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Mutant nucleophosmin and cooperating pathways drive leukemia initiation and progression in mice

Nature Genetics volume 43, pages 470–475 (2011)Cite this article

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Abstract

Acute myeloid leukemia (AML) is a molecularly diverse malignancy with a poor prognosis whose largest subgroup is characterized by somatic mutations in NPM1, which encodes nucleophosmin1. These mutations, termed NPM1c, result in cytoplasmic dislocation of nucleophosmin1 and are associated with distinctive transcriptional signatures2, yet their role in leukemogenesis remains obscure. Here we report that activation of a humanized Npm1c knock-in allele in mouse hemopoietic stem cells causes Hox gene overexpression, enhanced self renewal and expanded myelopoiesis. One third of mice developed delayed-onset AML, suggesting a requirement for cooperating mutations. We identified such mutations using a Sleeping Beauty3,4 transposon, which caused rapid-onset AML in 80% of mice with Npm1c, associated with mutually exclusive integrations in Csf2, Flt3 or Rasgrp1 in 55 of 70 leukemias. We also identified recurrent integrations in known and newly discovered leukemia genes including Nf1, Bach2, Dleu2 and Nup98. Our results provide new pathogenetic insights and identify possible therapeutic targets in NPM1c+ AML.

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Figure 1: Conditional mouse model of type A NPM1c mutation.
Figure 2: Hematopoietic changes and incidence of AML in Npm1cA/+ mice.
Figure 3: Npm1cA and the GrOnc transposon synergize to cause AML.
Figure 4: Common integration sites in transposon-derived leukemias.
Figure 5: A model for Npm1cA/+-driven leukemogenesis.

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Acknowledgements

We acknowledge the use of the Research Support Facility at the Wellcome Trust Sanger Institute, the Department of Pathology Tissue Bank and the Cambridge National Institute of Health Research (NIHR) Biomedical Research Centre, University of Cambridge. We thank F. Law and J. Gadiot for assistance in generating the Npm1flox-CA and Rosaflox-SB mice; F. Foyer and B. Graham for help with fluorescent microscopy; B. Ling, W. Cheng, R. Macintyre and P. Chan for help with flow cytometry; R. Bautista for help with gene expression images; C. Hale and A. Nyzhnyk for help with ELISAs; B. Huntly, D. Adams, J. Cadinanos, H. Prosser, N. Conte, K. Yusa and Q. Liang for helpful discussions during the project; and P. Campbell and A. Green for critical reading of the manuscript. This work was supported by a Clinician Scientist Fellowship from Cancer Research UK (G.S.V.).

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Authors and Affiliations

  1. Mouse Genomics Team, The Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, UK.,

    George S Vassiliou, Jonathan L Cooper, Roland Rad, Stephen Rice, Lena Rad, Peter Ellis, Rob Andrews, Ruby Banerjee, Carolyn Grove, Wei Wang, Pentao Liu & Allan Bradley

  2. Department of Haematology, Cambridge Institute for Medical Research, University of Cambridge, Cambridge, UK

    Juan Li

  3. The Netherlands Cancer Institute, Amsterdam, The Netherlands

    Anthony Uren

  4. Department of Pathology, University of Cambridge, Addenbrooke's Hospital, Cambridge, UK

    Penny Wright & Mark Arends

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  1. George S Vassiliou
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Contributions

G.S.V. and A.B. designed the study. G.S.V. generated Npm1flox-cA mice, GrOnc mice and GFP-NPM1 constructs, managed mouse colonies, designed and validated polyclonal anti-Npm1c sera and carried out protein blots. J.L.C. and G.S.V. performed mouse genotyping, tumor processing and banking and K562 transfections. G.S.V., J.L.C., R.R. and L.R. performed mouse necropsies. G.S.V., J.L.C. and J.L. performed hemopoietic analyses. G.S.V. and C.G. performed quantitative PCR. G.S.V., P.E. and R.A. performed gene expression analysis studies. S.R., G.S.V. and R.R. performed mapping and analysis of transposon integration sites. G.S.V. and R.B. performed fluorescence in situ hybridization. W.W. and P.L. generated the Stella-Cre mice. A.U. generated the Rosaflox-SB mice. P.W. and M.A. performed histological analyses. A.B. supervised the study. All authors contributed to the writing of the manuscript.

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Correspondence to George S Vassiliou or Allan Bradley.

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Vassiliou, G., Cooper, J., Rad, R. et al. Mutant nucleophosmin and cooperating pathways drive leukemia initiation and progression in mice. Nat Genet 43, 470–475 (2011). https://doi.org/10.1038/ng.796

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