2010).
Regev, A., Goldman, S. & Shalev, E. Expression of plexin-B1 in the mouse ovary and its possible role in follicular development. Fertil. Steril. 84 (suppl 2), 1210–1219 (2005).
Shinohara, M. et al. Nox1 redox signaling mediates oncogenic Ras-induced disruption of stress fibers and focal adhesions by down-regulating Rho. J. Biol. Chem. 282, 17640–17648 (2007).
Shinohara, M. et al. Tyrosine kinases Btk and Tec regulate osteoclast differentiation by linking RANK and ITAM signals. Cell 132, 794–806 (2008).
Sotobori, T. et al. Bone morphogenetic protein-2 promotes the haptotactic migration of murine osteoblastic and osteosarcoma cells by enhancing incorporation of integrin 1 into lipid rafts. Exp. Cell Res. 312, 3927–3938 (2006).
We are grateful to G. Karsenty (Columbia University), K. Kobayashi (Fukushima Medical University School of Medicine), A. Kumanogoh (Osaka University) and T. Kitamura (The University of Tokyo) for kindly providing α(1)I-Cre mice, CAT-RhoA DN mice, recombinant Fc-sema4D protein, and the retrovirus vectors and Plat-E cells, respectively. We also thank E. Sumiya, A. Suematsu, Y. Kunisawa, T. Ando, K. Okamoto, T. Nakashima, M. Oh-hora, M. Hayashi, A. Terashima, Y. Nagai and H. Negishi for discussion and assistance. This work was supported in part by a grant for the Exploratory Research for Advanced Technology, Takayanagi Osteonetwork Project from the Japan Science and Technology Agency, Grant-in-Aid for Young Scientist A, Grant-in-Aid for Challenging Exploratory Research and Grant-in-Aid for JSPS Fellows from the Japan Society for the Promotion of Science, a grant for the Global Center of Excellence Program from the Ministry of Education, Culture, Sports, Science and Technology of Japan, and grants from the Ichiro Kanehara Foundation, the Uehara Memorial Foundation and the Naito Foundation.
The authors declare no competing financial interests.
About this article
Cite this article
Negishi-Koga, T., Shinohara, M., Komatsu, N. et al. Suppression of bone formation by osteoclastic expression of semaphorin 4D. Nat Med 17, 1473–1480 (2011). https://doi.org/10.1038/nm.2489
Journal of Bone and Mineral Metabolism (2021)
Pharmacological Research (2020)