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Pathology & Oncology Research

, Volume 25, Issue 1, pp 409–419 | Cite as

In Vitro Study of the Effects of Denosumab on Giant Cell Tumor of Bone: Comparison with Zoledronic Acid

  • Isao ShibuyaEmail author
  • Masamichi TakamiEmail author
  • Arei Miyamoto
  • Akiko Karakawa
  • Akira Dezawa
  • Shigeru Nakamura
  • Ryutaro Kamijo
Original Article

Abstract

Giant cell tumor of bone (GCTB) is a locally aggressive primary bone tumor that contains numerous osteoclasts formed from marrow-derived precursors through receptor activator of nuclear factor κ-B ligand (RANKL), an osteoclast differentiation factor expressed in neoplastic cells of GCTB. Denosumab, a fully human monoclonal antibody targeting RANKL, has recently been used for the treatment of GCTB, and superior treatment effects have been reported. The aim of this work was to elucidate the mechanism of action of denosumab, and the differences between denosumab and zoledronic acid at the level of GCTB cells. We isolated GCTB cells from 3 patients and separated them into osteoclasts, osteoclast precursors and proliferating spindle-shaped stromal cells (the true neoplastic component), and examined the action of denosumab on differentiation, survival and bone resorption activity of osteoclasts. Denosumab and zoledronic acid inhibited osteoclast differentiation from mononuclear cells containing osteoclast precursors. Zoledronic acid inhibited osteoclast survival, whereas an inhibitory effect of denosumab on osteoclast survival was not observed. The inhibitory effect on bone resorption by both agents was confirmed in culture on dentin slices. Furthermore, zoledronic acid showed dose-dependent inhibition of cell growth of neoplastic cells whereas denosumab had no inhibitory effect on these cells. Denosumab has an inhibitory effect on osteoclast differentiation, but no inhibitory effects on survival of osteoclasts or growth of neoplastic cells in GCTBs.

Keywords

Giant cell tumor of bone Osteoclast Bisphosphonate Denosumab Bone tumor 

Notes

Acknowledgements

This work was supported by the Japan Society for the Promotion of Science (Grant Numbers 26293398 and 16 K20655) from the Ministry of Education, Culture, Sports, Science and Technology, Japan.

Author Contributions

Conception and design: Isao Shibuya, Masamichi Takami.

Analysis and interpretation of data: Isao Shibuya, Masamichi Takami, Arei Miyamoto, Akiko Karakawa.

Drafting the article or revising: Isao Shibuya, Masamichi Takami, Akira Dezawa, Shigeru Nakamura, Ryutaro Kamijo.

GUARANTOR: Isao Shibuya.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Ethical Approval

This study has been carried out with the approval of the ethics committee of Teikyo University School of Medicine (approval No: 12–001) and Showa University School of Dentistry (approval No: 2006–12). All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed Consent

Informed consent was obtained from all individual participants included in the study.

