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 Manuscript
  • Published:

Minimal Residual Disease (MRD)

Quantification of CBFβ/MYH11 fusion transcript by Real Time RT-PCR in patients with INV(16) acute myeloid leukemia

Leukemia volume 15, pages 1072–1080 (2001)Cite this article

Abstract

Amplification of the CBFβ/MYH11 fusion transcript by a qualitative reverse transcription-polymerase chain reaction (RT-PCR) has been used to detect minimal residual disease (MRD) and assess the risk for disease relapse in inv(16)(p13q22) acute myeloid leukemia (AML). This strategy has, however, produced conflicting results and because of an uncertain predictive value, its use in the clinical setting cannot be recommended. The objective of the current study was to evaluate if quantification by Real Time RT-PCR could be useful to determine levels of CBFβ/MYH11 fusion transcripts predictive of clinical outcome in inv(16)(p13q22) AML at diagnosis or during remission. Bone marrow (BM) samples from 16 patients with inv(16) AML enrolled on a German multicenter trial (AML HD93) were analyzed for levels of CBFβ/MYH11 fusion transcripts by Real Time RT-PCR at diagnosis (n = 14), during remission (n = 10) and at relapse (n = 6). The CBFβ/MYH11 transcript copy number in each sample was normalized to copies of an internal control housekeeping transcript (ie 18S). The copy number measured at diagnosis or relapse were 3 to 4 log higher that those measured during remission, following completion of induction treatment. A high CBFβ/MYH11transcript copy number at diagnosis had a significant correlation with a high percentage of BM blasts (Spearman's coefficient = −0.66; P = 0.03), and a borderline correlation with a short complete remission (CR) duration (Spearman's coefficient = −0.51; P = 0.07). No difference in levels of CBFβ/MYH11 fusion transcripts measured during intensification therapy was found between patients destined to relapse and those who continued in CCR (P = 0.75). Following completion of the entire chemotherapy program, patients that during CR showed a CBFβ/MYH11 fusion transcript copy number >10 had a significantly shorter CR duration (P = 0.002) and higher risk for disease relapse (P = 0.05) than patients with a CBFβ/MYH11fusion transcript copy number <10. The results of the current study, therefore, suggest that it is possible to determine in remission samples a threshold of CBFβ/MYH11 transcript copy number above which relapse occurs and below which continuous CR is likely.

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

References

  1. Mrózek K, Bloomfield CD . Chromosome aberrations in de novo acute myeloid leukemia in adults: clinical implications Rev Clin Exp Hematol 1998 5: 44–67

    Google Scholar 

  2. Bloomfield CD, Lawrence D, Byrd JC, Carroll A, Pettenati MJ, Tantravahi R, Patil SR, Davey FR, Berg DT, Schiffer CA, Arthur DC, Mayer RJ . Frequency of prolonged remission duration following high-dose cytarabine intensification in acute myeloid leukemia varies by cytogenetic subtype Cancer Res 1998 58: 4173–4179

    CAS  Google Scholar 

  3. Yin JA, Tobal K . Detection of minimal residual disease in acute myeloid leukaemia: methodologies, clinical and biological significance Br J Haematol 1999 106: 578–590

    Article  CAS  Google Scholar 

  4. Liu P, Tarle SA, Hajra A, Claxton DF, Marlton P, Freedman M, Siciliano MJ, Collins FS . Fusion between transcription factor CBFβ/PEBP2b and a myosin heavy chain in acute myeloid leukemia Science 1993 261: 1041–1044

    Article  CAS  Google Scholar 

  5. Liu PP, Hajra A, Wijmenga C, Collins FS . Molecular pathogenesis of the chromosome 16 inversion in the M4Eo subtype of acute myeloid leukemia Blood 1995 85: 2289–2302

    CAS  PubMed  Google Scholar 

  6. Claxton DF, Liu P, Hsu HB, Marlton P, Hester J, Collins F, Deisseroth AB, Rowley JD, Siciliano MJ . Detection of fusion transcripts generated by the inversion 16 chromosome in acute myelogenous leukemia Blood 1994 83: 1750–1756

    CAS  PubMed  Google Scholar 

  7. Hebert J, Cayuela JM, Daniel MT, Berger R, Sigaux F . Detection of minimal residual disease in acute myelomonocytic leukemia with abnormal marrow eosinophils by nested polymerase chain reaction with allele specific amplification Blood 1994 84: 2291–2296

    CAS  PubMed  Google Scholar 

  8. Tobal K, Johnson PR, Saunders MJ, Harrison CJ, Liu Yin JA . Detection of CBFβ/MYH11 transcripts in patients with inversion and other abnormalities of chromosome 16 at presentation and remission Br J Haematol 1995 91: 104–108

    Article  CAS  Google Scholar 

  9. Poirel H, Radford-Weiss I, Rack K, Troussard X, Veil A, Valensi F, Picard F, Guesnu M, Leboeuf D, Melle J . Detection of the chromosome 16 CBF beta-MYH11 fusion transcript in myelomonocytic leukemias Blood 1995 85: 1313–1322

