Journal of Bone and Mineral Metabolism

, Volume 36, Issue 4, pp 399–409 | Cite as

Bone metabolism markers and angiogenic cytokines as regulators of human hematopoietic stem cell mobilization

  • Pantelis Tsirkinidis
  • Evangelos Terpos
  • Georgios Boutsikas
  • Athanasios Papatheodorou
  • Konstantinos Anargyrou
  • Eleni Lalou
  • Aglaia Dimitrakopoulou
  • Christina Kalpadakis
  • Konstantinos Konstantopoulos
  • Marina Siakantaris
  • Panayiotis Panayiotidis
  • Gerassimos Pangalis
  • Marie-Christine Kyrtsonis
  • Theodoros Vassilakopoulos
  • Maria K. AngelopoulouEmail author
Original Article


Hematopoietic stem cell (HSC) mobilization involves cleavage of ligands between HSC and niche components. However, there are scarce data regarding the role of bone cells in human HSC mobilization. We studied biochemical markers of bone metabolism and angiogenic cytokines during HSC mobilization in 46 patients’ sera with lymphoma and multiple myeloma, by ELISA. Significant changes between pre-mobilization and collection samples were found: (1) Bone alkaline phosphatase (BALP) increased, indicating augmentation of bone formation; (2) Receptor activator of Nf-κB ligand/osteoprotegerin ratio (RANKL/OPG) increased, showing osteoclastic differentiation and survival; however, there was no evidence of increased osteoclastic activity; and (3) Angiopoietin-1/Angiopoietin-2 ratio (ANGP-1/ANGP-2) decreased, consistent with vessel destabilization. Poor mobilizers had significantly higher carboxy-terminal telopeptide of collagen type I (CTX) and lower ANGP-1 at pre-mobilization samples, compared to good ones. CTX, amino-terminal telopeptide of collagen type I (NTX) and ANGP-1 pre-mobilization levels correlated significantly with circulating CD34+ peak cell counts. Our results indicate that bone formation and vessel destabilization are the two major events during human HSC mobilization. Osteoblasts seem to be the orchestrating cells, while osteoclasts are stimulated but not fully active. Moreover, ANGP-1, CTX and NTX may serve as predictors of poor mobilization.


Bone markers Angiopoietins Hematopoietic stem cells Mobilization Bone cells 


Compliance with ethical standards

Conflict of interest

All the authors declare that they have no conflict of interests related to the submitted paper.


