The immunosuppressant rapamycin, alone or with transforming growth factor-β, enhances osteoclast differentiation of RAW264.7 Monocyte-macrophage cells in the presence of RANK-ligand

C. Shui, B. L. Riggs, Sundeep Khosla

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

Immunosuppressant therapy is known to cause bone loss. Since this may partly result from direct effects on osteoclast development, we investigated whether cyclosporin A (CsA), rapamycin, or FK506 affect osteoclastic differentiation of RAW264.7 monocytic cells induced by RANK-ligand (RANKL). Furthermore, since the rapamycin receptor protein binds transforming growth factor β (TGF-β) receptors, and TGF-β enhances osteoclastogenesis induced by RANKL, we also examined potential synergistic effects of rapamycin and TGF-β1. Rapamycin inhibited cell proliferation and stimulated tartrate-resistant acid phosphatase (TRAP) activity of RAW cells in a dose-dependent manner. At the optimal concentration of 10 ng/ml, it increased the number of TRAP+ multinucleated cells (MNC) more than 20-fold and enhanced the expression of TRAP and calcitonin receptor (CTR) mRNAs 2.1- and 10-fold, respectively. CsA, at 125-2000 ng/ml, similarly inhibited proliferation, but at high doses (1000-2000 ng/ml) it decreased TRAP activity, TRAP+MNC formation, and the expression of TRAP and CTR mRNAs. FK506 had no effect on cell proliferation or TRAP activity at concentrations up to 2000 ng/ml; however, like CsA, 1000 ng/ml FK506 inhibited TRAP+MNC formation and the expression of TRAP and CTR mRNAs. The combination of rapamycin (10 ng/ml) and TGF-β1(1 ng/ml) increased TRAP+MNC 3.1- and 6.9-fold as compared with rapamycin or TGF-β1 alone, respectively, and enhanced CTR mRNA expression induced by TGF-β1 by 1.9-fold. Rapamycin also increased osteoclastic resorption activity by 6.5-fold compared with control, and this was enhanced further by the addition of TGF-β by 3-fold, compared with rapamycin alone. These data thus indicate that rapamycin, alone or in synergy with TGF-β, directly enhances osteoclastogenesis and may affect bone metabolism in vivo after long-term use.

Original languageEnglish (US)
Pages (from-to)437-446
Number of pages10
JournalCalcified Tissue International
Volume71
Issue number5
DOIs
StatePublished - Nov 1 2002

Fingerprint

RANK Ligand
Transforming Growth Factors
Osteoclasts
Sirolimus
Immunosuppressive Agents
Monocytes
Macrophages
Calcitonin Receptors
Tacrolimus
Cyclosporine
Messenger RNA
Osteogenesis
Cell Proliferation
Tartrate-Resistant Acid Phosphatase
Bone and Bones
Growth Factor Receptors

Keywords

  • Immunosuppressant
  • Osteoclast
  • RANK-ligand
  • Transforming growth factor β

ASJC Scopus subject areas

  • Endocrinology

Cite this

@article{c21ca37cb1ef4edab404a76cc0567b19,
title = "The immunosuppressant rapamycin, alone or with transforming growth factor-β, enhances osteoclast differentiation of RAW264.7 Monocyte-macrophage cells in the presence of RANK-ligand",
abstract = "Immunosuppressant therapy is known to cause bone loss. Since this may partly result from direct effects on osteoclast development, we investigated whether cyclosporin A (CsA), rapamycin, or FK506 affect osteoclastic differentiation of RAW264.7 monocytic cells induced by RANK-ligand (RANKL). Furthermore, since the rapamycin receptor protein binds transforming growth factor β (TGF-β) receptors, and TGF-β enhances osteoclastogenesis induced by RANKL, we also examined potential synergistic effects of rapamycin and TGF-β1. Rapamycin inhibited cell proliferation and stimulated tartrate-resistant acid phosphatase (TRAP) activity of RAW cells in a dose-dependent manner. At the optimal concentration of 10 ng/ml, it increased the number of TRAP+ multinucleated cells (MNC) more than 20-fold and enhanced the expression of TRAP and calcitonin receptor (CTR) mRNAs 2.1- and 10-fold, respectively. CsA, at 125-2000 ng/ml, similarly inhibited proliferation, but at high doses (1000-2000 ng/ml) it decreased TRAP activity, TRAP+MNC formation, and the expression of TRAP and CTR mRNAs. FK506 had no effect on cell proliferation or TRAP activity at concentrations up to 2000 ng/ml; however, like CsA, 1000 ng/ml FK506 inhibited TRAP+MNC formation and the expression of TRAP and CTR mRNAs. The combination of rapamycin (10 ng/ml) and TGF-β1(1 ng/ml) increased TRAP+MNC 3.1- and 6.9-fold as compared with rapamycin or TGF-β1 alone, respectively, and enhanced CTR mRNA expression induced by TGF-β1 by 1.9-fold. Rapamycin also increased osteoclastic resorption activity by 6.5-fold compared with control, and this was enhanced further by the addition of TGF-β by 3-fold, compared with rapamycin alone. These data thus indicate that rapamycin, alone or in synergy with TGF-β, directly enhances osteoclastogenesis and may affect bone metabolism in vivo after long-term use.",
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author = "C. Shui and Riggs, {B. L.} and Sundeep Khosla",
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T1 - The immunosuppressant rapamycin, alone or with transforming growth factor-β, enhances osteoclast differentiation of RAW264.7 Monocyte-macrophage cells in the presence of RANK-ligand

AU - Shui, C.

