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.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Tanaka, S., Nakamura, K., Takahasi, N. & Suda, T. Role of RANKL in physiological and pathological bone resorption and therapeutics targeting the RANKL-RANK signaling system. Immunol. Rev. 208, 30–49 (2005).
Teitelbaum, S.L. Bone resorption by osteoclasts. Science 289, 1504–1508 (2000).
Boyle, W.J., Simonet, W.S. & Lacey, D.L. Osteoclast differentiation and activation. Nature 423, 337–342 (2003).
Vaananen, K. Osteoclast function: biology and mechanisms. in Principles of Bone Biology (eds. Bilezikian J.P., Raisz, L.G. & Rodan, G.A.) 103–113 (Academic Press, San Diego, 1996).
Blair, H.C., Teitelbaum, S.L., Ghiselli, R. & Gluck, S. Osteoclastic bone resorption by a polarized vacuolar proton pump. Science 245, 855–857 (1989).
Li, Y.P., Chen, W., Liang, Y., Li, E. & Stashenko, P. Atp6i-deficient mice exhibit severe osteopetrosis due to loss of osteoclast-mediated extracellular acidification. Nat. Genet. 23, 447–451 (1999).
Frattini, A. et al. Defects in TCIRG1 subunit of the vacuolar proton pump are responsible for a subset of human autosomal recessive osteopetrosis. Nat. Genet. 25, 343–346 (2000).
Lee, S.H. et al. v-ATPase V0 subunit d2–deficient mice exhibit impaired osteoclast fusion and increased bone formation. Nat. Med. 12, 1403–1409 (2006).
Hanash, S. Disease proteomics. Nature 422, 226–232 (2003).
Wyss, M. & Kaddurah-Daouk, R. Creatine and creatinine metabolism. Physiol. Rev. 80, 1107–1213 (2000).
Kuiper, J.W. et al. Creatine kinase–mediated ATP supply fuels actin-based events in phagocytosis. PLoS Biol. 6, e51 (2008).
Chellaiah, M.A. Regulation of actin ring formation by rho GTPases in osteoclasts. J. Biol. Chem. 280, 32930–32943 (2005).
Zhang, D. et al. The small GTP-binding protein, rho p21, is involved in bone resorption by regulating cytoskeletal organization in osteoclasts. J. Cell Sci. 108, 2285–2292 (1995).
Mahajan, V.B. et al. Creatine kinase, an ATP-generating enzyme, is required for thrombin receptor signaling to the cytoskeleton. Proc. Natl. Acad. Sci. USA 97, 12062–12067 (2000).
Sistermans, E.A. et al. Co-localization and functional coupling of creatine kinase B and gastric H+/K+-ATPase on the apical membrane and the tubulovesicular system of parietal cells. Biochem. J. 311, 445–451 (1995).
Jost, C.R. et al. Creatine kinase B–driven energy transfer in the brain is important for habituation and spatial learning behaviour, mossy fibre field size and determination of seizure susceptibility. Eur. J. Neurosci. 15, 1692–1706 (2002).
Dzeja, P.P., Terzic, A. & Wieringa, B. Phosphotransfer dynamics in skeletal muscle from creatine kinase gene-deleted mice. Mol. Cell. Biochem. 256–257, 13–27 (2004).
Crozatier, B. et al. Role of creatine kinase in cardiac excitation-contraction coupling: studies in creatine kinase–deficient mice. FASEB J. 16, 653–660 (2002).
Kaasik, A. et al. Energetic crosstalk between organelles: architectural integration of energy production and utilization. Circ. Res. 89, 153–159 (2001).
van Deursen, J. et al. Skeletal muscles of mice deficient in muscle creatine kinase lack burst activity. Cell 74, 621–631 (1993).
Lee, Y. et al. The ubiquitin-mediated degradation of Jak1 modulates osteoclastogenesis by limiting interferon-β–induced inhibitory signaling. Blood 111, 885–893 (2008).
Chang, E.J. et al. Elucidation of CPX-1 involvement in RANKL-induced osteoclastogenesis by a proteomics approach. FEBS Lett. 564, 166–170 (2004).
Ryu, J. et al. Sphingosine 1-phosphate as a regulator of osteoclast differentiation and osteoclast-osteoblast coupling. EMBO J. 25, 5840–5851 (2006).
Fernandez, R. & Malnic, G. H+ ATPase and Cl− interaction in regulation of MDCK cell pH. J. Membr. Biol. 163, 137–145 (1998).
Jimi, E. et al. Selective inhibition of NF-κB blocks osteoclastogenesis and prevents inflammatory bone destruction in vivo. Nat. Med. 10, 617–624 (2004).
Grey, A. et al. A role for interleukin-6 in parathyroid hormone-induced bone resorption in vivo. Endocrinology 140, 4683–4690 (1999).
Chang, E.J. et al. Hyaluronan inhibits osteoclast differentiation via Toll-like receptor 4. J. Cell Sci. 120, 166–176 (2007).
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
Authors and Affiliations
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
Supplementary information
Supplementary Text and Figures
Supplementary Figs. 1–8 and Supplementary Methods (PDF 4931 kb)
Rights and permissions
About this article
Cite this article
Chang, EJ., Ha, J., Oerlemans, F. et al. Brain-type creatine kinase has a crucial role in osteoclast-mediated bone resorption. Nat Med 14, 966–972 (2008). https://doi.org/10.1038/nm.1860
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nm.1860
This article is cited by
-
Concomitant induction of SLIT3 and microRNA-218–2 in macrophages by toll-like receptor 4 activation limits osteoclast commitment
Cell Communication and Signaling (2023)
-
The dynactin subunit DCTN1 controls osteoclastogenesis via the Cdc42/PAK2 pathway
Experimental & Molecular Medicine (2020)
-
S100A4 released from highly bone-metastatic breast cancer cells plays a critical role in osteolysis
Bone Research (2019)
-
Salt-inducible kinase 1 regulates bone anabolism via the CRTC1–CREB–Id1 axis
Cell Death & Disease (2019)
-
Interleukin-32 Gamma Stimulates Bone Formation by Increasing miR-29a in Osteoblastic Cells and Prevents the Development of Osteoporosis
Scientific Reports (2017)