References

  1. 1.
    Wulling M, Engels C, Jesse N et al (2001) The nature of giant cell tumor of bone. J Cancer Res Clin Oncol 127:467–474CrossRefGoogle Scholar
  2. 2.
    Stewart MJ, Richardson TR (1952) Giant cell tumor of bone. J Bone Joint Surg (Am Vol) 34-A:272–286Google Scholar
  3. 3.
    Raskin KA, Schwab JH, Mankin HJ et al (2013) Giant cell tumor of bone. J Am Acad Orthop Surg 21:118–126CrossRefGoogle Scholar
  4. 4.
    Chambers TJ, Fuller K, McSheehy PM et al (1985) The effects of calcium regulating hormones on bone resorption by isolated human osteoclastoma cells. J Pathol 145:297–305CrossRefGoogle Scholar
  5. 5.
    Kanehisa J, Izumo T, Takeuchi M et al (1991) In vitro bone resorption by isolated multinucleated giant cells from giant cell tumour of bone: light and electron microscopic study. Virchows Arch A Pathol Anat Histopathol 419:327–338CrossRefGoogle Scholar
  6. 6.
    Drake FH, Dodds RA, James IE et al (1996) Cathepsin K, but not cathepsins B, L, or S, is abundantly expressed in human osteoclasts. J Biol Chem 271:12511–12516CrossRefGoogle Scholar
  7. 7.
    Huang L, Xu J, Wood DJ et al (2000) Gene expression of osteoprotegerin ligand, osteoprotegerin, and receptor activator of NF-kappaB in giant cell tumor of bone: possible involvement in tumor cell-induced osteoclast-like cell formation. Am J Pathol 156:761–767CrossRefGoogle Scholar
  8. 8.
    Joyner CJ, Quinn JM, Triffitt JT et al (1992) Phenotypic characterisation of mononuclear and multinucleated cells of giant cell tumour of bone. Bone Miner 16:37–48CrossRefGoogle Scholar
  9. 9.
    Lau YS, Sabokbar A, Gibbons CL et al (2005) Phenotypic and molecular studies of giant-cell tumors of bone and soft tissue. Hum Pathol 36:945–954CrossRefGoogle Scholar
  10. 10.
    Campanacci M, Baldini N, Boriani S et al (1987) Giant-cell tumor of bone. J Bone Joint Surg Am 69:106–114CrossRefGoogle Scholar
  11. 11.
    Arbeitsgemeinschaft K, Becker WT, Dohle J et al (2008) Local recurrence of giant cell tumor of bone after intralesional treatment with and without adjuvant therapy. J Bone Joint Surg Am 90:1060–1067CrossRefGoogle Scholar
  12. 12.
    Mori Y, Tsuchiya H, Karita M et al (2000) Malignant transformation of a giant cell tumor 25 years after initial treatment. Clin Orthop Relat Res 381:185–191CrossRefGoogle Scholar
  13. 13.
    Rock MG, Sim FH, Unni KK et al (1986) Secondary malignant giant-cell tumor of bone. Clinicopathological assessment of nineteen patients. J Bone Joint Surg Am 68:1073–1079CrossRefGoogle Scholar
  14. 14.
    Nascimento AG, Huvos AG, Marcove RC (1979) Primary malignant giant cell tumor of bone: a study of eight cases and review of the literature. Cancer 44:1393–1402CrossRefGoogle Scholar
  15. 15.
    Bertoni F, Present D, Sudanese A et al (1988) Giant-cell tumor of bone with pulmonary metastases. Six case reports and a review of the literature. Clin Orthop Relat Res 237:275–285Google Scholar
  16. 16.
    Kay RM, Eckardt JJ, Seeger LL et al (1994) Pulmonary metastasis of benign giant cell tumor of bone. Six histologically confirmed cases, including one of spontaneous regression. Clin Orthop Relat Res 302:219–230Google Scholar
  17. 17.
    Nakashima T, Hayashi M, Takayanagi H (2012) New insights into osteoclastogenic signaling mechanisms. Trends Endocrinol Metab 23:582–590CrossRefGoogle Scholar
  18. 18.
    Suda T, Takahashi N, Udagawa N et al (1999) Modulation of osteoclast differentiation and function by the new members of the tumor necrosis factor receptor and ligand families. Endocr Rev 20:345–357CrossRefGoogle Scholar
  19. 19.
    Blair HC, Athanasou NA (2004) Recent advances in osteoclast biology and pathological bone resorption. Histol Histopathol 19:189–199Google Scholar
  20. 20.
    Shiotani A, Takami M, Itoh K et al (2002) Regulation of osteoclast differentiation and function by receptor activator of NFkB ligand and osteoprotegerin. Anat Rec 268:137–146CrossRefGoogle Scholar
  21. 21.
    Blackley HR, Wunder JS, Davis AM et al (1999) Treatment of giant-cell tumors of long bones with curettage and bone-grafting. J Bone Joint Surg Am 81:811–820CrossRefGoogle Scholar
  22. 22.
