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Acute Leukemias

Activated Notch counteracts Ikaros tumor suppression in mouse and human T-cell acute lymphoblastic leukemia

Leukemia volume 29, pages 1301–1311 (2015)Cite this article

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Abstract

Activating NOTCH1 mutations occur in ~60% of human T-cell acute lymphoblastic leukemias (T-ALLs), and mutations disrupting the transcription factor IKZF1 (IKAROS) occur in ~5% of cases. To investigate the regulatory interplay between these driver genes, we have used a novel transgenic RNA interference mouse model to produce primary T-ALLs driven by reversible Ikaros knockdown. Restoring endogenous Ikaros expression in established T-ALL in vivo acutely represses Notch1 and its oncogenic target genes including Myc, and in multiple primary leukemias causes disease regression. In contrast, leukemias expressing high levels of endogenous or engineered forms of activated intracellular Notch1 (ICN1) resembling those found in human T-ALL rapidly relapse following Ikaros restoration, indicating that ICN1 functionally antagonizes Ikaros in established disease. Furthermore, we find that IKAROS mRNA expression is significantly reduced in a cohort of primary human T-ALL patient samples with activating NOTCH1/FBXW7 mutations, but is upregulated upon acute inhibition of aberrant NOTCH signaling across a panel of human T-ALL cell lines. These results demonstrate for the first time that aberrant NOTCH activity compromises IKAROS function in mouse and human T-ALL, and provide a potential explanation for the relative infrequency of IKAROS gene mutations in human T-ALL.

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Acknowledgements

We thank Mathew Salzone, Melanie Salzone, M Dayton, E Lanera, G Dabrowski, P Kennedy, K Stoev, C Smith, L Wilkins, S Brown and WEHI Bioservices staff for mouse work; W Alexander and E Major for ES cell and mouse resources; R Lane, J Corbin and A Keniry for technical assistance; E Viney and J Sarkis at the Australian Phenomics Network Transgenic RNAi service; M Everest and M Tinning at the Australian Genome Research Facility; and W Shi for assistance with exactSNP. We also thank the Children’s Oncology Group for primary human T-ALL samples, S Lowe and J Zuber for vectors and D Largaespada for Vav-tTA mice, and also S Lowe, J Zuber, D Izon, N Kershaw and members of the Dickins laboratory for advice and discussions. This work was supported by the National Health and Medical Research Council of Australia Project Grants 575535 and 1024599, Program Grant 490037, Senior Research Fellowship (GKS), Career Development Fellowship (RAD) and Early Career Fellowship (LC). IA was supported by the National Institutes of Health (1RO1CA133379, 1RO1CA105129, 1RO1CA149655, 5RO1CA173636 and 5RO1CA169784), the William Lawrence and Blanche Hughes Foundation, The Leukemia & Lymphoma Society, The V Foundation for Cancer Research and the St Baldrick’s Foundation. The work was also funded by Australian Government NHMRC IRIISS, an Australian Research Council Future Fellowship (SLN), Boehringer Ingelheim (MB), an ERC Advanced Grant (291740-LymphoControl) from the European Community’s Seventh Framework Program (MB), the Leukaemia Foundation of Australia (scholarship to MW, fellowship to MDM), a Sylvia and Charles Viertel Charitable Foundation Fellowship (RAD), Victorian State Government OIS grants and a Victorian Endowment for Science, Knowledge and Innovation (VESKI) Fellowship (RAD).

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Author notes

  1. M T Witkowski and L Cimmino: These authors contributed equally to this work.

Authors and Affiliations

  1. Molecular Medicine Division, Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia

    M T Witkowski, L Cimmino, M D McKenzie, S A Best, L Tuohey, T A Willson & R A Dickins

  2. Department of Medical Biology, University of Melbourne, Parkville, VIC, Australia

    M T Witkowski, L Cimmino, M D McKenzie, S A Best, L Tuohey, T A Willson, S L Nutt & R A Dickins

  3. Department of Pathology, NYU School of Medicine, New York, NY, USA

    L Cimmino, T Trimarchi & I Aifantis

  4. Bioinformatics Division, Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia

    Y Hu & G K Smyth

  5. Research Institute of Molecular Pathology, Vienna Biocenter, Vienna, Austria

    H Tagoh & M Busslinger

  6. Molecular Immunology Division, Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia

    S L Nutt

  7. Department of Mathematics and Statistics, University of Melbourne, Parkville, VIC, Australia

    G K Smyth

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  1. M T Witkowski
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Witkowski, M., Cimmino, L., Hu, Y. et al. Activated Notch counteracts Ikaros tumor suppression in mouse and human T-cell acute lymphoblastic leukemia. Leukemia 29, 1301–1311 (2015). https://doi.org/10.1038/leu.2015.27

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