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Selecting and deselecting imatinib-resistant clones: observations made by longitudinal, quantitative monitoring of mutated BCR-ABL

Leukemia volume 19, pages 2159–2165 (2005)Cite this article

Abstract

Resistance to imatinib during the treatment of chronic myeloid leukaemia (CML) is frequently associated with point mutations in the ABL gene encoding the ATP binding region likely to cause disease relapse. Early diagnosis and monitoring of these mutations may be important in order to prevent rapid expansion of resistant clones. We describe a quantitative mutation-specific PCR assay based on the readily available Taqman platform. Selectivity for the mutated target is conferred by mutation-specific primers destabilised by additional mismatches. The assay can be carried out in parallel to standard BCR-ABL quantification and is therefore more quickly compared to standard sequencing procedures. The sensitivity of the assay reaches 0.1%. It also allows for quantitative assessment of mutated clones. By analysing sequential samples of resistant subjects, we show how mutated clones were selected, maintained or deselected depending on the individual treatment setting. The high sensitivity and practical merits of this method makes it a good candidate for prospective molecular surveillance of patients at high risk for imatinib resistance.

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Acknowledgements

This work was supported by scientific grants from the Northern Norwegian Health Authorities and the Norwegian Cancer Association. We thank Geir Bjørkøy, Terje Johansen and Ole Peter Rekvig for critical discussion of the project, Per Arne Standal, Malvin Sjo, Alf K Haaland, Aleksandra Silje and Nina Gulbrandsen for sending patient samples and Labforum for sequencing the samples. Special thanks to Kimmo Porkka for discussing molecular features of CML and Rasmus Goll for help with statistics.

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Authors and Affiliations

  1. Department of Immunology and Transfusion Medicine, University Hospital of Northern Norway, University of Tromsø, Tromsø, Norway

    F X E Gruber, M Olsen & B Skogen

  2. Biochemistry Department, Institute of Medical Biology, University of Tromsø, Tromsø, Norway

    T Lamark & M A Sovershaev

  3. Department of Pharmacology, Institute of Pharmacy, University of Tromsø, Tromsø, Norway

    A Ånonli

  4. Medical Department, Section of Hematology, Rikshospitalet University Hospital, Oslo, Norway

    T Gedde-Dahl

  5. Medical Department, Section of Hematology, St Olavs University Hospital, Trondheim, Norway

    H Hjort-Hansen

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  1. F X E Gruber
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  2. T Lamark
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Correspondence to F X E Gruber.

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

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Gruber, F., Lamark, T., Ånonli, A. et al. Selecting and deselecting imatinib-resistant clones: observations made by longitudinal, quantitative monitoring of mutated BCR-ABL. Leukemia 19, 2159–2165 (2005). https://doi.org/10.1038/sj.leu.2403983

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