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:

Myelodisplasias

CDKN2B methylation status and isolated chromosome 7 abnormalities predict responses to treatment with 5-azacytidine

Leukemia volume 21, pages 1937–1944 (2007)Cite this article

Abstract

5-Azacytidine, a DNA methyl transferase inhibitor, is effective in patients with myelodysplastic syndromes (MDS). Whether responses to 5-Azacytidine are achieved by demethylation of key genes or by cytotoxicity is unclear. Of 34 patients with MDS or acute myeloid leukaemia (AML) treated with 5-Azacytidine, 7 achieved complete remissions (CR) (21%) and 6 achieved haematological improvement. All six had less than 5% bone marrow (BM) blasts at the time of haematological improvements (HI) (2 had pre-existing refractory anaemia (RA), 4 had refractory anaemia with excess blasts (RAEB)). A further patient with RAEB had blast reduction to less than 5% without HI. Five of the seven (71%) complete responders had chromosome 7 abnormalities. BM CR predicted longer overall survival (OS) (median 23 versus 9 months, P=0.015). Bisulphite genomic sequencing (BGS) of the CDKN2B (p15INK4b) promoter showed low level, heterogeneous pretreatment methylation (mean 12.2%) in 14/17 (82%) patients analysed. Lower baseline methylation occurred in responders (9.8% versus 16.2% in non-responders P=0.07). No response was seen in patients with >24% methylation, in whom p15INK4b mRNA was not expressed. 5-Azacytidine reduced CDKN2B methylation by mean 6.8% in 8/17 (47%) patients, but this did not correlate with response. At 75 mg/m2, cell death (reduced BM cellularity (P=0.001) and increased apoptosis (P=0.02)) rather than demethylation of CDKN2B correlates with response. Patients with >24% methylation may benefit from alternative dosing or combination strategies.

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

Similar content being viewed by others

References

  1. Cheson BD, Bennett JM, Kantarjian H, Pinto A, Schiffer CA, Nimer SD et al. Report of an international working group to standa. Blood 2000; 96: 3671–3674.

    CAS  PubMed  Google Scholar 

  2. Greenberg P, Cox C, LeBeau MM, Fenaux P, Morel P, Sanz G et al. International scoring system for evaluating prognosis in myelodysplastic syndromes. Blood 1997; 89: 2079–2088.

    CAS  PubMed  Google Scholar 

  3. Germing U, Hildebrandt B, Pfeilstocker M, Nosslinger T, Valent P, Fonatsch C et al. Refinement of the international prognostic scoring system (IPSS) by including LDH as an additional prognostic variable to improve risk assessment in patients with primary myelodysplastic syndromes (MDS). Leukemia 2005; 19: 2223–2231.

    Article  CAS  PubMed  Google Scholar 

  4. Fukumoto JS, Greenberg PL . Management of patients with higher risk myelodysplastic syndromes. Crit Rev Oncol/Hematol Acute Leukemias 2005; 56: 179–192.

    Article  Google Scholar 

  5. Teofili L, Morosetti R, Martini M, Urbano R, Putzulu R, Rutella S et al. Expression of cyclin-dependent kinase inhibitor p15(INK4B) during normal and leukemic myeloid differentiation. Exp Hematol 2000; 28: 519–526.

    Article  CAS  PubMed  Google Scholar 

  6. Teofili L, Martini M, Di Mario A, Rutella S, Urbano R, Luongo M et al. Expression of p15(ink4b) gene during megakaryocytic differentiation of normal and myelodysplastic hematopoietic progenitors. Blood 2001; 98: 495–497.

    Article  CAS  PubMed  Google Scholar 

  7. Quesnel B, Guillerm G, Vereecque R, Wattel E, Preudhomme C, Bauters F et al. Methylation of the p15(INK4b) gene in myelodysplastic syndromes is frequent and acquired during disease progression. Blood 1998; 91: 2985–2990.

    CAS  PubMed  Google Scholar 

  8. Tien HF, Tang JH, Tsay W, Liu MC, Lee FY, Wang CH et al. Methylation of the p15(INK4B) gene in myelodysplastic syndrome: it can be detected early at diagnosis or during disease progression and is highly associated with leukaemic transformation. Br J Haematol 2001; 112: 148–154.