  1. 1.
    Bonig H, Papayannopoulou T (2012) Mobilization of hematopoietic stem/progenitor cells: general principles and molecular mechanisms. Methods Mol Biol 904:1–14PubMedPubMedCentralGoogle Scholar
  2. 2.
    Angelopoulou MK, Tsirkinidis P, Boutsikas G, Vassilakopoulos TP, Tsirigotis P (2014) New insights in the mobilization of hematopoietic stem cells in lymphoma and multiple myeloma patients. Biomed Res Int 835138Google Scholar
  3. 3.
    Winkler IG, Sims NA, Pettit AR, Barbier V, Nowlan B, Helwani F, Poulton IJ, van Rooijen N, Alexander KA, Raggatt LJ, Lévesque JP (2010) Bone marrow macrophages maintain hematopoietic stem cell (HSC) niches and their depletion mobilizes HSCs. Blood 116:4815–4828CrossRefPubMedGoogle Scholar
  4. 4.
    Semerad CL, Christopher MJ, Liu F, Short B, Simmons PJ, Winkler I, Levesque JP, Chappel J, Ross FP, Link DC (2005) G-CSF potently inhibits osteoblast activity and CXCL12 mRNA expression in the bone marrow. Blood 106:3020–3027CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Christopher MJ, Liu F, Hilton MJ, Long F, Link DC (2009) Suppression of CXCL12 production by bone marrow osteoblasts is a common and critical pathway for cytokine-induced mobilization. Blood 114:1331–1339CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Ehninger A, Trumpp A (2011) The bone marrow stem cell niche grows up: mesenchymal stem cells and macrophages move in. J Exp Med 208:421–428CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Olivieri A, Marchetti M, Lemoli R, Tarella C, Iacone A, Lanza F, Rambaldi A, Bosi A; Italian Group for Stem Cell Transplantation (2012) Proposed definition of ‘poor’ mobilizers in lymphoma and multiple myeloma: an analytic hierarchy process by ad hoc working group Gruppo Italiano Trapianto di Midollo Osseo. BMT 47:342–351Google Scholar
  8. 8.
    Watts NB (1999) Clinical utility of biochemical markers of bone remodeling. Clin Chem 45:1359–1368PubMedGoogle Scholar
  9. 9.
    Calvo MS, Eyre DR, Gundberg CM (1996) Molecular basis and clinical application of biological markers of bone turnover. Endocr Rev 17:333–368PubMedGoogle Scholar
  10. 10.
    Halleen JM, Ylipahkala H, Alatalo SL, Janckila AJ, Heikkinen JE, Suominen H, Cheng S, Väänänen HK (2002) Serum tartrate-resistant acid phosphatase 5b, but not 5a, correlates with other markers of bone turnover and bone mineral density. Calcif Tissue Int 71:20–25CrossRefPubMedGoogle Scholar
  11. 11.
    Orimo H (2010) The mechanism of mineralization and the role of alkaline phosphatase in health and disease. J Nippon Med Sch 77:4–12CrossRefPubMedGoogle Scholar
  12. 12.
    Neve A, Corrado A, Cantatore FP (2013) Osteocalcin: skeletal and extra-skeletal effects. J Cell Physiol 228:1149–1153CrossRefPubMedGoogle Scholar
  13. 13.
    Sodek J, Gnss B, McKee MD (2000) Osteopontin. Crit Rev Oral Biol Med 11:279–303CrossRefPubMedGoogle Scholar
  14. 14.
    Pinzone JJ, Hall BM, Thudi NK, Vonau M, Qiang YW, Rosol TJ, Shaughnessy JD Jr (2009) The role of Dickkopf-1 in bone development, homeostasis, and disease. Blood 113:517–525CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Hofbauer LC (1999) Osteoprotegerin ligand and osteoprotegerin: novel implications for osteoclast biology and bone metabolism. Eur J Endocrinol 141:195–210CrossRefPubMedGoogle Scholar
  16. 16.
    Pfaff D, Fiedler U, Augustin HG (2006) Emerging roles of the Angiopoietin-Tie and the ephrin-Eph systems as regulators of cell trafficking. J Leukoc Biol 80:719–726CrossRefPubMedGoogle Scholar
  17. 17.
    Tello-Montoliu A, Patel JV, Lip GY (2006) Angiogenin: a review of the pathophysiology and potential clinical applications. J Thromb Haemost 4:1864–1874CrossRefPubMedGoogle Scholar
  18. 18.
    Shin HI, Divieti P, Sims NA, Kobayashi T, Miao D, Karaplis AC, Baron R, Bringhurst R, Kronenberg HM (2004) Gp130-mediated signaling is necessary for normal osteoblastic function in vivo and in vitro. Endocrinology 145:1376–1385CrossRefPubMedGoogle Scholar
  19. 19.
    Xu FF, Zhu H, Li XM, Yang F, Chen JD, Tang B, Sun HG, Chu YN, Zheng RX, Liu YL, Wang LS, Zhang Y (2014) Intercellular adhesion molecule-1 inhibits osteogenic differentiation of mesenchymal stem cells and impairs bio-scaffold-mediated bone regeneration in vivo. Tissue Eng Part A 20:2768–2782CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Kucia M, Reca R, Miekus K, Wanzeck J, Wojakowski W, Janowska-Wieczorek A, Ratajczak J, Ratajczak MZ (2005) Trafficking of normal stem cells and metastasis of cancer stem cells involve similar mechanisms: pivotal role of the SDF-1–CXCR4 axis. Stem Cells 23:879–894CrossRefPubMedGoogle Scholar
  21. 21.
    Sanderson RD, Yang Y (2008) Syndecan-1: a dynamic regulator of the myeloma microenvironment. Clin Exp Metastasis 25:149–159CrossRefPubMedGoogle Scholar
  22. 22.
    Kollet O, Dar A, Shivtiel S, Kalinkovich A, Lapid K, Sztainberg Y, Tesio M, Samstein RM, Goichberg P, Spiegel A, Elson A, Lapidot T (2006) Osteoclasts degrade endosteal components and promote mobilization of hematopoietic progenitor cells. Nat Med 12:657–664CrossRefPubMedGoogle Scholar
  23. 23.
    Drake FH, Dodds RA, James IE, Connor JR, Debouck C, Richardson S, Lee-Rykaczewski E, Coleman L, Rieman D, Barthlow R, Hastings G, Gowen M (1996) Cathepsin K, but not cathepsins B, L, or S, is abundantly expressed in human osteoclasts. J Biol Chem 271:12511–12516CrossRefPubMedGoogle Scholar
