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Acute Leukemias

Pim kinases phosphorylate Chk1 and regulate its functions in acute myeloid leukemia

Leukemia volume 28, pages 293–301 (2014)Cite this article

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Abstract

Phosphorylation by Akt on Ser 280 was reported to induce cytoplasmic retention and inactivation of CHK1 with consequent genetic instability in PTEN−/− cells. In acute myeloid leukemia cells carrying the FLT3-internal tandem duplication (ITD) mutation, we observed high rates of FLT3-ITD-dependent CHK1 Ser 280 phosphorylation. Pharmacological inhibition and RNA interference identified Pim1/2, not Akt, as effectors of this phosphorylation. Pim1 catalyzed Ser 280 phosphorylation in vitro and ectopic expression of Pim1/2-induced CHK1 phosphorylation. Ser 280 phosphorylation did not modify CHK1 localization, but facilitated its cell cycle and resistance functions in leukemic cells. FLT3, PIM or CHK1 inhibitors synergized with DNA-damaging agents to induce apoptosis, allowing cells to bypass the etoposide-induced G2/M arrest. Consistently, etoposide-induced CHK1-dependent phosphorylations of CDC25C on Ser 216 and histone H3 on Thr11 were decreased upon FLT3 inhibition. Accordingly, ectopic expression of CHK1 improved the resistance of FLT3-ITD cells and maintained histone H3 phosphorylation in response to DNA damage, whereas expression of unphosphorylated Ser 280Ala mutant did not. Finally, FLT3- and Pim-dependent phosphorylation of CHK1 on Ser 280 was confirmed in primary blasts from patients. These results identify a new pathway involved in the resistance of FLT3-ITD leukemic cells to genotoxic agents, and they constitute the first report of CHK1 Ser 280 regulation in myeloid malignancies.

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Acknowledgements

We are very grateful to Dr Cécile Demur and Professor Eric Delabesse for managing and characterization of primary samples. We also thank Dr Shuchi Agrawal-Singh (Copenhagen University) and Dr Ramon Parsons (Columbia University), for kindly providing us with the Pim and CHK1 expression plasmids respectively. We thank Fatima L’Faqihi for technical assistance at the cytometry and cell sorting facility of INSERM 1043, Toulouse. This work was supported by the Association GAEL (Gael Adolescent Espoir Leucémie), by the Institut National de la Santé et de la Recherche Médicale (INSERM), by the Centre National de la Recherche Scientifique (CNRS), by the Association pour la Recherche contre le Cancer (ARC, grant SFI20101201865), by the Institut National du Cancer (INCA, PL2008 INCA-Gov-1345), by the association Inna Biosanté, by the Laboratoire d'Excellence Toulouse Cancer LABEX TOUCAN (Integrative analysis of resistance in hematological cancers) and by the association Laurette Fugain.

Author Contributions

Ling Li Yuan is a recipient of the China Scholarship Council and of the Société Française d’Hématologie (SFH).

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Authors and Affiliations

  1. Cancer Research Center of Toulouse, Inserm Unité Mixte de Recherche 1037, CNRS Equipe de Recherche labellisée 5294, Université de Toulouse, Centre Hospitalier Universitaire Purpan, Toulouse, France

    L L Yuan, S Bertoli, F Grimal, V Mansat-De Mas, C Dozier, C Récher, C Didier & S Manenti

  2. Institut Cochin, Université Paris Descartes, CNRS UMR 8104, INSERM U 1016, Paris, France

    A S Green & J Tamburini

  3. Service d’Hématologie, Centre Hospitalier Universitaire Purpan, Toulouse, France

    S Bertoli & C Récher

  4. Laboratoire d’Hématologie, Centre Hospitalier Universitaire Purpan, Toulouse, France

    V Mansat-De Mas

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  1. L L Yuan
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Correspondence to S Manenti.

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Yuan, L., Green, A., Bertoli, S. et al. Pim kinases phosphorylate Chk1 and regulate its functions in acute myeloid leukemia. Leukemia 28, 293–301 (2014). https://doi.org/10.1038/leu.2013.168

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