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Chronic Myeloproliferative Neoplasias

Gab2 signaling in chronic myeloid leukemia cells confers resistance to multiple Bcr-Abl inhibitors

Leukemia volume 27, pages 118–129 (2013)Cite this article

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Abstract

Grb2-associated binder 2 (Gab2) serves as a critical amplifier in the signaling network of Bcr-Abl, the driver of chronic myeloid leukemia (CML). Despite the success of tyrosine kinase inhibitors (TKIs) in CML treatment, TKI resistance, caused by mutations in Bcr-Abl or aberrant activity of its network partners, remains a clinical problem. Using inducible expression and knockdown systems, we analyzed the role of Gab2 in Bcr-Abl signaling in human CML cells, especially with respect to TKI sensitivity. We show for the first time that Gab2 signaling protects CML cells from various Bcr-Abl inhibitors (imatinib, nilotinib, dasatinib and GNF-2), whereas Gab2 knockdown or haploinsufficiency leads to increased TKI sensitivity. We dissected the underlying molecular mechanism using various Gab2 mutants and kinase inhibitors and identified the Shp2/Ras/ERK and the PI3K/AKT/mTOR axes as the two critical signaling pathways. Gab2-mediated TKI resistance was associated with persistent phosphorylation of Gab2 Y452, a PI3K recruitment site, and consistent with this finding, the protective effect of Gab2 was completely abolished by the combination of dasatinib with the dual PI3K/mTOR inhibitor NVP-BEZ235. The identification of Gab2 as a novel modulator of TKI sensitivity in CML suggests that Gab2 could be exploited as a biomarker and therapeutic target in TKI-resistant disease.

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Acknowledgements

We thank Dr Alexander Kühnemund and members of the Brummer laboratory for helpful discussions, Andreas Geissler for advice on the caspase assays and Katja Thurig, Christine Capri and Jennifer Schwarz for technical assistance. TB is supported by the German Research Foundation (DFG) through the Emmy-Noether-Program (BR3662/1-1) and EXC 294 BIOSS. FUW is funded by the Excellence Initiative of the DFG (GSC-4; Spemann Graduate School for Biology and Medicine) and EXC 294 BIOSS. RZ is supported by the DFG (ZE 872/1–1 and Heisenberg Fellowship ZE 872/2–1). SL, MW, RZ and TB are also supported by the DFG funded Collaborative Research Center 850. HLP gratefully acknowledges support by the DFG (Pa 611/5-1 and Pa 611/6-1) as well as the NIH (PO1 CA108671). RJD is supported by the National Health and Medical Research Council of Australia. DS is supported by the Austrian Academy of Sciences through an APART fellowship. KA is supported by a fellowship of the medical faculty of the University of Freiburg.

Author contributions

FUW, SH, SS and SB performed all cellular and biochemical experiments. KA conducted the IHC analyses. KA, SL, HLP, MW, CFW and RZ provided clinical samples and expertise. PA, DS and JMP provided critical reagents and mice. All authors contributed to the design, analysis and discussion of experiments. TB wrote the manuscript together with FUW, SH and RJD. All authors reviewed the manuscript.

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Authors and Affiliations

  1. Centre for Biological Systems Analysis (ZBSA), Albert-Ludwigs-University of Freiburg, Freiburg, Germany

    F U Wöhrle, S Halbach, S Braun & T Brummer

  2. Faculty of Biology, Institute for Biology III, Albert-Ludwigs-University of Freiburg, Freiburg, Germany

    F U Wöhrle, S Halbach, S Braun & T Brummer

  3. Spemann Graduate School of Biology and Medicine, Albert-Ludwigs-University of Freiburg, Freiburg, Germany

    F U Wöhrle & S Halbach

  4. Centre for Biological Signaling Studies BIOSS, Albert-Ludwigs-University of Freiburg, Freiburg, Germany

    F U Wöhrle, S Laßmann, R Zeiser & T Brummer

  5. Department of Pathology, Oncology University Hospital Freiburg, Freiburg, Germany

    K Aumann, S Laßmann & M Werner

  6. Department of Hematology, Oncology University Hospital Freiburg, Freiburg, Germany

    S Schwemmers, C F Waller, H L Pahl & R Zeiser

  7. INSERM U895, Centre Méditerranéen de Médecine Moléculaire, Team Cell Death, Differentiation, Inflammation and Cancer, Nice, France

    P Auberger

  8. Laboratory of Mammalian Cell Biology and Development, The Rockefeller University, New York, NY, USA

    D Schramek

  9. Institute of Molecular Biotechnology of the Austrian Academy of Sciences, Vienna, Austria

    J M Penninger

  10. Comprehensive Cancer Center Freiburg (CCCF), University Hospital Freiburg, Freiburg, Germany

    S Laßmann, M Werner, C F Waller, H L Pahl, R Zeiser & T Brummer

  11. Garvan Institute of Medical Research, Sydney, New South Wales, Australia

    R J Daly

  12. Department of Medicine, St Vincent’s Hospital Clinical School, University of New South Wales, Sydney, New South Wales, Australia

    R J Daly

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  1. F U Wöhrle
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  2. S Halbach
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Wöhrle, F., Halbach, S., Aumann, K. et al. Gab2 signaling in chronic myeloid leukemia cells confers resistance to multiple Bcr-Abl inhibitors. Leukemia 27, 118–129 (2013). https://doi.org/10.1038/leu.2012.222

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