A novel tumour-suppressor function for the Notch pathway in myeloid leukaemia

Journal name:
Nature
Volume:
473,
Pages:
230–233
Date published:
DOI:
doi:10.1038/nature09999
Received
Accepted
Published online

Notch signalling is a central regulator of differentiation in a variety of organisms and tissue types1. Its activity is controlled by the multi-subunit γ-secretase (γSE) complex2. Although Notch signalling can play both oncogenic and tumour-suppressor roles in solid tumours, in the haematopoietic system it is exclusively oncogenic, notably in T-cell acute lymphoblastic leukaemia, a disease characterized by Notch1-activating mutations3. Here we identify novel somatic-inactivating Notch pathway mutations in a fraction of patients with chronic myelomonocytic leukaemia (CMML). Inactivation of Notch signalling in mouse haematopoietic stem cells (HSCs) results in an aberrant accumulation of granulocyte/monocyte progenitors (GMPs), extramedullary haematopoieisis and the induction of CMML-like disease. Transcriptome analysis revealed that Notch signalling regulates an extensive myelomonocytic-specific gene signature, through the direct suppression of gene transcription by the Notch target Hes1. Our studies identify a novel role for Notch signalling during early haematopoietic stem cell differentiation and suggest that the Notch pathway can play both tumour-promoting and -suppressive roles within the same tissue.

At a glance

Figures

  1. Ncstn deficiency leads to CMML-like disease and a significant enlargement of the GMP progenitor population.
    Figure 1: Ncstn deficiency leads to CMML-like disease and a significant enlargement of the GMP progenitor population.

    a, Histological analysis showing accumulation of monocytes and granulocytes in peripheral blood (Wright–Giemsa staining), spleen and liver (haematoxylin and eosin staining). A magnification of each infiltrant is shown in the lowest panels. b, Absolute numbers of each monocytic/granulocytic subset from the spleen of control and Ncstnf/f Vav-cre+ littermate animals (12 weeks of age, mean±s.d., n = 10). c, Absolute numbers of each progenitor subpopulation in the bone marrow (mean±s.d., n = 10). d, Detailed FACS analysis of bone marrow and spleen myeloid progenitor (myeloid progenitors: Lin/c-Kit+/Sca-1) populations of Ncstnf/f Vav-cre+ and Ncstnf/f Vav-cre littermates showing a significant enlargement of the GMP (FcγRII/III+, CD34+) subset.

  2. Notch signalling suppresses an extensive myeloid gene expression program through the induction of the transcriptional repressor Hes1.
    Figure 2: Notch signalling suppresses an extensive myeloid gene expression program through the induction of the transcriptional repressor Hes1.

    a, Heat map showing regulation of genes representative of the myeloid signature from the indicated cell populations and mice. b, Expression data were analysed for lists of genes positively involved in myelopoiesis using gene-set enrichment analysis. Enrichment plots show upregulation of myeloid-specific genes in Ncstnf/fMx1-cre+ and downregulation in Notch1IC Mx1-cre+ LSK cells (compared with WT counterparts). c, Purified cKit+ progenitors from WT and Ncstnf/fMx1-cre+ mice were transduced with retroviruses encoding Hes-1 or empty vector, subsequently plated on methylcellulose for 7days and analysed for expression of myeloid or megakaryocyte differentiation markers (Gr1, CD41). A representative of four experiments is shown.

  3. Ectopic expression of Notch1-IC is able to prevent CMML-like disease in Ncstn-/- mice.
    Figure 3: Ectopic expression of Notch1-IC is able to prevent CMML-like disease in Ncstn/ mice.

    a, PolyI:polyC-induced Notch1-IC expression in Ncstnf/flsl-N1-IC Mx1-Cre+ animals suppresses myeloid cells in spleen. b, Notch1-IC expression suppresses GMP progenitor population in bone marrow. c, Induction of cell death in WT GMP cells cultured in the presence (OP9-DL1-4) or absence (OP9) of Notch ligands. Cell death was measured by the combination of 7AAD and annexin V staining 48h after co-culture initiation. For ac, a representative of more than three experiments is shown.

  4. Novel, loss-of-function Notch pathway mutations in human CMML.
    Figure 4: Novel, loss-of-function Notch pathway mutations in human CMML.

    a, Sequence traces of identified Notch pathway mutations in tumours from patients with CMML but not in normal tissues show somatic origin. b, Comparison of the percentage of Notch pathway mutations in specimens from patients with CMML, myelofibrosis and polycythemia vera. The asterisk denotes that only verified somatic CMML mutations are included. c, OP9-DL1 co-culture of WT LSK cells infected with specified constructs. Analysis of the CD11b+ population was studied 14days after the initiation of the culture. d, A similar experiment as in c, using LSK Ncstn−/− progenitors infected with the specified constructs. In all cases, a representative of more than three experiments is shown.

Accession codes

Primary accessions

Gene Expression Omnibus

References

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Author information

  1. These authors contributed equally to this work.

    • Apostolos Klinakis &
    • Camille Lobry

Affiliations

  1. Biomedical Research Foundation, Academy of Athens, Athens, Greece

    • Apostolos Klinakis &
    • Argiris Efstratiadis
  2. Howard Hughes Medical Institute and Department of Pathology, New York University School of Medicine, New York, New York 10016, USA

    • Camille Lobry,
    • Philmo Oh,
    • Silvia Buonamici,
    • Severine Cathelin,
    • Thomas Trimarchi,
    • Elisa Araldi,
    • Cynthia Liu,
    • Sherif Ibrahim &
    • Iannis Aifantis
  3. Human Oncology and Pathogenesis Program and Leukemia Service, Department of Medicine, Memorial Sloan-Kettering Cancer Center, New York, New York 10016, USA

    • Omar Abdel-Wahab &
    • Ross L. Levine
  4. Department of Biostatistics and Computational Biology, Dana-Farber Cancer Institute, and Department of Biostatistics, Harvard School of Public Health, Boston, Massachusetts 02115, USA

    • Hiroshi Haeno &
    • Franziska Michor
  5. Department of Clinical Chemistry, Microbiology and Immunology, Ghent University Hospital, Ghent University, Ghent, Belgium

    • Inge van De Walle &
    • Tom Taghon
  6. Department of Leukemia, M.D. Anderson Cancer Center, Houston, Texas 77030, USA

    • Miroslav Beran
  7. Department of Pathology, NYU Cancer Institute and Center for Health Informatics and Bioinformatics, NYU Langone Medical Center, New York, New York 10016, USA

    • Jiri Zavadil
  8. Novartis Institutes for Biomedical Research, Cambridge, Massachusetts 02139, USA

    • Silvia Buonamici

Contributions

I.A., C.L. and A.K. conceived the study and designed all experiments. A.K. and A.E. helped with experimental planning and generated the Ncstnf/f mice. C.L. performed most of the mouse experiments and in vitro studies aided by P.O., S.B, S.C., T.T. and E.A. R.L.L, O.A.-W. and M.B. performed and analysed human leukaemia sample exon sequencing. H.H. and F.M. helped with disease modelling and computational analysis of disease progression. I.v.D.W. and T.T. performed the human stem cell differentiation assays. C.L. and S.I. analysed mouse disease pathology. J.Z. processed and analysed gene expression data.

Competing financial interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to:

The microarray data are deposited in Gene Expression Omnibus of the National Center for Biotechnical Information under accession numbers GSE27794, GSE27799 and GSE27811.

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Supplementary information

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  1. Supplementary Information (15.1M)

    This file contains Supplementary Figure 1-16 with legends and Supplementary Tables 1-5.

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