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Acute Leukemias

Mutations of FLT3/ITD confer resistance to multiple tyrosine kinase inhibitors

Leukemia volume 27, pages 48–55 (2013)Cite this article

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Abstract

FMS-like tyrosine kinase 3 (FLT3) normally functions in the survival/proliferation of hematopoietic stem/progenitor cells, but its constitutive activation by internal tandem duplication (ITD) mutations correlates with a poor prognosis in AML. The development of FLT3 tyrosine kinase inhibitors (TKI) is a promising strategy, but resistance that arises during the course of treatment caused by secondary mutations within the mutated gene itself poses a significant challenge. In an effort to predict FLT3 resistance mutations that might develop in patients, we used saturation mutagenesis of FLT3/ITD followed by selection of transfected cells in FLT3 TKI. We identified F621L, A627P, F691L and Y842C mutations in FLT3/ITD that confer varying levels of resistance to FLT3 TKI. Western blotting confirmed that some FLT3 TKI were ineffective at inhibiting FLT3 autophosphorylation and signaling through MAP kinase, STAT5 and AKT in some mutants. Balb/c mice transplanted with the FLT3/ITD Y842C mutation confirmed resistance to sorafenib in vivo but not to lestaurtinib. These results indicate a growing number of FLT3 mutations that are likely to be encountered in patients. Such knowledge, combined with known remaining sensitivity to other FLT3 TKI, will be important to establish as secondary drug treatments that can be substituted when these mutants are encountered.

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Acknowledgements

We are grateful to Dr Linzhao Cheng (Johns Hopkins University) for providing us with the L3GFP vector used to visualize FLT3/ITD engraftment and to members of the lab for numerous thoughtful discussions. This work was supported by grants from the NCI (CA90770 and CA90668), Leukemia and Lymphoma Society and Giant Food Pediatric Cancer Research Fund. DS is also supported by the Kyle Haydock Professorship.

Author contributions

ABW designed experiments, performed research, analyzed data and wrote the manuscript; LL and BN performed research; ML analyzed data; PB and DL analyzed data and wrote the manuscript; DS designed experiments, supervised the project, analyzed data and wrote the manuscript.

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Authors and Affiliations

  1. Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    A B Williams, B Nguyen, L Li, P Brown, M Levis & D Small

  2. Department of Biophysics, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    D Leahy

  3. Department of Pediatrics, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    P Brown & D Small

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  1. A B Williams
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Correspondence to D Small.

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Williams, A., Nguyen, B., Li, L. et al. Mutations of FLT3/ITD confer resistance to multiple tyrosine kinase inhibitors. Leukemia 27, 48–55 (2013). https://doi.org/10.1038/leu.2012.191

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