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Mechanisms of resistance

F604S exchange in FIP1L1-PDGFRA enhances FIP1L1-PDGFRA protein stability via SHP-2 and SRC: a novel mode of kinase inhibitor resistance

Leukemia volume 29, pages 1763–1770 (2015)Cite this article

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Abstract

FIP1L1-PDGFRA is a constitutively activated kinase described in chronic eosinophilic leukemia (CEL) and hypereosinophilic syndrome (HES). Imatinib is clinically active in FIP1L1-PDGFRA-positive diseases. Using in vitro screening to identify imatinib-resistant mutations, we frequently detected a Phe to Ser exchange at position 604 (F604S) of FIP1L1-PDGFRA alone or in combination with other exchanges. Surprisingly, FIP1L1-PDGFRA/F604S did not increase the biochemical or cellular IC50 value of imatinib when compared with unmutated FIP1L1-PDGFRA. However, FIP1L1-PDGFRA/F604S more efficiently induced growth factor independence in cell lines and primary mouse bone marrow cells. Pulse chase analysis revealed that the F604S exchange strongly stabilized FIP1L1-PDGFRA/F604S. The F604S mutation creates a binding site for the phosphatase domain of SHP-2, leading to lower autophosphorylation of FIP1L1-PDGFRA/F604S. This is associated with a reduced activation of SRC and CBL by FIP1L1-PDGFRA/F604S compared with the unmutated oncogene. As SRC inhibition and knockdown resulted in FIP1L1-PDGFRA stabilization, this explains the extended half-life of FIP1L1-PDGFRA/F604S. Interestingly, FIP1L1-PDGFRA/L629P, a recently identified mutation in an imatinib-resistant CEL patient, also showed protein stabilization similar to that observed with FIP1L1-PDGFRA/F604S. Therefore, resistance mutations in FIP1L1-PDGFRA that do not interfere with drug binding but rather increase target protein stability seem to be one of the drug-resistance mechanisms in FIP1L1-PDGFRA-positive disease.

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Acknowledgements

JD is supported by a grant from the Jose Carreras Leukämie-Stiftung (DJCLS R 14/22). We thank Dr N. Cross, Wessex Regional Genetics Laboratory, Salisbury, UK, for providing information about drug-resistant FP-positive patients.

Author Contributions

SPG: Conceived the study, designed and performed the experiments, analyzed data and wrote the manuscript. KZ, CY, ALI, NB, AR: analyzed the data. JD: Conceived the study, analyzed data and wrote the manuscript. All authors read and approved the final manuscript.

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Author notes

  1. N Von Bubnoff and J Duyster: These authors share senior authorship.

Authors and Affiliations

  1. Department of Internal Medicine I, University Medical Center Freiburg, Freiburg, Germany

    S P Gorantla, K Zirlik, C Yu, A L Illert, N Von Bubnoff & J Duyster

  2. German Cancer Consortium (DKTK), Freiburg and German Cancer Research Center (DKFZ), Heidelberg, Germany

    S P Gorantla & J Duyster

  3. Department of Hematology and Oncology, University Hospital Mannheim, Mannheim, Germany

    A Reiter

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  1. S P Gorantla
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Correspondence to J Duyster.

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Gorantla, S., Zirlik, K., Reiter, A. et al. F604S exchange in FIP1L1-PDGFRA enhances FIP1L1-PDGFRA protein stability via SHP-2 and SRC: a novel mode of kinase inhibitor resistance. Leukemia 29, 1763–1770 (2015). https://doi.org/10.1038/leu.2015.70

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