Abstract

Leukaemogenesis requires enhanced self-renewal, which is induced by oncogenes. The underlying molecular mechanisms remain incompletely understood. Here, we identified C/D box snoRNAs and rRNA 2′-O-methylation as critical determinants of leukaemic stem cell activity. Leukaemogenesis by AML1-ETO required expression of the groucho-related amino-terminal enhancer of split (AES). AES functioned by inducing snoRNA/RNP formation via interaction with the RNA helicase DDX21. Similarly, global loss of C/D box snoRNAs with concomitant loss of rRNA 2′-O-methylation resulted in decreased leukaemia self-renewal potential. Genomic deletion of either C/D box snoRNA SNORD14D or SNORD35A suppressed clonogenic potential of leukaemia cells in vitro and delayed leukaemogenesis in vivo. We further showed that AML1-ETO9a, MYC and MLL-AF9 all enhanced snoRNA formation. Expression levels of C/D box snoRNAs in AML patients correlated closely with in vivo frequency of leukaemic stem cells. Collectively, these findings indicate that induction of C/D box snoRNA/RNP function constitutes an important pathway in leukaemogenesis.

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Acknowledgements

We thank T. Gridley for providing Aes straight knockout (Aes−/−) mice. We are grateful to B. Edemir for help with project organization and manuscript preparation. We thank M. Scheller-Wendorff for support of the mice experiments. This work was supported by Deutsche Forschungsgemeinschaft (DFG, MU1328/15-1 and MU1328/9-2), the Deutsche Jose Carreras Leukämie-Stiftung e. V. (DJCLS R10/35f), by a Wilhelm-Roux-Program grant (28/43) of the Medical Faculty, Martin-Luther-University Halle-Wittenberg to CMT and the grants of the federal state of Saxony-Anhalt (P40). N.B. is supported by Wilhelm Sander Stiftung (2014.054.1), Deutsche Krebshilfe (70112282) and ‘Innovative Medical Research’ of the University of Münster Medical School (IMF) (121314 and 111501). C.P. is supported by a Max-Eder grant of the Deutsche Krebshilfe (70111531). The laboratory of R.S. is supported by Deutsche Forschungsgemeinschaft (DFG SL27/7-2). The laboratory of W.E.B. is supported by DFG grant EXC 1003. The work of 20 AML samples with LSC frequencies was supported by the Government of Canada through Genome Canada and the Ministère de l’enseignement supérieur, de la recherche, de la science et de la technologie du Québec through Génome Québec to G.S. and J.H. and by grants from the Cancer Research Network of the Fonds de recherche du Québec-Santé to J.H.

Author information

Affiliations

  1. Department of Hematology and Oncology, University of Halle, Halle 06120, Germany

    • Fengbiao Zhou
    • , Yi Liu
    • , Christian Rohde
    • , Cornelius Pauli
    • , Dennis Gerloff
    • , Chunhong Cui
    • , Stefanie Göllner
    • , Caroline Pabst
    •  & Carsten Müller-Tidow
  2. Department of Medicine V, Hematology, Oncology and Rheumatology, University of Heidelberg, Heidelberg 69120, Germany

    • Fengbiao Zhou
    • , Yi Liu
    • , Christian Rohde
    • , Stefanie Göllner
    • , Caroline Pabst
    •  & Carsten Müller-Tidow
  3. Institute of Molecular Medicine, University of Halle, Halle 06120, Germany

    • Marcel Köhn
    • , Danny Misiak
    •  & Stefan Hüttelmaier
  4. Department of Medicine A, Hematology and Oncology, University of Muenster, Muenster 48149, Germany

    • Nicole Bäumer
    •  & Wolfgang E. Berdel
  5. Department of Medicine II, Hematology/Oncology, Goethe University, Frankfurt Am Main 60590, Germany

    • Thomas Oellerich
    •  & Hubert Serve
  6. Department of Hematology, University of Cambridge, Hills Road, Cambridge CB2 0XY, UK

    • Thomas Oellerich
  7. German Cancer Research Center and German Cancer, Consortium, Heidelberg 69120, Germany

    • Thomas Oellerich
    •  & Hubert Serve
  8. Division of Genetics, Department of Biology, University of Erlangen-Nuremberg, Erlangen 91058, Germany

    • Maria-Paz Garcia-Cuellar
    •  & Robert Slany
  9. Department of Translational Hematology and Oncology Research, Taussig Cancer Institute, Cleveland, Ohio 44195, USA

    • Jaroslaw P. Maciejewski
    •  & Bartlomiej Przychodzen
  10. Institute of Medical Immunology, University of Halle, Halle 06120, Germany

    • Barbara Seliger
  11. Institute of Medical Informatics, University of Muenster, Muenster 48149, Germany

    • Hans-Ulrich Klein
    • , Christoph Bartenhagen
    •  & Martin Dugas
  12. Program in Medical and Population Genetics, Broad Institute, Cambridge, Massachusetts 02142, USA

    • Hans-Ulrich Klein
  13. Division of Experimental Therapeutics, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan

    • Makoto Mark Taketo
  14. Broad Institute, Cambridge, Massachusetts 02142, USA

    • Daneyal Farouq
    • , Schraga Schwartz
    •  & Aviv Regev
  15. Department of Biology, MIT, Cambridge, Massachusetts 02139, USA

    • Aviv Regev
  16. Howard Hughes Medical Institute, 4000 Jones Bridge Road, Chevy Chase, Maryland 20815, USA

    • Aviv Regev
  17. Division of Hematology-Oncology, Maisonneuve-Rosemont Hospital, Montreal, Quebec H1T 2M4, Canada

    • Josée Hébert
  18. Leukemia Cell Bank of Quebec, Maisonneuve-Rosemont Hospital, Montreal, Quebec H1T 2M4, Canada

    • Josée Hébert
    •  & Guy Sauvageau
  19. Department of Medicine, University of Montreal, Montreal, Quebec H3T 1J4, Canada

    • Josée Hébert
    •  & Guy Sauvageau
  20. Laboratory of Molecular Genetics of Stem Cells, Institute for Research in Immunology and Cancer, University of Montreal, Montreal, Quebec H3T 1J4, Canada

    • Guy Sauvageau

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Contributions

F.Z. designed and conducted the experiments, analysed the data and wrote the manuscript. Y.L. and C.P. made the snoRNA knockouts. M.K., D.M. and S.H. analysed snoRNA-seq data. T.O. and H.S. performed mass spectrometry analyses and analysed the data. C.P. helped with NSG mice experiments. M.-P.G.-C. and R.S. isolated nascent RNA. B.P., C.R., J.P.M., H.-U.K. C.B. and M.D. analysed NGS data. M.M.T. provided the Aesf/f mice. D.F., S.S. and A.R. performed pseudouridylation-sequencing. D.G., C.P., N.B., C.C., S.G., W.E.B. and B.S. discussed the results and advised on research. W.E.B. provided patient samples from the AML-AZA clinical study. J.H. and G.S. provided human AML specimens with determined LSC frequency. C.M.-T. designed and conceptualized the overall research, analysed the data, and wrote the manuscript. All authors read and approved the final manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Carsten Müller-Tidow.

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https://doi.org/10.1038/ncb3563

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