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Brain-type creatine kinase has a crucial role in osteoclast-mediated bone resorption

Nature Medicine volume 14, pages 966972 (2008) | Download Citation

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Abstract

Osteoclasts differentiate from precursor cells of the monocyte-macrophage lineage and subsequently become activated to be competent for bone resorption through programs primarily governed by receptor activator of nuclear factor-κB ligand in cooperation with macrophage colony–stimulating factor1,2,3. Proteins prominently expressed at late phases of osteoclastogenesis and with a supportive role in osteoclast function are potential therapeutic targets for bone-remodeling disorders. In this study, we used a proteomics approach to show that abundance of the brain-type cytoplasmic creatine kinase (Ckb) is greatly increased during osteoclastogenesis. Decreasing Ckb abundance by RNA interference or blocking its enzymatic activity with a pharmacological inhibitor, cyclocreatine, suppressed the bone-resorbing activity of osteoclasts grown in vitro via combined effects on actin ring formation, RhoA GTPase activity and vacuolar ATPase function. Activities of osteoclasts derived from Ckb−/− mice were similarly affected. In vivo studies showed that Ckb−/− mice were better protected against bone loss induced by ovariectomy, lipopolysaccharide challenge or interleukin-1 treatment than wild-type controls. Furthermore, administration of cyclocreatine or adenoviruses harboring Ckb small hairpin RNA attenuated bone loss in rat and mouse models. Our findings establish an important role for Ckb in the bone-resorbing function of osteoclasts and underscore its potential as a new molecular target for antiresorptive drug development.

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Acknowledgements

We thank S.I. Kim, H.B. Kwak, J.Y. Yang, T.K. Yu and J.S. Ko for technical help and discussion. We also thank T. Kitamura (University of Tokyo) for Plat-E cells. This work was supported by the 21C Frontier Functional Proteomics Project grants FPR08B1-170 (to H.-H.K.) and FPR08A1-070 (to Y.K.P.), the Research Program for New Drug Target Discovery grant M10748000257-07N4800-25710 (to H.-H.K.) from the Ministry of Education, Science & Technology, Korea and NKB-KWF grant 2002-2763 (to B.W.).

Author information

Affiliations

  1. Department of Cell and Developmental Biology, Brain Korea 21, School of Dentistry, Seoul National University, 28 Yeongon-Dong, Jongno-gu, Seoul 110-749, Korea.

    • Eun-Ju Chang
    • , Jeongim Ha
    • , Jiyoon Ryu
    • , Hyung Joon Kim
    • , Youngkyun Lee
    • , Hyun-Man Kim
    • , Zang Hee Lee
    •  & Hong-Hee Kim
  2. Department of Cell Biology, Nijmegen Centre for Molecular Life Sciences, Radboud University Medical Centre, P.O. Box 9101, 6500 HB Nijmegen, The Netherlands.

    • Frank Oerlemans
    •  & Bé Wieringa
  3. Research Division, Jeonnam Biotechnology Research Center, 121 Naepyeongri, Hwasun-gun, Jeollanamdo 519-801, Korea.

    • You Jin Lee
  4. Department of Microbiology, Center for Viral Disease Research, College of Medicine, Inje University, 633-165 Gaegeum-Dong, Busanjin-gu, Busan 614-735, Korea.

    • Soo Woong Lee
  5. Department of Biochemistry and Cell Biology, School of Medicine and Skeletal Diseases Genome Research Center, Kyungpook National University, 101 Dong-in Dong, Jung-gu, Daegu 700-422, Korea.

    • Je-Yong Choi
  6. Korea Basic Science Institute, 52 Eoeun-Dong, Yusung-gu, Daejeon 305-333, Korea.

    • Jin Young Kim
  7. Department of Internal Medicine, Seoul National University College of Medicine, 28 Yeongon-Dong, Jongno-gu, Seoul 110-744, Korea.

    • Chan Soo Shin
  8. Age-related and Brain Disease Research Center, Department of Nanopharmaceutical and Life Sciences, Kyung Hee University, 1 Hoegi-Dong, Dongdaemun-gu, Seoul 130-701, Korea.

    • Youngmi Kim Pak
  9. Department of Orthopaedic Surgery, Faculty of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-gu, Tokyo 113-0033, Japan.

    • Sakae Tanaka

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Contributions

E.-J.C. designed and performed most of the experiments and wrote the manuscript. J.H. performed the experiments by assisting with tissue collection; DNA, RNA and protein isolation; and analyses. F.O. performed the experiments by assisting with breeding and analyses of Ckb-knockout mice. Y.J.L. carried out histological analysis. J.R. and H.J.K. performed the V-ATPase assay. Y.L., H.-M.K. and J.-Y.C. participated in histomorphometrical analysis. S.W.L and J.Y.K. performed proteomic experiments. Y.K.P. generated adenoviruses. C.S.S., Z.H.L. and S.T. participated in generation and analyses of various in vivo bone resorption models. B.W. was responsible for establishing Ckb-knockout mice and participated in data interpretation and discussion. Z.H.L. and H.-H.K. conceived the study. H.-H.K. supervised the study and wrote the manuscript.

Corresponding authors

Correspondence to Bé Wieringa or Hong-Hee Kim.

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DOI

https://doi.org/10.1038/nm.1860

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