    XC Y, Xu M, Song RX et al (2010) Long-term outcome of giant cell tumors of bone around the knee treated by en bloc resection of tumor and reconstruction with prosthesis. Orthop Surg 2:211–217CrossRefGoogle Scholar
  23. 23.
    Puthoor DK, Puthezhath K (2012) Management of giant cell tumor of bone: computerized tomography based selection strategy and approaching the lesion through the site of cortical break. Orthop Surg 4:76–82CrossRefGoogle Scholar
  24. 24.
    Singh AS, Chawla NS, Chawla SP (2015) Giant-cell tumor of bone: treatment options and role of denosumab. Biologics 9:69–74Google Scholar
  25. 25.
    Tse LF, Wong KC, Kumta SM et al (2008) Bisphosphonates reduce local recurrence in extremity giant cell tumor of bone: a case-control study. Bone 42:68–73CrossRefGoogle Scholar
  26. 26.
    Balke M, Campanacci L, Gebert C et al (2010) Bisphosphonate treatment of aggressive primary, recurrent and metastatic Giant cell tumour of bone. BMC Cancer 10:462CrossRefGoogle Scholar
  27. 27.
    Yang T, Zheng XF, Lin X et al (2013) Postoperative irrigation with bisphosphonates may reduce the recurrence of giant cell tumor of bone. Med Hypotheses 81:851–852CrossRefGoogle Scholar
  28. 28.
    Gouin F, Rochwerger AR, Di Marco A et al (2014) Adjuvant treatment with zoledronic acid after extensive curettage for giant cell tumours of bone. Eur J Cancer 50:2425–2431CrossRefGoogle Scholar
  29. 29.
    Bekker PJ, Holloway DL, Rasmussen AS et al (2004) A single-dose placebo-controlled study of AMG 162, a fully human monoclonal antibody to RANKL, in postmenopausal women. J Bone Miner Res 19:1059–1066CrossRefGoogle Scholar
  30. 30.
    Lacey DL, Boyle WJ, Simonet WS et al (2012) Bench to bedside: elucidation of the OPG-RANK-RANKL pathway and the development of denosumab. Nat Rev Drug Discov 11:401–419CrossRefGoogle Scholar
  31. 31.
    Thomas D, Henshaw R, Skubitz K et al (2010) Denosumab in patients with giant-cell tumour of bone: an open-label, phase 2 study. Lancet Oncol 11:275–280CrossRefGoogle Scholar
  32. 32.
    Lewiecki EM (2010) Clinical use of denosumab for the treatment for postmenopausal osteoporosis. Curr Med Res Opin 26:2807–2812CrossRefGoogle Scholar
  33. 33.
    Morgan T, Atkins GJ, Trivett MK et al (2005) Molecular profiling of giant cell tumor of bone and the osteoclastic localization of ligand for receptor activator of nuclear factor kappaB. Am J Pathol 167:117–128CrossRefGoogle Scholar
  34. 34.
    Roux S, Amazit L, Meduri G et al (2002) RANK (receptor activator of nuclear factor kappa B) and RANK ligand are expressed in giant cell tumors of bone. Am J Clin Pathol 117:210–216CrossRefGoogle Scholar
  35. 35.
    Thomas DM (2012) RANKL, denosumab, and giant cell tumor of bone. Curr Opin Oncol 24:397–403CrossRefGoogle Scholar
  36. 36.
    Hakozaki M, Tajino T, Yamada H et al (2014) Radiological and pathological characteristics of giant cell tumor of bone treated with denosumab. Diagn Pathol 9:111CrossRefGoogle Scholar
  37. 37.
    Yamagishi T, Kawashima H, Ogose A et al (2016) Disappearance of giant cells and presence of newly formed bone in the pulmonary metastasis of a sacral giant-cell tumor following denosumab treatment: a case report. Oncol Lett 11:243–246CrossRefGoogle Scholar
  38. 38.
    van Beek E, Pieterman E, Cohen L et al (1999) Farnesyl pyrophosphate synthase is the molecular target of nitrogen-containing bisphosphonates. Biochem Biophys Res Commun 264:108–111CrossRefGoogle Scholar
  39. 39.
    Martin M, Bell R, Bourgeois H et al (2012) Bone-related complications and quality of life in advanced breast cancer: results from a randomized phase III trial of denosumab versus zoledronic acid. Clin Cancer Res 18:4841–4849CrossRefGoogle Scholar
  40. 40.
    Lipton A, Fizazi K, Stopeck AT et al (2012) Superiority of denosumab to zoledronic acid for prevention of skeletal-related events: a combined analysis of 3 pivotal, randomised, phase 3 trials. Eur J Cancer 48:3082–3092CrossRefGoogle Scholar
  41. 41.
    Scagliotti GV, Hirsh V, Siena S et al (2012) Overall survival improvement in patients with lung cancer and bone metastases treated with denosumab versus zoledronic acid: subgroup analysis from a randomized phase 3 study. J Thorac Oncol 7:1823–1829CrossRefGoogle Scholar
  42. 42.
