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Molecular Medicine

, Volume 23, Issue 1, pp 57–69 | Cite as

α-1 Antitrypsin Inhibits RANKL-induced Osteoclast Formation and Functions

  • Mohammad Ahsanul Akbar
  • David Nardo
  • Mong-Jen Chen
  • Ahmed S. Elshikha
  • Rubina Ahamed
  • Eslam M. Elsayed
  • Claire Bigot
  • L. Shannon Holliday
  • Sihong Song
Research Article

Abstract

Osteoporosis is a global public health problem affecting more than 200 million people worldwide. We previously showed that treatment with α-1 antitrypsin (AAT), a multifunctional protein with antiinflammatory properties, mitigated bone loss in an ovariectomized mouse model. However, the underlying mechanisms of the protective effect of AAT on bone tissue are largely unknown. In this study, we investigated the effect of AAT on osteoclast formation and function in vitro. Our results showed that AAT dose-dependently inhibited the formation of receptor activator of nuclear factor κB ligand (RANKL)-induced osteoclasts derived from mouse bone marrow macrophage/monocyte (BMM) lineage cells and the RAW 264.7 murine macrophage cell line. To elucidate the possible mechanisms underlying this inhibition, we tested the effect of AAT on the gene expression of cell surface molecules, transcription factors and cytokines associated with osteoclast formation. We showed that AAT inhibited macrophage colony-stimulating factor (M-CSF)-induced cell surface RANK expression in osteoclast precursor cells. In addition, AAT inhibited RANKL-induced TNF-α production, cell surface CD9 expression and dendritic cell-specific transmembrane protein (DC-STAMP) gene expression. Importantly, AAT treatment significantly inhibited osteoclast-associated mineral resorption. Together, these results uncover new mechanisms for the protective effects of AAT and strongly support the notion that AAT has therapeutic potential for the treatment of osteoporosis.

Notes

Acknowledgments

EME is a visiting scholar from Zagazig University and is supported by a scholarship from the Egyptian government. We thank Dr. Jay Cao (US Department of Agriculture, Agriculture Research Service’s Grand Forks Human Nutrition Research Center) for his assistance with part of the gene expression studies and suggestions regarding induction of osteoclast formation.

This work was supported by a grant from the University of Florida.

Supplementary material

10020_2017_2301057_MOESM1_ESM.pdf (1.9 mb)
Supplementary material, approximately 1.88 MB.

