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Sensitivity and Resistance to Therapy

DNA repair contributes to the drug-resistant phenotype of primary acute myeloid leukaemia cells with FLT3 internal tandem duplications and is reversed by the FLT3 inhibitor PKC412

Leukemia volume 20, pages 2130–2136 (2006)Cite this article

Abstract

The presence of internal tandem duplications (ITD) mutations in the FMS-like tyrosine kinase 3 (FLT3) receptor influences the risk of relapse in acute myeloid leukaemia (AML). We have investigated DNA repair in FLT3-ITD and wild-type (WT) cells. Using the comet assay, we have demonstrated that the FLT3 inhibitor PKC412 significantly inhibits repair of DNA damage in the MV4-11-FLT3-ITD cell line and FLT3-ITD patient samples but not in the HL-60-FLT3-WT cell line or FLT3-WT patient samples. Following the discovery that transcript levels of the DNA repair gene RAD51 are significantly correlated with FLT3 transcript levels in FLT3-ITD patients, we further investigated the role of RAD51 in FLT3-ITD-AML. The reduction in DNA repair in PKC412-treated FLT3-ITD cells was shown to be associated with downregulation of RAD51 mRNA and protein expression and correlates with the maintenance of phosphorylated H2AX levels, implying that PKC412 inhibits the homologous recombination double-strand break repair pathway in FLT3-ITD cells. Using FLT3-short interfering RNA (siRNA), we also demonstrated that genetic silencing of FLT3 results in RAD51 downregulation in FLT3-ITD cells but not in FLT3-WT cells. This work suggests that the use of FLT3 inhibitors such as PKC412 may reverse the drug-resistant phenotype of FLT3-ITD-AML cells by inhibiting repair of chemotherapy-induced genotoxic damage and thereby reduce the risk of disease relapse.

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Acknowledgements

We acknowledge the NCRI adult working party for permitting access to stored samples from AML trial patients and thank Novartis for providing PKC412. This work was funded by the Leukaemia Research Fund, UK.

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Authors and Affiliations

  1. Department of Academic Haematology, School of Molecular Medical Sciences, Nottingham University Hospitals and University of Nottingham, Nottingham, UK

    C H Seedhouse, H M Hunter, B Lloyd-Lewis, A-M Massip, M Pallis, G I Carter, M Grundy, S Shang & N H Russell

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  1. C H Seedhouse
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  2. H M Hunter
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Correspondence to C H Seedhouse.

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Seedhouse, C., Hunter, H., Lloyd-Lewis, B. et al. DNA repair contributes to the drug-resistant phenotype of primary acute myeloid leukaemia cells with FLT3 internal tandem duplications and is reversed by the FLT3 inhibitor PKC412. Leukemia 20, 2130–2136 (2006). https://doi.org/10.1038/sj.leu.2404439

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