Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Myeloma

HOXB7 expression by myeloma cells regulates their pro-angiogenic properties in multiple myeloma patients

Leukemia volume 25, pages 527–537 (2011)Cite this article

Subjects

Abstract

The deregulation of the homeobox genes as homeoboxB (HOXB)-7 has been previously associated to tumor progression and angiogenesis; here we investigated the potential role of HOXB7 in the pro-angiogenic properties of multiple myeloma (MM) cells. We found that HOXB7 was expressed in 10 out of 22 MM patients analyzed at the diagnosis related to high bone marrow angiogenesis and overexpressed in about 40% of myeloma cell lines compared with normal plasma cells. Enforced HOXB7 expression in MM cells by a lentiviral vector significantly modified their transcriptional and angiogenic profile, checked by combined microarray and angiogenesis PCR analyses, upregulating VEGFA, FGF2, MMP2, WNT5a and PDGFA and downregulating thrombospoindin-2. The pro- and anti-angiogenic HOXB7-related gene signature was also validated in a large independent dataset of MM patients. Accordingly, MM-induced vessel formation was significantly increased by HOXB7 overexpression both in vitro angiogenic and chorioallantoic membrane assays, as well as the HOXB7 silencing by small interfering RNA inhibited the production of angiogenic factors, and the pro-angiogenic properties of MM cells. Finally, in SCID-NOD mice we confirmed that HOXB7 overexpression by MM cells stimulated tumor growth, increased MM-associated angiogenesis and the expression of pro-angiogenic genes by microarray analysis supporting the critical role of HOXB7 in the angiogenic switch in MM.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5

Similar content being viewed by others

Accession codes

Accessions

GenBank/EMBL/DDBJ

References

  1. Vacca A, Ribatti D, Presta M, Minischetti M, Iurlaro M, Ria R et al. Bone marrow neovascularization, plasma cell angiogenic potential, and matrix metalloproteinase-2 secretion parallel progression of human multiple myeloma. Blood 1999; 93: 3064–3073.

    CAS  PubMed  Google Scholar 

  2. Vacca A, Ribatti D . Bone marrow angiogenesis in multiple myeloma. Leukemia 2006; 20: 193–199.

    CAS  PubMed  Google Scholar 

  3. Rajkumar SV, Leong T, Roche PC . Prognostic value of bone marrow angiogenesis in multiple myeloma. Clin Cancer Res 2000; 6: 3111–3116.

    CAS  PubMed  Google Scholar 

  4. Rajkumar SV, Mesa RA, Fonseca R, Schroeder G, Plevak MF, Dispenzieri A et al. Bone marrow angiogenesis in 400 patients with monoclonal gammopathy of undetermined significance, multiple myeloma, and primary amyloidosis. Clin Cancer Res 2002; 8: 2210–2216.

    PubMed  Google Scholar 

  5. Munshi NC, Wilson C . Increased bone marrow microvessel density in newly diagnosed multiple myeloma carries a poor prognosis. Semin Oncol 2001; 28: 565–569.

    Article  CAS  PubMed  Google Scholar 

  6. Xu JL, Lai R, Kinoshita T, Nakashima N, Nagasaka T . Proliferation, apoptosis, and intratumoral vascularity in multiple myeloma: correlation with the clinical stage and cytological grade. J Clin Pathol 2002; 55: 530–534.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Alexandrakis MG, Passam FH, Dambaki C, Pappa CA, Stathopoulos EN . The relation between bone marrow angiogenesis and the proliferation index Ki-67 in multiple myeloma. J Clin Pathol 2004; 57: 856–860.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Hose D, Moreaux J, Meissner T, Seckinger A, Goldschmidt H, Benner A et al. Induction of angiogenesis by normal and malignant plasma cells. Blood 2009; 114: 123–148.

    Article  Google Scholar 

  9. Dankbar B, Padro T, Leo R, Feldmann B, Kropff M, Mesters RM et al. Vascular endothelial growth factor and interleukin-6 in paracrine tumor-stromal cell interactions in multiple myeloma. Blood 2000; 95: 2630–2636.

    CAS  PubMed  Google Scholar 

  10. Podar K, Anderson KC . The pathophisiologic role of VEGF in hematologic malignancies: therapeutic implications. Blood 2005; 105: 1383–1395.

    Article  CAS  PubMed  Google Scholar 

  11. Bisping G, Leo R, Wenning D, Dankbar B, Padró T, Kropff M et al. Paracrine interactions of basic fibroblast growth factor and interleukin-6 in multiple myeloma. Blood 2003; 101: 2775–2783.