    Article  CAS  PubMed  Google Scholar 

  9. Aggerholm A, Guldberg P, Hokland M, Hokland P . Extensive intra- and interindividual heterogeneity of p15INK4B methylation in acute myeloid leukemia. Cancer Res 1999; 59: 436–441.

    CAS  PubMed  Google Scholar 

  10. Chen W, Zhang W, Zhu J . Homozygous deletion and methylation of p16 and p15 gene in acute leukemia]. Zhonghua Xue Ye Xue Za Zhi 1999; 20: 474–476.

    CAS  PubMed  Google Scholar 

  11. Christiansen DH, Andersen MK, Pedersen-Bjergaard J . Methylation of p15INK4B is common, is associated with deletion of genes on chromosome arm 7q and predicts a poor prognosis in therapy-related myelodysplasia and acute myeloid leukemia. Leukemia 2003; 17: 1813–1819.

    Article  CAS  PubMed  Google Scholar 

  12. Christman JK . 5-Azacytidine and 5-aza-2′-deoxycytidine as inhibitors of DNA methylation: mechanistic studies and their implications for cancer therapy. Oncogene 2002; 21: 5483–5495.

    Article  CAS  PubMed  Google Scholar 

  13. Silverman LR, Demakos EP, Peterson BL, Kornblith AB, Holland JC, Odchimar-Reissig R et al. Randomized controlled trial of azacitidine in patients with the myelodysplastic syndrome: a study of the cancer and leukemia group B. J Clin Oncol 2002; 20: 2429–2440.

    Article  CAS  PubMed  Google Scholar 

  14. Cheson BD, Greenberg PL, Bennett JM, Lowenberg B, Wijermans PW, Nimer SD et al. Clinical application and proposal for modification of the International Working Group (IWG) response criteria in myelodysplasia. Blood 2006; 108: 419–425.

    Article  CAS  PubMed  Google Scholar 

  15. Lea NC, Orr SJ, Stoeber K, Williams GH, Lam EW-F, Ibrahim MAA, Mufti GJ, Thomas NSB . Commitment point during G0 → G1 that controls entry into the cell cycle. Mol Cell Biol 2003; 23: 2351–2361.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Frommer M, McDonald L, Millar D, Collis C, Watt F, Grigg G et al. A genomic sequencing protocol that yields a positive display of 5-methylcytosine Residues in Individual DNA Strands. Proc Natl Acad Sci USA 1992; 89: 1827–1831.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Cameron EE, Baylin SB, Herman JG . p15(INK4B) CpG island methylation in primary acute leukemia is heterogeneous and suggests density as a critical factor for transcriptional silencing. Blood 1999; 94: 2445–2451.

    CAS  PubMed  Google Scholar 

  18. Koopman G, Reutelingsperger C, Kuijten G, Keehnen R, Pals S, van Oers M . Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis. Blood 1994; 84: 1415–1420.

    CAS  PubMed  Google Scholar 

  19. Mitelman F . ISCN Guidelines for Cancer Cytogenetics, supplement to and International System for Human Cytogenetic Nomenclature. Karger: Basel, Switzerland, 1991.

    Google Scholar 

  20. Herman JG, Graff JR, Myohanen S, Nelkin BD, Baylin SB . Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci USA 1996; 93: 9821–9826.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Lubbert M, Wijermans P, Kunzmann R, Verhoef G, Bosly A, Ravoet C et al. Cytogenetic responses in high-risk myelodysplastic syndrome following low-dose treatment with the DNA methylation inhibitor 5-aza-2′-deoxycytidine. Br J Haematol 2001; 114: 349–357.

    Article  CAS  PubMed  Google Scholar 

  22. Hagop Kantarjian J-PJI, Rosenfeld CS, Bennett JM, Albitar M, DiPersio J, Klimek V et al. Decitabine improves patient outcomes in myelodysplastic syndromes. Cancer 2006; 106: 1794–1803.

    Article  PubMed  Google Scholar 

  23. Dodge JE, List AF, Futscher BW . Selective variegated methylation of the p15 CpG island in acute myeloid leukemia. Int J Cancer 1998; 78: 561–567.

    Article  CAS  PubMed  Google Scholar 

  24. Tessema M, Langer F, Dingemann J, Ganser A, Kreipe H, Lehmann U . Aberrant methylation and impaired expression of the p15(INK4b) cell cycle regulatory gene in chronic myelomonocytic leukemia (CMML). Leukemia 2003; 17: 910–918.

