Tyrosine kinase inhibitor resistance in chronic myeloid leukemia cell lines: investigating resistance pathways

Abstract

There are three currently identified secondary resistance mechanisms observed in patients with chronic myeloid leukemia (CML) receiving tyrosine kinase inhibitors (TKIs). These are BCR–ABL kinase domain (KD) mutations, increased BCR–ABL expression, and overexpression of drug-efflux proteins (ABCB1 and ABCG2). To investigate the interplay between these three modes of resistance, three CML blast crisis cell lines (K562, its ABCB1-overexpressing variant K562 Dox, and KU812) were cultured in gradually increasing concentrations of imatinib to 2 μM, or dasatinib to 200 nM. Eight imatinib- and two dasatinib-resistant cell lines were established. Two imatinib-resistant K562 lines both had increased BCR–ABL expression as the apparent mode of resistance. However, when a dasatinib-resistant K562 culture was generated we observed gradually increasing BCR–ABL expression which peaked prior to identification of the T315I mutation. BCR–ABL overexpression followed by mutation development was observed in a further 4/10 cell lines, each with different KD mutations. In contrast, three imatinib-resistant K562 Dox lines exhibited only a further increase in ABCB1 expression. All TKI-resistant cell lines generated had increased IC₅₀ (dose of drug required to reduce phosphorylation of the adaptor protein p-Crkl by 50%) to imatinib, dasatinib, and nilotinib, regardless of which TKI was used to induce resistance. This suggests that currently available TKIs share the same susceptibilities to drug resistance.

Keywords::

• Drug resistance
• myeloid leukemias and dysplasias
• cell lines and animal models

Acknowledgements

Nilotinib (Tasigna) and imatinib mesylate (Glivec) were kindly provided by Novartis Pharmaceuticals, Basel, Switzerland. Dasatinib (Sprycel) was kindly provided by Bristol-Myers Squibb, Victoria, Australia. A/Prof. Deborah White receives Honoraria and Research Funding from Novartis Pharmaceuticals and Bristol-Myers Squibb. Prof. Timothy Hughes receives Honoraria and Research Funding, and is on the Speakers Bureau, for Novartis Pharmaceuticals and Bristol-Myers Squibb.

Potential conflict of interest:

Disclosure forms provided by the authors are available with the full text of this article at www.informahealthcare.com/lal.