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Internal tandem duplication of FLT3 deregulates proliferation and differentiation and confers resistance to the FLT3 inhibitor AC220 by Up-regulating RUNX1 expression in hematopoietic cells

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Abstract

Internal tandem duplication in the FLT3 gene (FLT3/ITD), which is found in patients with acute myeloid leukemia (AML), causes resistance to FLT3 inhibitors. We found that RUNX1, a transcription factor that regulates normal hematopoiesis, is up-regulated in patients with FLT3/ITD+ AML. While RUNX1 can function as a tumor suppressor, recent data have shown that RUNX1 is required for AML cell survival. In the present study, we investigated the functional role of RUNX1 in FLT3/ITD signaling. FLT3/ITD induced growth factor-independent proliferation and impaired G-CSF mediated myeloid differentiation in 32D hematopoietic cells, coincident with up-regulation of RUNX1 expression. Silencing of RUNX1 expression significantly decreased proliferation and secondary colony formation, and partially abrogated the impaired myeloid differentiation of FLT3/ITD+ 32D cells. Although the number of FLT3/ITD+ 32D cells declined after incubation with the FLT3/ITD inhibitor AC220, the cells became refractory to AC220, concomitant with up-regulation of RUNX1. Silencing of RUNX1 abrogated the emergence and proliferation of AC220-resistant FLT3/ITD+ 32D cells in the presence of AC220. Our data indicate that FLT3/ITD deregulates cell proliferation and differentiation and confers resistance to AC220 by up-regulating RUNX1 expression. These findings suggest an oncogenic role for RUNX1 in FLT3/ITD+ cells and that inhibition of RUNX1 function represents a potential therapeutic strategy in patients with refractory FLT3/ITD+ AML.

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Acknowledgments

This work was supported by research support funds from the Grant-in-Aid for Scientific Research (20390298 and 25461593 to S.F.) and a Grant-in-Aid for Young Investigators (15K19616 to T.H.) from the Japanese Society for the Promotion of Science.

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Authors and Affiliations

  1. Department of Pediatrics, Shimane University School of Medicine, 89-1 Enya-cho, Izumo, Shimane, 693-8501, Japan

    Tomohiro Hirade, Mariko Abe, Takeshi Taketani, Seiji Yamaguchi & Seiji Fukuda

  2. Department of Oncology/Hematology, Shimane University School of Medicine, Izumo, Japan

    Chie Onishi

  3. Division of Blood Transfusion, Shimane University School of Medicine, Izumo, Japan

    Takeshi Taketani

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  1. Tomohiro Hirade
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  2. Mariko Abe
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Correspondence to Tomohiro Hirade or Seiji Fukuda.

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Hirade, T., Abe, M., Onishi, C. et al. Internal tandem duplication of FLT3 deregulates proliferation and differentiation and confers resistance to the FLT3 inhibitor AC220 by Up-regulating RUNX1 expression in hematopoietic cells. Int J Hematol 103, 95–106 (2016). https://doi.org/10.1007/s12185-015-1908-8

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  • DOI: https://doi.org/10.1007/s12185-015-1908-8

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