    Article  CAS  PubMed  Google Scholar 

  12. Shapiro VS, Mollenauer MN, Weiss A . Endogenous CD28 expressed on myeloma cells up-regulates interleukin-8 production: implications for multiple myeloma progression. Blood 2001; 98: 187–193.

    Article  CAS  PubMed  Google Scholar 

  13. Giuliani N, Colla S, Lazzaretti M, Sala R, Roti G, Mancini C et al. Pro-angiogenic properties of human myeloma cells: production of angiopoietin-1 and its potential relationship to myeloma-induced angiogenesis. Blood 2003; 102: 638–645.

    Article  CAS  PubMed  Google Scholar 

  14. Colla S, Morandi F, Lazzaretti M, Rizzato R, Lunghi P, Bonomini S et al. Human myeloma cells express the bone regulating gene Runx2/Cbfa1 and produce osteopontin that is involved in angiogenesis in multiple myeloma patients. Leukemia 2005; 19: 2166–2176.

    Article  CAS  PubMed  Google Scholar 

  15. Van Valckenborgh E, Croucher PI, De Raeve H, Carron C, De Leenheer E, Blacher S et al. Multifunctional role of matrix metalloproteinases in multiple myeloma: a study in the 5T2MM mouse model. Am J Pathol 2004; 165: 869–878.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Barillé S, Akhoundi C, Collette M, Mellerin MP, Rapp MJ, Harousseau JL et al. Metalloproteinases in multiple myeloma: production of matrix metalloproteinase-9 (MMP-9), activation of proMMP-2, and induction of MMP-1 by myeloma cells. Blood 1997; 90: 1649–1655.

    PubMed  Google Scholar 

  17. Gorski DH, Walsh K . The role of homeobox genes in vascular remodeling and angiogenesis. Circ Res 2000; 87: 865–872.

    Article  CAS  PubMed  Google Scholar 

  18. Carè A, Felicetti F, Meccia E, Bottero L, Parenza M, Stoppacciaro A et al. HOXB7: a key factor for tumor-associated angiogenic switch. Cancer Res 2001; 61: 6532–6539.

    PubMed  Google Scholar 

  19. Caré A, Silvani A, Meccia E, Mattia G, Peschle C, Colombo MP . Transduction of the SkBr3 breast carcinoma cell line with the HOXB7 gene induces bFGF expression, increases cell proliferation and reduces growth factor dependence. Oncogene 1998; 16: 3285–3289.

    Article  PubMed  Google Scholar 

  20. Wu X, Chen H, Parker B, Rubin E, Zhu T, Lee JS et al. HOXB7, a homeodomain protein, is overexpressed in breast cancer and confers epithelial-mesenchymal transition. Cancer Res 2006; 66: 9527–9534.

    Article  CAS  PubMed  Google Scholar 

  21. Chen H, Lee JS, Liang X, Zhang H, Zhu T, Zhang Z et al. Hoxb7 inhibits transgenic HER-2/neu-induced mouse mammary tumor onset but promotes progression and lung metastasis. Cancer Res 2008; 68: 3637–3644.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Caré A, Silvani A, Meccia E, Mattia G, Stoppacciaro A, Parmiani G et al. HOXB7 constitutively activates basic fibroblast growth factor in melanomas. Mol Cell Biol 1996; 16: 4842–4851.

    Article  PubMed  PubMed Central  Google Scholar 

  23. Giuliani N, Todoerti K, Lisignoli G, Tagliaferri S, Agnelli L, Morandi F et al. Gene expression profiling of isolated mesenchymal and osteoblastic cells exhibits a different pattern of expression in multiple myeloma patients as compared to healthy subjects: potential relationship with the presence of bone lesions. Blood 2007; 110: 3513a.

    Article  Google Scholar 

  24. Zufferey R, Nagy D, Mendel RJ, Naldini L, Trono D . Multiply attenuated lentiviral vector achieves efficient gene delivery in vivo. Nat Biotechnol 1997; 15: 871–875.

    Article  CAS  PubMed  Google Scholar 

  25. Ribatti D, Nico B, Vacca A, Presta M . The gelatin sponge-chorioallantoic membrane assay. Nat Protoc 2006; 1: 85–91.

    Article  CAS  PubMed  Google Scholar 

  26. Airoldi I, Di Carlo E, Banelli B, Moserle L, Cocco C, Pezzolo A et al. The IL-12Rbeta2 gene functions as a tumor suppressor in human B cell malignancies. J Clin Invest 2004; 113: 1651–1959.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Agnelli L, Mosca L, Fabris S, Lionetti M, Andronache A, Kwee I et al. A SNP microarray and FISH-based procedure to detect allelic imbalances in multiple myeloma: An integrated genomics approach reveals a wide gene dosage effect. Genes Chromosomes Cancer 2009; 48: 603–614.