    Article  CAS  PubMed  Google Scholar 

  25. Mufti GJ, Figes A, Hamblin TJ, Oscier DG, Copplestone JA . Immunological abnormalities in myelodysplastic syndromes. I. Serum immunoglobulins and autoantibodies. Br J Haematol 1986; 63: 143–147.

    Article  CAS  PubMed  Google Scholar 

  26. Copplestone JA, Mufti GJ, Hamblin TJ, Oscier DG . Immunological abnormalities in myelodysplastic syndromes. II. Coexistent lymphoid or plasma cell neoplasms: a report of 20 cases unrelated to chemotherapy. Br J Haematol 1986; 63: 149–159.

    Article  CAS  PubMed  Google Scholar 

  27. Wlodarski MW, Gondek LP, Nearman ZP, Plasilova M, Kalaycio M, Hsi ED et al. Molecular strategies for detection and quantitation of clonal cytotoxic T-cell responses in aplastic anemia and myelodysplastic syndrome. Blood 2006; 108: 2632–2641.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Yang AS, Doshi KD, Choi SW, Mason JB, Mannari RK, Gharybian V et al. DNA methylation changes after 5-aza-2′-deoxycytidine therapy in patients with leukemia. Cancer Res 2006; 66: 5495–5503.

    Article  CAS  PubMed  Google Scholar 

  29. Kantarjian H, Oki Y, Garcia-Manero G, Huang X, O'Brien S, Cortes J et al. Results of a randomized study of 3 schedules of low-dose decitabine in higher-risk myelodysplastic syndrome and chronic myelomonocytic leukemia. Blood 2007; 109: 52–57.

    Article  CAS  PubMed  Google Scholar 

  30. Garcia-Manero G, Kantarjian HM, Sanchez-Gonzalez B, Yang H, Rosner G, Verstovsek S et al. Phase 1/2 study of the combination of 5-aza-2′-deoxycytidine with valproic acid in patients with leukemia. Blood 2006; 108: 3271–3279.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Daskalakis M, Nguyen TT, Nguyen C, Guldberg P, Kohler G, Wijermans P et al. Demethylation of a hypermethylated P15/INK4B gene in patients with myelodysplastic syndrome by 5-Aza-2′-deoxycytidine (decitabine) treatment. Blood 2002; 100: 2957–2964.

    Article  CAS  PubMed  Google Scholar 

  32. Gore SD, Baylin S, Sugar E, Carraway H, Miller CB, Carducci M et al. Combined DNA methyltransferase and histone deacetylase inhibition in the treatment of myeloid neoplasms. Cancer Res 2006; 66: 6361–6369.

    Article  CAS  PubMed  Google Scholar 

  33. Stresemann C, Brueckner B, Musch T, Stopper H, Lyko F . Functional diversity of DNA methyltransferase inhibitors in human cancer cell lines. Cancer Res 2006; 66: 2794–2800.

    Article  CAS  PubMed  Google Scholar 

  34. Parker JE, Mufti GJ, Rasool F, Mijovic A, Devereux S, Pagliuca A . The role of apoptosis, proliferation, and the Bcl-2-related proteins in the myelodysplastic syndromes and acute myeloid leukemia secondary to MDS. Blood 2000; 96: 3932–3938.

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

Kavita Raj was funded by the Medical Research Council, UK. Alison John and Constantinos Chronis were funded by the Leukaemia Research Fund.

Author information

Authors and Affiliations

  1. Department of Haematological Medicine, King's College London, Denmark Hill Campus, London, UK

    K Raj, A John, A Ho, C Chronis, S Khan, J Samuel, N S B Thomas & G J Mufti

  2. Department of Histopathology, King's College London, Denmark Hill Campus, London, UK

    S Pomplun

Authors
  1. K Raj
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A John
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. A Ho
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. C Chronis
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. S Khan
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. J Samuel
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. S Pomplun
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. N S B Thomas
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. G J Mufti
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to G J Mufti.

Additional information

Supplementary Information accompanies the paper on the Leukemia website (http://www.nature.com/leu)

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Raj, K., John, A., Ho, A. et al. CDKN2B methylation status and isolated chromosome 7 abnormalities predict responses to treatment with 5-azacytidine. Leukemia 21, 1937–1944 (2007). https://doi.org/10.1038/sj.leu.2404796

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.leu.2404796

Keywords

This article is cited by

Search

Advanced search

Quick links