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Assessment of minimal residual disease (MRD) in CBFbeta/MYH11-positive acute myeloid leukemias by qualitative and quantitative RT-PCR amplification of fusion transcripts

Leukemia volume 16, pages 1176–1181 (2002)Cite this article

Abstract

The inv(16)(p13q22) chromosomal rearrangement associated with FAB M4Eo acute myeloid leukemia (AML) subtype is characterized by the presence of the CBFbeta/MYH11 fusion transcript that can be used to detect minimal residual disease (MRD). However, qualitative RT-PCR studies of MRD have so far produced conflicting results and seem of limited prognostic value. We have evaluated retrospectively MRD in a large series of CBFbeta/MYH11-positive patients employing both qualitative and quantitative (real-time PCR) approaches. 186 bone marrow samples from 36 patients were examined with a median follow-up of 27.5 months; 15 patients relapsed during follow-up. In qualitative studies, carried out by ‘nested’ RT-PCR assay, all patients in complete remission (CR) immediately after induction/consolidation therapy were found to be PCR positive. However, follow-up samples at later time points were persistently negative (except one case) in patients remaining in continuous CR (CCR) for more than 12 months. 16 patients were evaluated by quantitative real-time PCR assay: CBFbeta/MYH11 transcript copy number was normalized for expression of the housekeeping gene ABL, expressed as fusion gene copy number per 104 copies of ABL. A 2–3 log decline in leukemic transcript copy number was observed after induction/consolidation therapy. After achieving CR, the mean copy number was significantly higher in patients destined to relapse compared to patients remaining in CCR (151 vs 9, P < 0.0001 by Mann–Whitney test). Moreover, in CCR patients, the copy number dropped below the detection threshold after the treatment protocol was completed and remained undetectable in subsequent MRD analysis in accordance with results obtained by qualitative RT-PCR. On the contrary, in the seven patients who relapsed, the copy number in CR never declined below the detection threshold; thus a cut-off value discriminating these two groups of patients could be established. The findings of our study, if confirmed, might confer an important predictive value to quantitative real-time PCR determinations of MRD in patients with inv(16) leukemia.

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Acknowledgements

This work was supported by grants from CNR (Progetto Finalizzato Biotecnologie), MURST-COFIN 2000, AIL (Associazione Italiana contro le Leucemie) and AIRC (Associazione Italiana per la Ricerca sul Cancro). DC is supported by fellowship of the Associazione Italiana Amici di Josè Carreras. We thank the European SANCO Group for useful collaboration in implementing PCR technology. We are indebted to Dr E Aruta for statistical assistance.

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Authors and Affiliations

  1. Dipartimento di Scienze Cliniche e Biologiche, Università di Torino, Orbassano, Torino, Italy

    A Guerrasio, C Pilatrino, D De Micheli, D Cilloni, A Serra, E Gottardi, A Parziale & G Saglio

  2. Divisione di Ematologia, Ospedale San Giovanni Battista, Torino, Italy

    F Marmont

  3. Dipartimento di Biotecnologie Cellulari ed Ematologia, Università ‘La Sapienza’, Roma, Italy

    D Diverio, M Divona & F Lo Coco

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  1. A Guerrasio
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  2. C Pilatrino
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Guerrasio, A., Pilatrino, C., De Micheli, D. et al. Assessment of minimal residual disease (MRD) in CBFbeta/MYH11-positive acute myeloid leukemias by qualitative and quantitative RT-PCR amplification of fusion transcripts. Leukemia 16, 1176–1181 (2002). https://doi.org/10.1038/sj.leu.2402478

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