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Sequencing of subcloned PCR products facilitates earlier detection of BCR-ABL1T315I and other mutants compared to direct sequencing of the ABL1 kinase domain

Leukemia volume 22, pages 885–888 (2008)Cite this article

Single-point mutations within the kinase domain (KD) of BCR-ABL1 constitute the most frequent cause of acquired resistance to tyrosine kinase inhibitor (TKI) therapy in patients with chronic myeloid leukemia (CML). Particularly, worrisome is the development of the BCR-ABL1T315I mutation, which affects a highly conserved ‘gatekeeper’ threonine residue near the ABL catalytic domain, which causes steric hindrance that precludes the binding of TKIs to the ABL kinase.1 As a result, BCR-ABL1T315I confers resistance to imatinib and second generation inhibitors (dasatinib and nilotinib), retaining kinase activity even in the presence of micromolar concentrations of these compounds.2 Since the BCR-ABL1T315I mutation has been found to precede or accompany the progression to advanced-phase CML and anticipates failure to TKI therapy,3 early detection of this mutant is important both from a prognostic and a therapeutic standpoint.

Current guidelines recommend KD mutation screening in patients with advanced-phase disease and in those in chronic phase (CP) who exhibit inadequate initial response or signs of loss of response.4 At present, direct sequencing (DS) of nested PCR-amplified BCR-ABL1 products is the favored method for routine clinical use.3 However, DS fails to reliably detect mutant clones representing 20% or less of the BCR-ABL-expressing cells.3 More sensitive methods are therefore necessary to detect the incipient expansion of leukemic clones harboring BCR-ABL1T315I. One such assay involves subcloning and DNA expansion of specific clones (DESC) followed by DNA sequencing.

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Authors and Affiliations

  1. Department of Leukemia, MD Anderson Cancer Center, Houston, TX, USA

    A Quintás-Cardama, H Kantarjian & J Cortes

  2. Division of Cancer Medicine, MD Anderson Cancer Center, Houston, TX, USA

    D L Gibbons

  3. Division of Hematology/Oncology, University of Michigan Comprehensive Cancer Center, Ann Arbor, MI, USA

    M Talpaz & N Donato

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  1. A Quintás-Cardama
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  2. D L Gibbons
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  3. H Kantarjian
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  6. J Cortes
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Correspondence to A Quintás-Cardama.

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Quintás-Cardama, A., Gibbons, D., Kantarjian, H. et al. Sequencing of subcloned PCR products facilitates earlier detection of BCR-ABL1T315I and other mutants compared to direct sequencing of the ABL1 kinase domain. Leukemia 22, 885–888 (2008). https://doi.org/10.1038/leu.2008.42

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