    CAS  Google Scholar 

  10. Elmaagacli AH, Beelen DW, Kroll M, Trzensky S, Stein C, Schaefer UW . Detection of CBFbeta/MYH11 fusion transcript in patients with inv(16) acute myeloid leukemia after allogeneic bone marrow or peripheral blood progenitor cell transplantation Bone Marrow Transplant 1998 21: 159–166

    Article  CAS  Google Scholar 

  11. Costello R, Sainty D, Blaise D, Gastaut JA, Gabert J, Poirel H, Buzyn-Veil A, Macintyre E . Prognosis value of residual disease monitoring by polymerase chain reaction in patients with CBFbeta/MYH11-positive acute myeloblastic leukemia Blood 1997 89: 2222–2223

    CAS  PubMed  Google Scholar 

  12. Marcucci G, Caligiuri MA, Bloomfield CD . Defining the ‘absence’ of the CBFβ/MYH11 fusion transcript in patients with acute myeloid leukemia and inversion of chromosome 16 to predict long-term complete remission: a call for definitions Blood 1997 90: 5022–5024

    CAS  PubMed  Google Scholar 

  13. Evans PA, Short MA, Jack AS, Norfolk DR, Child JA, Shiach CR, Davies F, Tobal K, Liu Yin JA, Morgan GJ . Detection and quantitation of the CBFbeta/MYH11 transcripts associated with the inv(16) in presentation and follow-up samples from patients with AML Leukemia 1997 11: 364–369

    Article  CAS  Google Scholar 

  14. Laczika K, Novak M, Hilgarth B, Mitterbauer M, Mitterbauer G, Scheidel-Petrovic A, Scholten C, Thalhammer-Scherrer R, Brugger S, Keil F, Schwarzinger I, Haas OA, Lechner K, Jaeger U . Competitive CBFbeta/MYH11 reverse-transcription polymerase chain reaction for quantitative assessment of minimal residual disease during postremission therapy in acute myeloid leukemia with inversion(16): a pilot study J Clin Oncol 1998 16: 1519–1525

    Article  CAS  Google Scholar 

  15. Marcucci G, Livak KJ, Bi WL, Strout MP, Bloomfield CD, Caligiuri MA . Detection of the AML1/ETO fusion transcript in patients with t(8;21)-associated AML using a novel ‘real time’ quantitative RT-PCR assay Leukemia 1998 12: 1482–1489

    Article  CAS  Google Scholar 

  16. Wattjes MP, Krauter J, Nagel S, Heidenreich O, Ganser A, Heil G . Comparison of nested competitive RT-PCR and real-time RT-PCR for the detection and quantification of AML1/MTG8 fusion transcripts in t(8;21) positive acute myelogenous leukemia Leukemia 2000 14: 329–335

    Article  CAS  Google Scholar 

  17. Heid CA, Stevens J, Livak KJ, Williams PM . Real time quantitative PCR Genome Res 1996 6: 986–994

    Article  CAS  Google Scholar 

  18. Tobal K, Newton J, Macheta M, Chang J, Morgenstern G, Evans PA, Morgan G, Lucas GS, Liu Yin JA . Molecular quantitation of minimal residual disease in acute myeloid leukemia with t(8;21) can identify patients in durable remission and predict clinical relapse Blood 2000 95: 815–819

    CAS  PubMed  Google Scholar 

  19. Castilla LH, Garrett L, Adya N, Orlic D, Dutra A, Anderson S, Owens J, Eckhaus M, Bodine D, Liu PP . The fusion gene Cbfb-MYH11 blocks myeloid differentiation and predisposes mice to acute myelomonocytic leukaemia Nat Genet 1999 2: 144–146

    Article  Google Scholar 

  20. Golub TR, Slonim DK, Tamayo P, Huard C, Gaasenbeek M, Mesirov JP, Coller H, Loh ML, Downing JR, Caligiuri MA, Bloomfield CD, Lander ES . Molecular classification of cancer: class discovery and class prediction by gene expression monitoring Science 1999 286: 531–537

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported in part by grant P30CA16058, National Cancer Institute, Bethesda, MD, The Coleman Leukemia Research Foundation and The American Cancer Society IRG-98–278–01 grant.

Author information

Authors and Affiliations

  1. Division of Hematology-Oncology, Department of Internal Medicine, Comprehensive Cancer Center, The Ohio State University, Columbus, OH, USA

    G Marcucci, MA Caligiuri, KJ Archer, EA Maghraby & CD Bloomfield

  2. Division of Hematology-Oncology, University of Ulm, Germany

    H Döhner, RF Schlenk & K Döhner

Authors
  1. G Marcucci
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. MA Caligiuri
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. H Döhner
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. KJ Archer
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. RF Schlenk
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. K Döhner
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. EA Maghraby
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. CD Bloomfield
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Marcucci, G., Caligiuri, M., Döhner, H. et al. Quantification of CBFβ/MYH11 fusion transcript by Real Time RT-PCR in patients with INV(16) acute myeloid leukemia. Leukemia 15, 1072–1080 (2001). https://doi.org/10.1038/sj.leu.2402159

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.leu.2402159

Keywords

This article is cited by

Search

Advanced search

Quick links