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Myelodysplastic syndrome

Molecular characterization of EZH2 mutant patients with myelodysplastic/myeloproliferative neoplasms

Leukemia volume 31, pages 1936–1943 (2017)Cite this article

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Abstract

Mutations in the epigenetic regulator gene EZH2 are frequently observed in patients with myelodysplastic/myeloproliferative neoplasms (MDS/MPN; 10–13%) and are associated with a poor outcome. To gain more insight into EZH2 pathology, we sought to genetically characterize a cohort of 41 EZH2-mutated MDS/MPN patients using targeted deep next-generation sequencing (NGS), colony-forming progenitor assays and transcriptome analysis. Stable short hairpin RNA (shRNA)-mediated downregulation of EZH2 was performed in MDS-derived F-36P, MOLM-13 and OCI-M2 cells to study EZH2-specific changes. Targeted NGS revealed a complex pattern of mutations with a total of 190 individual mutations. EZH2 mutations frequently co-occur with TET2 (58%), RUNX1 (40%) and ASXL1 (34%) mutations. Colony assays indicated EZH2 mutations to be mostly early events in leukemogenesis and showed a complex mutational hierarchy. Gene expression data revealed a number of differently expressed genes between EZH2 wild-type and mutant patients including known EZH2 targets. Comparison of patient transcriptome to EZH2-downregulated cell line data revealed several genes as novel EZH2 targets, showing opposite as well as unidirectional regulation between cell lines and patients. Some genes, such as CXXC5, ETS1 and VAV3 have previously been implied to have a role in leukemogenesis. Their precise role in MDS/MPN needs to be further investigated.

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Acknowledgements

The excellent technical assistance of Mrs Anja Waldau is gratefully acknowledged. The study was supported by the Deutsche José Carreras Leukämie-Stiftung e.V. (DJCLS R15/20) and the Interdisziplinäres Zentrum für Klinische Forschung (IZKF, Jena, Germany).

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Authors and Affiliations

  1. Abteilung Hämatologie und Internistische Onkologie, Klinik für Innere Medizin II, Universitätsklinikum Jena, Jena, Germany

    J Rinke, V Schäfer, N Winkelmann, A Hochhaus & T Ernst

  2. Institut für Molekulare Zellbiologie, Center for Molecular Biomedicine, Universitätsklinikum Jena, Jena, Germany

    J P Müller

  3. Klinik für Anästhesiologie und Intensivmedizin, Universitätsklinikum Jena, Jena, Germany

    M F Blaess

  4. National Genetics Reference Laboratory (Wessex), Salisbury District Hospital, Salisbury, UK

    A Chase & N C P Cross

  5. Faculty of Medicine, University of Southampton, Southampton, UK

    A Chase & N C P Cross

  6. MLL Münchner Leukämielabor, Munich, Germany

    M Meggendorfer & C Haferlach

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  1. J Rinke
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Correspondence to T Ernst.

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Rinke, J., Müller, J., Blaess, M. et al. Molecular characterization of EZH2 mutant patients with myelodysplastic/myeloproliferative neoplasms. Leukemia 31, 1936–1943 (2017). https://doi.org/10.1038/leu.2017.190

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