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Costimulatory signals mediated by the ITAM motif cooperate with RANKL for bone homeostasis

Nature volume 428, pages 758–763 (2004)Cite this article

Abstract

Costimulatory signals are required for activation of immune cells1, but it is not known whether they contribute to other biological systems. The development and homeostasis of the skeletal system depend on the balance between bone formation and resorption2,3. Receptor activator of NF-κB ligand (RANKL) regulates the differentiation of bone-resorbing cells, osteoclasts, in the presence of macrophage-colony stimulating factor (M-CSF)4,5. But it remains unclear how RANKL activates the calcium signals that lead to induction of nuclear factor of activated T cells c1, a key transcription factor for osteoclastogenesis6. Here we show that mice lacking immunoreceptor tyrosine-based activation motif (ITAM)7-harbouring adaptors8,9,10, Fc receptor common γ subunit (FcRγ) and DNAX-activating protein (DAP)12, exhibit severe osteopetrosis owing to impaired osteoclast differentiation. In osteoclast precursor cells, FcRγ and DAP12 associate with multiple immunoreceptors11,12,13,14,15 and activate calcium signalling through phospholipase Cγ. Thus, ITAM-dependent costimulatory signals activated by multiple immunoreceptors are essential for the maintenance of bone homeostasis. These results reveal that RANKL and M-CSF are not sufficient to activate the signals required for osteoclastogenesis.

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Figure 1: Impaired osteoclastogenesis in the absence of DAP12 and the compensatory mechanism by osteoblasts.
Figure 2: Severe osteopetrosis in DAP12-/- FcRγ-/- (DKO) mice due to impaired osteoclast differentiation.
Figure 3: Distinct roles of FcRγ- and DAP12-associating immunoreceptors in the regulation of osteoclastogenesis.
Figure 4: ITAM-harbouring adaptors are essential for RANKL induction of calcium signalling and NFATc1.

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Acknowledgements

We thank J. V. Ravetch, H. Kubagawa and M. D. Cooper for providing materials, and M. Kaji, A. Sugahara, Y. Ito, K. Maya, A. Sato, A. Nakamura, M. Isobe, T. Yokochi, A. Izumi, T. Kohro, Y. Matsui, H. Murayama, K. Sato, M. Asagiri and I. Kawai for technical assistance and discussion. This work was supported in part by a grant for Advanced Research on Cancer from the Ministry of Education, Culture, Sports, Science, and Technology of Japan, the CREST and PRESTO programs of Japan Science and Technology Agency (JST), grants for the 21st century COE program ‘Frontier Research on Molecular Destruction and Reconstruction of Tooth and Bone’ and ‘Center for Innovative Therapeutic Development Towards the Conquest of Signal Transduction Diseases’, Grants-in-Aid for Scientific Research from JSPS and MEXT, Health Sciences Research Grants from the Ministry of Health, Labour and Welfare of Japan, grants of the Virtual Research Institute of Aging of Nippon Boehringer Ingelheim, Mochida Medical and Pharmaceutical Research Foundation and a grant from Japan Orthopaedics and Traumatology Foundation.

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  1. Takako Koga and Masanori Inui: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Immunology, Graduate School of Medicine and Faculty of Medicine, University of Tokyo, Hongo 7-3-1, Bunkyo-ku, 113-0033, Tokyo, Japan

    Takako Koga, Sunhwa Kim, Ayako Suematsu, Tadatsugu Taniguchi & Hiroshi Takayanagi

  2. Department of Cellular Physiological Chemistry, Graduate School, Tokyo Medical and Dental University, and COE Program for Frontier Research on Molecular Destruction and Reconstruction of Tooth and Bone, Yushima 1-5-45, Bunkyo-ku, Tokyo, 113-8549, Japan

    Takako Koga, Ayako Suematsu & Hiroshi Takayanagi

  3. PRESTO, Japan Science and Technology Agency (JST), Honcho 4-1-8, Kawaguchi, 332-0012, Saitama, Japan

    Takako Koga, Ayako Suematsu & Hiroshi Takayanagi

  4. Department of Experimental Immunology, Institute of Development, Aging and Cancer, Tohoku University, Seiryo 4-1, Aoba-ku, Sendai, 980-8575, Japan

    Masanori Inui, Kazuya Inoue, Eiji Kobayashi, Toshio Iwata & Toshiyuki Takai

  5. CREST, JST, Honcho 4-1-8, Kawaguchi, 332-0012, Saitama, Japan

    Masanori Inui, Kazuya Inoue, Eiji Kobayashi, Toshio Iwata & Toshiyuki Takai

  6. Biosignal Research Center, Institute for Molecular and Cellular Regulation, Gunma University, Showa-machi 3-39-15, 371-8512, Maebashi, Japan

    Hiroshi Ohnishi & Takashi Matozaki

  7. Department of Molecular Biology and Medicine, Research Center for Advanced Science and Technology, University of Tokyo, Komaba 4-6-1, Meguro-ku, 153-8904, Tokyo, Japan

    Tatsuhiko Kodama

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  1. Takako Koga
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Correspondence to Hiroshi Takayanagi or Toshiyuki Takai.

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Supplementary information

Supplementary Figures

Supplementary figure 1: Analysis of the rescue effects by osteoblast and the involvement of FcRγ; Supplementary figure 2: Bone morphometric and histological analysis of DAP12-/-FcRg-/- (DKO) mice; Supplementary figure 3: ITAM-harbouring adaptor-associated immunoreceptors in osteoclast lineage cells; Supplementary figure 4: Intracellular signalling events downstream of ITAM-harbouring adaptors. (PDF 328 kb)

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Koga, T., Inui, M., Inoue, K. et al. Costimulatory signals mediated by the ITAM motif cooperate with RANKL for bone homeostasis. Nature 428, 758–763 (2004). https://doi.org/10.1038/nature02444

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