    Article  CAS  PubMed  Google Scholar 

  28. Lombardi L, Poretti G, Mattioli M, Fabris S, Agnelli L, Bicciato S et al. Molecular characterization of human multiple myeloma cell lines by integrative genomics: insights into the biology of the disease. Genes Chromosomes Cancer 2007; 46: 226–238.

    Article  CAS  PubMed  Google Scholar 

  29. Giampaolo A, Pelosi E, Valtieri M, Montesoro E, Sterpetti P, Samoggia P et al. HOXB gene expression and function in differentiating purified hematopoietic progenitors. Stem Cells 1995; 13 (Suppl 1): 90–105.

    CAS  PubMed  Google Scholar 

  30. Carè A, Valtieri M, Mattia G, Meccia E, Masella B, Luchetti L et al. Enforced expression of HOXB7 promotes hematopoietic stem cell proliferation and myeloid-restricted progenitor differentiation. Oncogene 1999; 18: 1993–2001.

    Article  PubMed  Google Scholar 

  31. Shiraishi K, Yamasaki K, Nanba D, Inoue H, Hanakawa Y, Shirakata Y et al. Pre-B-cell leukemia transcription factor 1 is a major target of promyelocytic leukemia zinc-finger-mediated melanoma cell growth suppression. Oncogene 2007; 26: 339–348.

    Article  CAS  PubMed  Google Scholar 

  32. Yamashita T, Tazawa S, Yawei Z, Katayama H, Kato Y, Nishiwaki K et al. Suppression of invasive characteristics by antisense introduction of overexpressed HOX genes in ovarian cancer cells. Int J Oncol 2006; 28: 931–938.

    CAS  PubMed  Google Scholar 

  33. Raval A, Tanner SM, Byrd JC, Angerman EB, Perko JD, Chen SS et al. Downregulation of death-associated protein kinase 1 (DAPK1) in chronic lymphocytic leukemia. Cell 2007; 129: 879–890.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Novak EM, Linardi C, Metzger M, Maria DA, Giorgi RR, Moura F et al. Differential expression of HOXB7 gene in multiple myeloma and extramedullary multiple myeloma patients. Eur J Haematol 2010; 84: 185–186.

    Article  PubMed  Google Scholar 

  35. Risau W, Drexler H, Mironov V, Smits A, Siegbahn A, Funa K et al. Platelet-derived growth factor is angiogenic in vivo. Growth Factors 1992; 7: 261–266.

    Article  CAS  PubMed  Google Scholar 

  36. de Fraipont F, Nicholson AC, Feige JJ, Van Meir EG . Thrombospondins and tumor angiogenesis. Trends Mol Med 2001; 7: 401–407.

    Article  CAS  PubMed  Google Scholar 

  37. Rendtlew Danielsen JM, Knudsen LM, Dahl IM, Lodahl M, Rasmussen T . Dysregulation of CD47 and the ligands thrombospondin 1 and 2 in multiple myeloma. Br J Haematol 2007; 138: 756–760.

    Article  PubMed  Google Scholar 

  38. Airoldi I, Cocco C, Giuliani N, Ferrarini M, Colla S, Ognio E et al. Constitutive expression of IL-12R beta 2 on human multiple myeloma cells delineates a novel therapeutic target. Blood 2008; 112: 750–759.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Ling L, Nurcombe V, Cool SM . Wnt signaling controls the fate of mesenchymal stem cells. Gene 2009; 433: 1–7.

    Article  CAS  PubMed  Google Scholar 

  40. Masckauchán TN, Agalliu D, Vorontchikhina M . Wnt5a signaling induces proliferation and survival of endothelial cells in vitro and expression of MMP-1 and Tie-2. Mol Biol Cell 2006; 17: 5163–5172.

    Article  PubMed  PubMed Central  Google Scholar 

  41. Huang CL, Liu D, Nakano J, Ishikawa S, Kontani K, Yokomise H et al. Wnt5a expression is associated with the tumor proliferation and the stromal vascular endothelial growth factor—an expression in non-small-cell lung cancer. J Clin Oncol 2005; 23: 8765–8773.

