Abstract

Cellular senescence is a stress-responsive cell-cycle arrest program that terminates the further expansion of (pre-)malignant cells1,2. Key signalling components of the senescence machinery, such as p16INK4a, p21CIP1 and p53, as well as trimethylation of lysine 9 at histone H3 (H3K9me3), also operate as critical regulators of stem-cell functions (which are collectively termed ‘stemness’)3. In cancer cells, a gain of stemness may have profound implications for tumour aggressiveness and clinical outcome. Here we investigated whether chemotherapy-induced senescence could change stem-cell-related properties of malignant cells. Gene expression and functional analyses comparing senescent and non-senescent B-cell lymphomas from Eμ-Myc transgenic mice revealed substantial upregulation of an adult tissue stem-cell signature, activated Wnt signalling, and distinct stem-cell markers in senescence. Using genetically switchable models of senescence targeting H3K9me3 or p53 to mimic spontaneous escape from the arrested condition, we found that cells released from senescence re-entered the cell cycle with strongly enhanced and Wnt-dependent clonogenic growth potential compared to virtually identical populations that had been equally exposed to chemotherapy but had never been senescent. In vivo, these previously senescent cells presented with a much higher tumour initiation potential. Notably, the temporary enforcement of senescence in p53-regulatable models of acute lymphoblastic leukaemia and acute myeloid leukaemia was found to reprogram non-stem bulk leukaemia cells into self-renewing, leukaemia-initiating stem cells. Our data, which are further supported by consistent results in human cancer cell lines and primary samples of human haematological malignancies, reveal that senescence-associated stemness is an unexpected, cell-autonomous feature that exerts its detrimental, highly aggressive growth potential upon escape from cell-cycle blockade, and is enriched in relapse tumours. These findings have profound implications for cancer therapy, and provide new mechanistic insights into the plasticity of cancer cells.

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Acknowledgements

We thank G. Evan, the late A. Harris, T. Jacks and T. Jenuwein for mice, cells and materials; E. Berg, N. Burbach, A. Herrmann, H. Lammert, S. Mende, B. Teichmann and the Berlin-Brandenburg Center for Regenerative Therapies (BCRT) flow cytometry laboratory for technical assistance; and members of the Schmitt laboratory for discussions and editorial advice. This work was supported by a Ph.D. fellowship to J.R.D. from the Boehringer Ingelheim Foundation; by grants from the Deutsche Forschungsgemeinschaft to B.D., M.H. and C.A.S. (SFB/TRR 54) and to A.T. (SFB 873); by the Helmholtz Alliance ‘Preclinical Comprehensive Cancer Center’ grant (HA-305) from the Helmholtz Association to A.T. and C.A.S.; by the Dietmar Hopp Foundation to A.T.; and by the Deutsche Krebshilfe (grant 110678) to C.A.S. This interdisciplinary work was further made possible by the Berlin School of Integrative Oncology (BSIO) graduate program funded within the German Excellence Initiative, and the German Cancer Consortium (GCC).

Author information

Affiliations

  1. Charité – Universitätsmedizin Berlin, Medical Department of Hematology, Oncology and Tumor Immunology, and Molekulares Krebsforschungszentrum – MKFZ, Virchow Campus, 13353 Berlin, Germany

    • Maja Milanovic
    • , Dorothy N. Y. Fan
    • , Dimitri Belenki
    • , J. Henry M. Däbritz
    • , Jan R. Dörr
    • , Marlen Metzner
    • , Katharina Pardon
    • , Maurice Reimann
    • , Bernd Dörken
    • , Soyoung Lee
    •  & Clemens A. Schmitt
  2. Deutsches Konsortium für Translationale Krebsforschung (German Cancer Consortium), 69120 Heidelberg, Germany