References

  1. 1.
    Baron R, Hesse E. (2012) Update on bone anabolics in osteoporosis treatment: rationale, current status, and perspectives. J. Clin. Endocrinol. Metab. 97:311–25.CrossRefGoogle Scholar
  2. 2.
    Lee JW, et al. (2010) Alisol-B, a novel phytosteroid, suppresses the RANKL-induced osteoclast formation and prevents bone loss in mice. Biochem. Pharmacol. 80:352–61.CrossRefGoogle Scholar
  3. 3.
    Troen BR. (2003) Molecular mechanisms underlying osteoclast formation and activation. Exp. Gerontol. 38:605–14.CrossRefGoogle Scholar
  4. 4.
    Nanes MS, Kallen CB (2014) Osteoporosis. Sem. Nucl. Med. 44:439–50.CrossRefGoogle Scholar
  5. 5.
    Kimachi K, Kajiya H, Nakayama S, Ikebe T, Okabe K. (2011) Zoledronic acid inhibits RANK expression and migration of osteoclast precursors during osteoclastogenesis. Naunyn Schmiedebergs Arch. Pharmacol. 383:297–308.CrossRefGoogle Scholar
  6. 6.
    Suda T, Takahashi N, Martin TJ. (1992) Modulation of osteoclast differentiation. Endocr. Rev. 13:66–80.PubMedGoogle Scholar
  7. 7.
    Lacey D, et al. (1998) Osteoprotegerin ligand is a cytokine that regulates osteoclast differentiation and activation. Cell. 93:165–76.CrossRefGoogle Scholar
  8. 8.
    Yasuda H, et al. (1998) Osteoclast differentiation factor is a ligand for osteoprotegerin osteoclastogenesis inhibitory factor and is identical to TRANCE/RANKL. Proc. Natl. Acad. Sci. USA. 95:3597–3602.CrossRefGoogle Scholar
  9. 9.
    Arai F, et al. (1999) Commitment and differentiation of osteoclast precursor cells by the sequential expression of c-Fms and receptor activator of nuclear factor kappaB (RANK) receptors. J. Exp. Med. 190:1741–54.CrossRefGoogle Scholar
  10. 10.
    Ishida N, et al. (2002) Large Scale Gene Expression Analysis of Osteoclastogenesis in Vitro and Elucidation of NFAT2 as a Key Regulator. J. Biol. Chem. 277:41147–56.CrossRefGoogle Scholar
  11. 11.
    Johnson RS, Spiegelman BM, Papaioannou V. (1992) Pleiotropic effects of a null mutation in the c-fos proto-oncogene. Cell. 71:577–86.CrossRefGoogle Scholar
  12. 12.
    Takayanagi H, et al. (2002) Induction and activation of the transcription factor NFATc1 (NFAT2) integrate RANKL signaling in terminal 1differentiation of osteoclasts. Dev. Cell. 3:889–901.CrossRefGoogle Scholar
  13. 13.
    Xing L, Xiu Y, Boyce BF. (2012) Osteoclast fusion and regulation by RANKL-dependent and independent factors. World J. Orthop. 3:212–22.CrossRefGoogle Scholar
  14. 14.
    Kukita T, et al. (2004) RANKL-induced DC-STAMP is essential for osteoclastogenesis. J. Exp. Med. 200:941–46.CrossRefGoogle Scholar
  15. 15.
    Burgess TL, et al. (1999) The ligand for osteoprotegerin (OPGL) directly activates mature osteoclasts. J. Cell Biol. 145:527–38.CrossRefGoogle Scholar
  16. 16.
    Blair HC, Teitelbaum SL, Ghiselli R, Gluck S. (1989) Osteoclastic bone resorption by a polarized vacuolar proton pump. Science. 245:855–57.CrossRefGoogle Scholar
  17. 17.
    Boyle WJ, Simonet WS, Lacey DL. (2003) Osteoclast differentiation and activation. Nature. 423:337–42.CrossRefGoogle Scholar
  18. 18.
    Wada T, Nakashima T, Hiroshi N, Penninger JM. (2006) RANKL-RANK signaling in osteoclastogenesis and bone disease. Trends Mol. Med. 12:17–25.CrossRefGoogle Scholar
  19. 19.
    Mundy GR. (2007) Osteoporosis and inflammation. Nutr. Rev. 65:S147–51.CrossRefGoogle Scholar
  20. 20.
    Romas E, Gillespie MT. (2006) Inflammation-induced bone loss: can it be prevented? Rheum. Dis. Clin. North Am. 32:759–73.CrossRefGoogle Scholar
  21. 21.
    Janciauskiene SM, et al. (2011) The discovery of alpha1-antitrypsin and its role in health and disease. Respir. Med. 105:1129–39.CrossRefGoogle Scholar
  22. 22.
    Lu YQ et al. (2006) Alpha(1)-antitrypsin gene therapy modulates cellular immunity and efficiently prevents type 1 diabetes in nonobese diabetic mice. Hum. Gene Ther. 17:625–34.CrossRefGoogle Scholar
  23. 23.
    Ma H, et al. (2010) Intradermal alpha1-antitrypsin therapy avoids fatal anaphylaxis, prevents type 1 diabetes and reverses hyperglycaemia in the NOD mouse model of the disease. Diabetologia. 53:2198–2204.CrossRefGoogle Scholar
  24. 24.
    Song S, et al. (2004) Recombinant adeno-associated virus-mediated alpha-1 antitrypsin gene therapy prevents type I diabetes in NOD mice. Gene Ther. 11:181–86.CrossRefGoogle Scholar