  24. 24.
    Takamatsu Y, Simmons PJ, Moore RJ, Morris HA, To LB, Levesque JP (1998) Osteoclast-mediated bone resorption is stimulated during short-term administration of granulocyte colony-stimulating factor but is not responsible for hematopoietic progenitor cell mobilization. Blood 92:3465–3473PubMedGoogle Scholar
  25. 25.
    Bergmann P, Body JJ, Boonen S, Boutsen Y, Devogelaer JP, Goemaere S, Kaufman JM, Reginster JY, Gangji V, Members of Advisory Board on Bone Markers (2009) Evidence-based guidelines for the use of biochemical markers of bone turnover in the selection and monitoring of bisphosphonate treatment in osteoporosis: a consensus document of the Belgian Bone Club. Int J Clin Pract 63:19–26CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Schwetz V, Pieber T, Obermayer-Pietsch B (2012) The endocrine role of the skeleton: background and clinical evidence. Eur J Endocrinol 166:959–967CrossRefPubMedGoogle Scholar
  27. 27.
    Li S, Zhai Q, Zou D, Meng H, Xie Z, Li C, Wang Y, Qi J, Cheng T, Qiu L (2013) A pivotal role of bone remodeling in granulocyte colony stimulating factor induced hematopoietic stem/progenitor cells mobilization. J Cell Physiol 228:1002–1009CrossRefPubMedGoogle Scholar
  28. 28.
    Tanaka M, Yujiri T, Tanaka Y, Mitani N, Tanimura A, Tanizawa Y (2012) Alteration of Dickkopf-1 and receptor activator of nuclear factor-kB ligand during PBSC mobilization in healthy donors by G-CSF [letter]. Bone Marrow Transplant 47:1143–1144CrossRefPubMedGoogle Scholar
  29. 29.
    Miyamoto K, Yoshida S, Kawasumi M, Hashimoto K, Kimura T et al (2011) Osteoclasts are dispensable for hematopoietic stem cell maintenance and mobilization. J Exp Med 208:2175–2181CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Szmigielska-Kaplon A, Krawczynska A, Czemerska M, Pluta A, Cebula-Obrzut B, Szmigielska K, Stępka K, Smolewski P, Robak T, Wierzbowska A (2015) Angiopoietins in hematopoietic stem cell mobilization in patients with haematological malignancies. Blood Transfus 13:102–108PubMedPubMedCentralGoogle Scholar
  31. 31.
    Anargyrou K, Terpos E, Vassilakopoulos TP, Pouli A, Sachanas S, Tzenou T, Masouridis S, Christoulas D, Angelopoulou MK, Dimitriadou EM, Kalpadakis C, Tsionos K, Panayiotidis P, Dimopoulos MA, Pangalis GA, Kyrtsonis MC; Greek Myeloma Study Group (2008) Normalization of the serum angiopoietin-1 to angiopoietin-2 ratio reflects response in refractory/resistant multiple myeloma patients treated with bortezomib. Haematologica 93:451–454Google Scholar
  32. 32.
    Terpos E, Anargyrou K, Katodritou E, Kastritis E, Papatheodorou A, Christoulas D, Pouli A, Michalis E, Delimpasi S, Gkotzamanidou M, Nikitas N, Koumoustiotis V, Margaritis D, Tsionos K, Stefanoudaki E, Meletis J, Zervas K, Dimopoulos MA; Greek Myeloma Study Group, Greece (2012) Circulating angiopoietin-1 to angiopoietin-2 ratio is an independent prognostic factor for survival in newly diagnosed patients with multiple myeloma who received therapy with novel antimyeloma agents. Int J Cancer 130:735–742Google Scholar
  33. 33.
    Tat SK, Pelletier JP, Velasco CR, Padrines M, Martel-Pelletier J (2009) New perspective in osteoarthritis: the OPG and RANKL system as a potential therapeutic target? Keio J Med 58:29–40CrossRefPubMedGoogle Scholar
  34. 34.
    Kelly T, Suva LJ, Nicks KM, MacLeod V, Sanderson RD (2010) Tumor-derived syndecan-1 mediates distal cross-talk with bone that enhances osteoclastogenesis. J Bone Miner Res 25:1295–1304CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Bonig H, Papayannopoulou T (2013) Hematopoietic stem cell mobilization: updated conceptual renditions. Leukemia 27:24–31CrossRefPubMedGoogle Scholar
  36. 36.
    Villalba S, Salvucci O, Aoki Y, De La Luz Sierra M, Gupta G, Davis D, Wyvill K, Little R, Yarchoan R, Tosato G (2003) Serum inactivation contributes to the failure of stromal-derived factor-1 to block HIV-I infection in vivo. J Leukoc Biol 74:880–888CrossRefPubMedGoogle Scholar
  37. 37.
    Coleman R, Brown J, Terpos E, Lipton A, Smith MR, Cook R, Major P (2008) Bone markers and their prognostic value in metastatic bone disease: clinical evidence and future directions. Cancer Treat Rev 34:629–639CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
    Ju HS, Leung S, Brown B, Stringer MA, Leigh S, Scherrer C, Shepard K, Jenkins D, Knudsen J, Cannon R (1997) Comparison of analytical performance and biological variability of three bone resorption assays. Clin Chem 43:1570–1576PubMedGoogle Scholar
  39. 39.
    van Bezooijen RL, Roelen BA, Visser A, van der Wee-Pals L, de Wilt E, Karperien M, Hamersma H, Papapoulos SE, ten Dijke P, Löwik CW (2004) Sclerostin is an osteocyte-expressed negative regulator of bone formation, but not a classical BMP antagonist. J Exp Med 199:805–814CrossRefPubMedPubMedCentralGoogle Scholar
  40. 40.
    Kobayashi Y, Uehara S, Udagawa N, Takahashi N (2016) Regulation of bone metabolism by Wnt signals. J Biochem 159:387–392CrossRefPubMedGoogle Scholar
  41. 41.
    Kamiya N, Kobayashi T, Mochida Y, Yu PB, Yamauchi M, Kronenberg HM, Mishina Y (2010) Wnt inhibitors Dkk1 and Sost are downstream targets of BMP signaling through the type IA receptor (BMPRIA) in osteoblasts. J Bone Miner Res 25:200–210CrossRefPubMedGoogle Scholar