    Article  PubMed  Google Scholar 

  42. O’Connell MP, Fiori JL, Xu M, Carter AD, Frank BP, Camilli TC et al. The orphan tyrosine kinase receptor, ROR2, mediates Wnt5A signaling in metastatic melanoma. Oncogene 2009; 29: 34–44.

    Article  PubMed  PubMed Central  Google Scholar 

  43. Zangari M, Anaissie E, Stopeck A, Morimoto A, Tan N, Lancet J et al. Phase II study of SU5416, a small molecule vascular endothelial growth factor tyrosine kinase receptor inhibitor, in patients with refractory multiple myeloma. Clin Cancer Res 2004; 10: 88–95.

    Article  CAS  PubMed  Google Scholar 

  44. Milanović D, Maier P, Lohr F, Wenz F, Herskind C . Inhibition of 13-cis retinoic acid-induced gene expression of homeobox B7 by thalidomide. Int J Cancer 2007; 121: 1205–1211.

    Article  PubMed  Google Scholar 

Download references

Acknowledgements

Paper was supported by grants from the ‘International Myeloma Foundation’; the ‘Italian Minister of Health-Progetti Regione Emilia Romagna’ and ‘Associazione Italiana per la Ricerca sul Cancro’ A.I.R.C.: IG no 4659 (AN) and IG2009 no 8530 (NG) and the fellowship from ‘Fondazione Italiana Ricerca sul Cancro’ (LA). We thank the ‘Associazione Italiana Contro le Leucemie’ (AIL) Parma section for the financial support.

Author information

Author notes

  1. S Colla

    Present address: 12Current address: Dana Farber Cancer Institute, Boston, MA, USA.,

Authors and Affiliations

  1. Chair of Hematology and BMT Center, University of Parma, Parma, Italy

    P Storti, S Colla, M Bolzoni, M Abeltino, S Bonomini, V Rizzoli & N Giuliani

  2. Dipartimento di Salute Animale’, ‘Sezione Malattie Infettive, University of Parma, Parma, Italy

    G Donofrio & V Franceschi

  3. AIRC Laboratory of Immunology and Tumors, G. Gaslini Institute, Genova, Italy

    I Airoldi

  4. Department of Medical Sciences University of Milan, Hematology 1, Fondazione IRCCS Ca' Granda Ospedale Maggiore, Policlinico, Milano, Italy

    L Agnelli, K Todoerti & A Neri

  5. ‘Dip. Genetica, Biologia dei Microrganismi, Antropologia, Evoluzione’, University of Parma, Parma, Italy

    M Lazzaretti

  6. Pathology, ‘Azienda Ospedaliero-Universitaria di Parma’, Parma, Italy

    C Mancini

  7. Department of Human Anatomy, University of Bari, Italy

    D Ribatti

  8. Department of Experimental and Laboratory Medicine, G. Gaslini Institute, Genova, Italy

    V Pistoia

  9. Laboratorio di Immunologia e Genetica, Istituti Ortopedici Rizzoli, Bologna, Italy

    G Lisignoli

  10. ‘U.O. ‘Patologia dell’Apparato Locomotore’, ‘Azienda Ospedaliero-Universitaria di Parma’, Parma, Italy

    A Pedrazzini

  11. ENT Department, S. Orsola-Malpighi Hospital, Bologna University, Bologna, Italy

    O Cavicchi

Authors
  1. P Storti
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. G Donofrio
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S Colla
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. I Airoldi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. M Bolzoni
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. L Agnelli
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. M Abeltino
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. K Todoerti
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. M Lazzaretti
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. C Mancini
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. D Ribatti
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. S Bonomini
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. V Franceschi
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. V Pistoia
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. G Lisignoli
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. A Pedrazzini
    View author publications

    You can also search for this author in PubMed Google Scholar

  17. O Cavicchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  18. A Neri
    View author publications

    You can also search for this author in PubMed Google Scholar

  19. V Rizzoli
    View author publications

    You can also search for this author in PubMed Google Scholar

  20. N Giuliani
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to N Giuliani.

Ethics declarations

Competing interests

The authors declare no conflict of interests.

Additional information

Supplementary Information accompanies the paper on the Leukemia website

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Storti, P., Donofrio, G., Colla, S. et al. HOXB7 expression by myeloma cells regulates their pro-angiogenic properties in multiple myeloma patients. Leukemia 25, 527–537 (2011). https://doi.org/10.1038/leu.2010.270

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/leu.2010.270

Keywords

This article is cited by

Search

Advanced search

Quick links