  25. 25.
    Grimstein C, et al. (2011) Alpha-1 antitrypsin protein and gene therapies decrease autoimmunity and delay arthritis development in mouse model. J. Transl. Med. 9:21.CrossRefGoogle Scholar
  26. 26.
    Grimstein C, et al. (2010) Combination of alpha-1 antitrypsin and doxycycline suppresses collagen-induced arthritis. J. Gene Med. 12:35–44.CrossRefGoogle Scholar
  27. 27.
    Shapiro L, Pott GB, Ralston AH. (2001) Alpha-1-antitrypsin inhibits human immunodeficiency virus type 1. FASEB J. 15:115–22.CrossRefGoogle Scholar
  28. 28.
    Akbar MA, et al (2016) Alpha-1 Antitrypsin Gene Therapy Ameliorates Bone Loss in Ovariectomy-Induced Osteoporosis Mouse Model. Hum. Gene Ther. 27:679–86.CrossRefGoogle Scholar
  29. 29.
    Akbar MA, et al. (2016) Transplantation of Adipose Tissue-Derived Mesenchymal Stem Cell (ATMSC) Expressing Alpha-1 Antitrypsin Reduces Bone Loss in Ovariectomized Osteoporosis Mice. Hum. Gene Ther. 28:179–89.CrossRefGoogle Scholar
  30. 30.
    Takahashi N, Udagawa N, Kobayashi Y, Suda T. (2007) Generation of osteoclasts in vitro, and assay of osteoclast activity. Methods Mol. Med. 135:285–301.CrossRefGoogle Scholar
  31. 31.
    Collin-Osdoby P, Yu X, Zheng H, Osdoby P. (2003) RANKL-mediated osteoclast formation from murine RAW 264.7 cells. Methods Mol. Med. 80:153–66.PubMedGoogle Scholar
  32. 32.
    Bergin DA, et al. (2014) The circulating proteinase inhibitor alpha-1 antitrypsin regulates neutrophil degranulation and autoimmunity. Sci. Transl. Med. 6:217ra211.CrossRefGoogle Scholar
  33. 33.
    Zou W, Hakim I, Tschoep K, Endres S, Bar-Shavit Z. (2001) Tumor necrosis factor-alpha mediates RANK ligand stimulation of osteoclast differentiation by an autocrine mechanism. J. Cell. Biochem. 83:70–83.CrossRefGoogle Scholar
  34. 34.
    Arai A, et al. (2012) Fos plays an essential role in the upregulation of RANK expression in osteoclast precursors within the bone microenvironment. J. Cell Sci. 125:2910–17.CrossRefGoogle Scholar
  35. 35.
    Petrache I, Hajjar J, Campos M. (2009) Safety and efficacy of alpha-1-antitrypsin augmentation therapy in the treatment of patients with alpha-1-antitrypsin deficiency. Biologics. 3:193–204.PubMedPubMedCentralGoogle Scholar
  36. 36.
    Mohanka M, Khemasuwan D, Stoller JK. (2012) A review of augmentation therapy for alpha-1 antitrypsin deficiency. Expert Opin. Biol. Ther. 12:685–700.CrossRefGoogle Scholar
  37. 37.
    Hough F, et al. (2014) The safety of osteoporosis medication. S. Afr. Med. J. 104:279–82.CrossRefGoogle Scholar
  38. 38.
    Zou W, Amcheslavsky A, Takeshita S, Drissi H, Bar-Shavit Z. (2005) TNF-α expression is transcriptionally regulated by RANK ligand. J. Cell. Physiol. 202:371–78.CrossRefGoogle Scholar
  39. 39.
    Bergin DA, et al. (2010) alpha-1 Antitrypsin regulates human neutrophil chemotaxis induced by soluble immune complexes and IL-8. J. Clin. Invest. 120:4236–50.CrossRefGoogle Scholar
  40. 40.
    Zhang B, et al. (2007) α1-Antitrypsin protects β-cells from apoptosis. Diabetes. 56:1316–23.CrossRefGoogle Scholar
  41. 41.
    Yao GQ, Itokawa T, Paliwal I, Insogna K. (2005) CSF-1 induces fos gene transcription and activates the transcription factor Elk-1 in mature osteoclasts. Calcif. Tissue Int. 76:371–78.CrossRefGoogle Scholar
  42. 42.
    Song I, et al. (2009) Regulatory mechanism of NFATc1 in RANKL-induced osteoclast activation. FEBS Lett. 583:2435–40.CrossRefGoogle Scholar

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Authors and Affiliations

  • Mohammad Ahsanul Akbar
    • 1
  • David Nardo
    • 1
  • Mong-Jen Chen
    • 1
  • Ahmed S. Elshikha
    • 1
  • Rubina Ahamed
    • 1
  • Eslam M. Elsayed
    • 1
    • 4
  • Claire Bigot
    • 1
  • L. Shannon Holliday
    • 2
    • 3
  • Sihong Song
    • 1
    • 5
  1. 1.Department of Pharmaceutics, College of PharmacyUniversity of FloridaGainesvilleUSA
  2. 2.Department of Orthodontics, College of DentistryUniversity of FloridaGainesvilleUSA
  3. 3.Department of Anatomy and Cell Biology, College of MedicineUniversity of FloridaGainesvilleUSA
  4. 4.Department of Microbiology and Immunology, Faculty of PharmacyZagazig UniversityCairoEgypt
  5. 5.GainesvilleUSA

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