    Branstetter DG, Nelson SD, Manivel JC et al (2012) Denosumab induces tumor reduction and bone formation in patients with giant-cell tumor of bone. Clin Cancer Res 18:4415–4424CrossRefGoogle Scholar
  43. 43.
    Cheng YY, Huang L, Lee KM et al (2004) Bisphosphonates induce apoptosis of stromal tumor cells in giant cell tumor of bone. Calcif Tissue Int 75:71–77CrossRefGoogle Scholar
  44. 44.
    Nishisho T, Hanaoka N, Miyagi R et al (2015) Local administration of zoledronic acid for giant cell tumor of bone. Orthopedics 38:e25–e30CrossRefGoogle Scholar
  45. 45.
    Salerno M, Avnet S, Alberghini M et al (2008) Histogenetic characterization of giant cell tumor of bone. Clin Orthop Relat Res 466:2081–2091CrossRefGoogle Scholar
  46. 46.
    Avnet S, Salerno M, Zini N et al (2013) Sustained autocrine induction and impaired negative feedback of osteoclastogenesis in CD14(+) cells of giant cell tumor of bone. Am J Pathol 182:1357–1366CrossRefGoogle Scholar
  47. 47.
    Takami M, Takahashi N, Udagawa N et al (2000) Intracellular calcium and protein kinase C mediate expression of receptor activator of nuclear factor-kappaB ligand and osteoprotegerin in osteoblasts. Endocrinology 141:4711–4719CrossRefGoogle Scholar
  48. 48.
    Agarwal A, Larsen BT, Buadu LD et al (2013) Denosumab chemotherapy for recurrent giant-cell tumor of bone: a case report of neoadjuvant use enabling complete surgical resection. Case Rep Oncol Med 2013:496351Google Scholar
  49. 49.
    Akaike K, Suehara Y, Takagi T et al (2014) An eggshell-like mineralized recurrent lesion in the popliteal region after treatment of giant cell tumor of the bone with denosumab. Skelet Radiol 43:1767–1772CrossRefGoogle Scholar
  50. 50.
    Mattei TA, Ramos E, Rehman AA et al (2014) Sustained long-term complete regression of a giant cell tumor of the spine after treatment with denosumab. Spine J 14:e15–e21CrossRefGoogle Scholar
  51. 51.
    Atkins J, Kostakis P, Vincent C et al (2006) RANK expression as a cell surface marker of human osteoclast precursors in peripheral blood, bone marrow, and giant cell tumors of bone. J Bone Miner Res 21:1339–1349CrossRefGoogle Scholar
  52. 52.
    Liverani C, Mercatali L, Spadazzi C et al (2014) CSF-1 blockade impairs breast cancer osteoclastogenic potential in co-culture systems. Bone 66:214–222CrossRefGoogle Scholar
  53. 53.
    Cheng YY, Huang L, Kumta SM et al (2003) Cytochemical and ultrastructural changes in the osteoclast-like giant cells of giant cell tumor of bone following bisphosphonate administration. Ultrastruct Pathol 27:385–391CrossRefGoogle Scholar
  54. 54.
    Lau CP, Huang L, Wong KC et al (2013) Comparison of the anti-tumor effects of denosumab and zoledronic acid on the neopastic cells of giant cell tumor of bone. Connect Tissue Res 54:439–449CrossRefGoogle Scholar
  55. 55.
    Ueda T, Morioka H, Nishida Y et al (2015) Objective tumor response to denosumab in patients with giant cell tumor of bone: a multicenter phase II trial. Ann Oncol 26:2149–2154CrossRefGoogle Scholar
  56. 56.
    Akiyama T, Choong PF, Dass CR (2010) RANK-fc inhibits malignancy via inhibiting ERK activation and evoking caspase-3-mediated anoikis in human osteosarcoma cells. Clin Exp Metastasis 27:207–215CrossRefGoogle Scholar
  57. 57.
    James IE, Dodds RA, Olivera DL et al (1996) Human osteoclastoma-derived stromal cells: correlation of the ability to form mineralized nodules in vitro with formation of bone in vivo. J Bone Miner Res 11:1453–1460CrossRefGoogle Scholar
  58. 58.
    Murata A, Fujita T, Kawahara N et al (2005) Osteoblast lineage properties in giant cell tumors of bone. J Orthop Sci 10:581–588CrossRefGoogle Scholar

Copyright information

© Arányi Lajos Foundation 2017

Authors and Affiliations

  • Isao Shibuya
    • 1
    • 2
    Email author
  • Masamichi Takami
    • 3
    Email author
  • Arei Miyamoto
    • 1
  • Akiko Karakawa
    • 3
  • Akira Dezawa
    • 2
  • Shigeru Nakamura
    • 2
  • Ryutaro Kamijo
    • 1
  1. 1.Department of BiochemistryShowa University School of DentistryTokyoJapan
  2. 2.Department of Orthopaedic SurgeryTeikyo University Mizonokuchi HospitalKawasakiJapan
  3. 3.Department of PharmacologyShowa University School of DentistryTokyoJapan

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