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Identification of BCR-ABL point mutations conferring resistance to the Abl kinase inhibitor AMN107 (nilotinib) by a random mutagenesis study

Arghya Ray, Sandra W. Cowan-Jacob, Paul W. Manley, Jürgen Mestan and James D. Griffin

Data supplements

Article Figures & Data

Figures

  • Figure 1

    AMN107 treatment of BCR-ABL point mutant and wild-type BCR-ABL–expressing Ba/F3 cell lines. This figure shows the results of AMN107 treatment of different BCR-ABL point mutants and wild-type Bcr-Abl–expressing Ba/F3 cell lines using a wide range of concentrations for 72 hours. The dose response curves obtained from the MTS assay are presented in a semilog plot. Error bars indicate the standard deviation of the mean for each dose.

  • Figure 2

    AMN107 bound to the kinase domain of Abl. The ribbon diagram depicts the kinase domain of Abl (shown in gray) with AMN107 (shown as a transparent surface with its skeleton highlighted) bound within the ATP pocket, and with different point mutants highlighted to show their locations within the kinase domain. The side chains of the mutated residues are color-coded according to the color scheme used in Figure 1, such that E255K is red, Y253H/C is purple, and so on.

Tables

  • Table 1

    Properties of different point mutations

    AMN107-resistant BCR-ABL mutantConcentration at recovery, nMNo. of coloniesPercentage of total*IC50 of AMN107, nMIC50 of imatinib mesylate, nM
    Wild-type< 10450
    K247N12511.1690≈850
    L248V25011.16675≈10 000
    Q252H12522.3240≈1 300
    Y253C25089.30600≈10 000
    Y253H50078.141300≈10 000
    E255K25033.49200≈10 000
    L273F12511.1651750
    E282K25022.32150≈1 550
    K285N25022.32≈500≈3 500
    V289L12511.16251 000
    E292K12511.1690≈1 500
    N297T12511.1630650
    T315I5001112.79≈8000≈10 000
    H375P12511.16501 100
    T406I12511.1680≈1 400
    W430L12522.3290800
    E431G12511.1635≈650
    • Six mutations, namely L248V, Q252H, Y253C, Y253H, E255K, and T315I, identified here from the screen, have previously been reported in patients with CML receiving imatinib mesylate, and are known to be associated with imatinib mesylate resistance. All double-mutant clones were recovered at 500 nM concentration.

    • — indicates no recovery.

    • * (no. of individual colonies/total no. of colonies obtained) × 100.