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Excess of NPM-ALK oncogenic signaling promotes cellular apoptosis and drug dependency

Oncogene volume 35, pages 3854–3865 (2016)Cite this article

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Abstract

Most of the anaplastic large-cell lymphoma (ALCL) cases carry the t(2;5; p23;q35) that produces the fusion protein NPM-ALK (nucleophosmin-anaplastic lymphoma kinase). NPM-ALK-deregulated kinase activity drives several pathways that support malignant transformation of lymphoma cells. We found that in ALK-rearranged ALCL cell lines, NPM-ALK was distributed in equal amounts between the cytoplasm and the nucleus. Only the cytoplasmic portion was catalytically active in both cell lines and primary ALCL, whereas the nuclear portion was inactive because of heterodimerization with NPM1. Thus, about 50% of the NPM-ALK is not active and sequestered as NPM-ALK/NPM1 heterodimers in the nucleus. Overexpression or relocalization of NPM-ALK to the cytoplasm by NPM genetic knockout or knockdown caused ERK1/2 (extracellular signal-regulated protein kinases 1 and 2) increased phosphorylation and cell death through the engagement of an ATM/Chk2- and γH2AX (phosphorylated H2A histone family member X)-mediated DNA-damage response. Remarkably, human NPM-ALK-amplified cell lines resistant to ALK tyrosine kinase inhibitors (TKIs) underwent apoptosis upon drug withdrawal as a consequence of ERK1/2 hyperactivation. Altogether, these findings indicate that an excess of NPM-ALK activation and signaling induces apoptosis via oncogenic stress responses. A ‘drug holiday’ where the ALK TKI treatment is suspended could represent a therapeutic option in cells that become resistant by NPM-ALK amplification.

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Acknowledgements

We thank Maria Stella Scalzo for technical support, Dr Emanuela Colombo for kindly providing MEFs that lack NPM1 (MEF NPM−/−p53−/−) and control fibroblasts (MEF p53−/−), Dr Guido Serini for the use of his confocal microscopy unit at the Candiolo Cancer Institute—IRCCS, Torino, Italy. We also thank Ariad Pharmaceutical, Pfizer, Astellas and Novartis that kindly provided all drugs used in this study. This work was supported by the Regione Lombardia (ID14546A) and Fondazione Berlucchi Onlus Grant 2014 (to CGP), and by grants FP7 ERC-2009-StG (Proposal No. 242965—‘Lunely’); Associazione Italiana per la Ricerca sul Cancro (AIRC) Grant IG-12023; Koch Institute/DFCC Bridge Project Fund; Ellison Foundation Boston; Worldwide Cancer Research Association (former AICR) grant 12-0216; the Grant for Oncology Innovation by Merck-Serono and R01 CA196703-01 (to RC).

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Author notes

  1. M Ceccon and M E Boggio Merlo: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Health Science, University of Milano-Bicocca, Monza, Italy

    M Ceccon, L Mologni, S Bombelli & C Gambacorti-Passerini

  2. Department of Molecular Biotechnology and Health Sciences, University of Torino, Torino, Italy

    M E Boggio Merlo, T Poggio, L M Varesio, M Menotti, A D Manazza, F Di Giacomo, C Mastini, M Compagno, R Chiarle & C Voena

  3. Center for Experimental Research and Medical Studies (CERMS), Città della Salute e della Scienza, Torino, Italy

    M E Boggio Merlo, T Poggio, L M Varesio, M Menotti, A D Manazza, F Di Giacomo, C Mastini, M Compagno, R Chiarle & C Voena

  4. Surgery and Translational Medicine Department, University of Milano-Bicocca, Monza, Italy

    R Rigolio

  5. Molecular Oncology Program, Centro Nacional de Investigaciones Oncológicas, Madrid, Spain

    C Ambrogio

  6. Tettamanti Research Centre, Pediatric Clinic, University of Milano-Bicocca, Monza, Italy

    G Giudici

  7. MetaSystems s.l.r., Milano, Italy

    C Casati

  8. Department of Pathology, Children’s Hospital and Harvard Medical School, Boston, MA, USA

    M Compagno & R Chiarle

  9. Division of Molecular Histopathology, Addenbrooke's Hospital Cambridge, Cambridge, UK

    S D Turner

  10. Section of Haematology, San Gerardo Hospital, Monza, Italy

    C Gambacorti-Passerini

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  1. M Ceccon
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Correspondence to C Gambacorti-Passerini or R Chiarle.

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Ceccon, M., Merlo, M., Mologni, L. et al. Excess of NPM-ALK oncogenic signaling promotes cellular apoptosis and drug dependency. Oncogene 35, 3854–3865 (2016). https://doi.org/10.1038/onc.2015.456

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