AU - Riggs, B. L.

AU - Khosla, Sundeep

PY - 2002/11/1

Y1 - 2002/11/1

N2 - Immunosuppressant therapy is known to cause bone loss. Since this may partly result from direct effects on osteoclast development, we investigated whether cyclosporin A (CsA), rapamycin, or FK506 affect osteoclastic differentiation of RAW264.7 monocytic cells induced by RANK-ligand (RANKL). Furthermore, since the rapamycin receptor protein binds transforming growth factor β (TGF-β) receptors, and TGF-β enhances osteoclastogenesis induced by RANKL, we also examined potential synergistic effects of rapamycin and TGF-β1. Rapamycin inhibited cell proliferation and stimulated tartrate-resistant acid phosphatase (TRAP) activity of RAW cells in a dose-dependent manner. At the optimal concentration of 10 ng/ml, it increased the number of TRAP+ multinucleated cells (MNC) more than 20-fold and enhanced the expression of TRAP and calcitonin receptor (CTR) mRNAs 2.1- and 10-fold, respectively. CsA, at 125-2000 ng/ml, similarly inhibited proliferation, but at high doses (1000-2000 ng/ml) it decreased TRAP activity, TRAP+MNC formation, and the expression of TRAP and CTR mRNAs. FK506 had no effect on cell proliferation or TRAP activity at concentrations up to 2000 ng/ml; however, like CsA, 1000 ng/ml FK506 inhibited TRAP+MNC formation and the expression of TRAP and CTR mRNAs. The combination of rapamycin (10 ng/ml) and TGF-β1(1 ng/ml) increased TRAP+MNC 3.1- and 6.9-fold as compared with rapamycin or TGF-β1 alone, respectively, and enhanced CTR mRNA expression induced by TGF-β1 by 1.9-fold. Rapamycin also increased osteoclastic resorption activity by 6.5-fold compared with control, and this was enhanced further by the addition of TGF-β by 3-fold, compared with rapamycin alone. These data thus indicate that rapamycin, alone or in synergy with TGF-β, directly enhances osteoclastogenesis and may affect bone metabolism in vivo after long-term use.

AB - Immunosuppressant therapy is known to cause bone loss. Since this may partly result from direct effects on osteoclast development, we investigated whether cyclosporin A (CsA), rapamycin, or FK506 affect osteoclastic differentiation of RAW264.7 monocytic cells induced by RANK-ligand (RANKL). Furthermore, since the rapamycin receptor protein binds transforming growth factor β (TGF-β) receptors, and TGF-β enhances osteoclastogenesis induced by RANKL, we also examined potential synergistic effects of rapamycin and TGF-β1. Rapamycin inhibited cell proliferation and stimulated tartrate-resistant acid phosphatase (TRAP) activity of RAW cells in a dose-dependent manner. At the optimal concentration of 10 ng/ml, it increased the number of TRAP+ multinucleated cells (MNC) more than 20-fold and enhanced the expression of TRAP and calcitonin receptor (CTR) mRNAs 2.1- and 10-fold, respectively. CsA, at 125-2000 ng/ml, similarly inhibited proliferation, but at high doses (1000-2000 ng/ml) it decreased TRAP activity, TRAP+MNC formation, and the expression of TRAP and CTR mRNAs. FK506 had no effect on cell proliferation or TRAP activity at concentrations up to 2000 ng/ml; however, like CsA, 1000 ng/ml FK506 inhibited TRAP+MNC formation and the expression of TRAP and CTR mRNAs. The combination of rapamycin (10 ng/ml) and TGF-β1(1 ng/ml) increased TRAP+MNC 3.1- and 6.9-fold as compared with rapamycin or TGF-β1 alone, respectively, and enhanced CTR mRNA expression induced by TGF-β1 by 1.9-fold. Rapamycin also increased osteoclastic resorption activity by 6.5-fold compared with control, and this was enhanced further by the addition of TGF-β by 3-fold, compared with rapamycin alone. These data thus indicate that rapamycin, alone or in synergy with TGF-β, directly enhances osteoclastogenesis and may affect bone metabolism in vivo after long-term use.

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KW - Transforming growth factor β

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