    • Dorothy N. Y. Fan
    • , Andreas Trumpp
    • , Bernd Dörken
    • , Michael Hummel
    • , Soyoung Lee
    •  & Clemens A. Schmitt
  3. German Cancer Research Center (Deutsches Krebsforschungszentrum – DKFZ), Im Neuenheimer Feld 280, 69120 Heidelberg, Germany

    • Dorothy N. Y. Fan
    •  & Andreas Trumpp
  4. Deutsches Konsortium für Translationale Krebsforschung (German Cancer Consortium), Partner site Berlin, Berlin, Germany

    • Dorothy N. Y. Fan
    • , Andreas Trumpp
    • , Bernd Dörken
    • , Michael Hummel
    • , Soyoung Lee
    •  & Clemens A. Schmitt
  5. Cancer Biology and Genetics Program, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, New York 10065, USA

    • Zhen Zhao
  6. Max-Delbrück-Center for Molecular Medicine in the Helmholtz Association, Robert-Rössle-Straße 10, 13125 Berlin, Germany

    • Yong Yu
    • , Tamara Kanashova
    • , Bernd Dörken
    • , Gunnar Dittmar
    • , Soyoung Lee
    •  & Clemens A. Schmitt
  7. Charité – Universitätsmedizin Berlin, Department of Pathology, Berlin, Germany

    • Lora Dimitrova
    • , Dido Lenze
    •  & Michael Hummel
  8. Institute of Molecular Pathology (IMP), Vienna Biocenter, Dr Bohr-Gasse 7, 1030 Vienna, Austria

    • Ines A. Monteiro Barbosa
    •  & Johannes Zuber
  9. Equipe Labellisée Ligue Contre le Cancer, Department of Functional Genomics and Cancer, Institut de Génétique et de Biologie Moléculaire et Cellulaire, Centre National de la Recherche Scientifique, UMR7104, Institut National de la Santé et de la Recherche Médicale, U964, Université de Strasbourg, 67400 Illkirch, France

    • Marco A. Mendoza-Parra
    •  & Hinrich Gronemeyer
  10. Heidelberg Institute for Stem Cell Technology and Experimental Medicine (HI-STEM gGmbH), Im Neuenheimer Feld 280, 69120 Heidelberg, Germany

    • Andreas Trumpp
  11. Berlin Institute of Health, Anna-Louisa-Karsch-Straße 2, 10178 Berlin, Germany

    • Bernd Dörken
    • , Michael Hummel
    •  & Clemens A. Schmitt
  12. Luxembourg Institute of Health, 1A-B rue Thomas Edison, L-1455 Strassen, Luxembourg

    • Gunnar Dittmar

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Contributions

M.Mi. performed mouse lymphoma and leukaemia work, stem-cell and senescence assays, and gene set enrichment analyses. J.H.M.D. and M.R. conducted analyses with human cancer cell lines and primary human material. D.N.Y.F. and D.B. carried out flow cytometric analyses. Z.Z. generated leukaemias in the p53-regulatable mouse T-ALL model, I.A.M.B. and J.Z. in the p53-regulatable mouse AML model. Y.Y. carried out biochemical analyses. J.R.D. provided transcriptome analyses and long-term outcome data from senescence-capable mouse lymphomas. L.D. and M.A.M.-P. performed chromatin immunoprecipitations and analysed related datasets. D.L. conducted Affymetrix gene expression profiling and analyses. T.K. and G.D. carried out proteome analyses. M.Me. generated β-catenin/TCF-reporter cancer cell lines and performed luciferase reporter assays. K.P. generated qPCR data. A.T., B.D., H.G. and S.L. contributed to study design, data interpretation and preparation of the manuscript. M.H. provided immunohistochemical analyses. C.A.S. designed experiments, analysed the data and wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Clemens A. Schmitt.

Reviewer Information Nature thanks J. P. Medema, J. Vormoor and the other anonymous reviewer(s) for their contribution to the peer review of this work.

Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

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    This file contains supplementary data figure 1; showing the complete western blot membrane scans corresponding to extended data figures 2, 5, 6 and 8.

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