Copyright information

© The Japanese Society for Bone and Mineral Research and Springer Japan KK 2017

Authors and Affiliations

  • Pantelis Tsirkinidis
    • 1
  • Evangelos Terpos
    • 2
  • Georgios Boutsikas
    • 3
  • Athanasios Papatheodorou
    • 4
  • Konstantinos Anargyrou
    • 3
  • Eleni Lalou
    • 5
  • Aglaia Dimitrakopoulou
    • 6
  • Christina Kalpadakis
    • 7
  • Konstantinos Konstantopoulos
    • 5
  • Marina Siakantaris
    • 8
  • Panayiotis Panayiotidis
    • 9
  • Gerassimos Pangalis
    • 10
  • Marie-Christine Kyrtsonis
    • 9
  • Theodoros Vassilakopoulos
    • 5
  • Maria K. Angelopoulou
    • 5
    Email author
  1. 1.Department of Hematology401 General Army Hospital of AthensAthensGreece
  2. 2.Department of Clinical Therapeutics, School of MedicineNational and Kapodistrian University of AthensAthensGreece
  3. 3.Department of Hematology251 General Air Force HospitalAthensGreece
  4. 4.Department of Medical Research251 General Air Force HospitalAthensGreece
  5. 5.Department of Hematology and Bone Marrow Transplantation, School of MedicineNational and Kapodistrian University of AthensAthensGreece
  6. 6.Department of Immunology Research and Flow Cytometry‘Laiko’ General Hospital of AthensAthensGreece
  7. 7.Department of Hematology, School of MedicineUniversity of CreteHerakleionGreece
  8. 8.1st Department of Internal Medicine, School of MedicineNational and Kapodistrian University of AthensAthensGreece
  9. 9.1st Propedeutic Department of Internal Medicine, School of MedicineNational and Kapodistrian University of AthensAthensGreece
  10. 10.Department of Hematology, Psychicon BranchAthens Medical CenterAthensGreece

Personalised recommendations