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ALKR1275Q perturbs extracellular matrix, enhances cell invasion and leads to the development of neuroblastoma in cooperation with MYCN

Oncogene volume 35, pages 4447–4458 (2016)Cite this article

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Abstract

Overexpression of MYCN is a hallmark of neuroblastoma (NB). ALKR1275Q, an activating mutation of ALK (anaplastic lymphoma kinase), has been found in sporadic and familial NB patients. In this report, we demonstrated that ALKR1275Q knock-in, MYCN transgenic compound mice developed NB with complete penetrance. Transcriptome analysis revealed that ALKR1275Q globally downregulated the expression of extracellular matrix (ECM)- and basement membrane (BM)-associated genes in both primary neuronal cells and NB tumors. Accordingly, ALKR1275Q/MYCN tumors exhibited reduced expression of ECM/BM-related proteins as compared with MYCN tumors. In addition, on MYCN transduction, ALKR1275Q-expressing neuronal cells exhibited increased migratory and invasive activities. Consistently, enhanced invasion and metastasis were demonstrated in ALKR1275Q/MYCN mice. These results collectively indicate that ALKR1275Q confers a malignant potential on neuronal cells that overexpress MYCN by impairing normal ECM/BM integrity and enhancing tumor growth and dissemination. Moreover, we found that crizotinib, an ALK inhibitor, almost completely inhibited the growth of ALKR1275Q/MYCN tumors in an allograft model. Our findings provided insights into the cooperative mechanism of the mutated ALK and overexpressed MYCN in the pathogenesis of NB and demonstrated the effectiveness of crizotinib on ALKR1275Q-positive tumors.

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Acknowledgements

We thank Yuki Sakai, Sawako Ogata and Rika Tai for animal care and the genotyping and molecular experiments. We also thank Dr Junji Takeda of Osaka University, RIKEN BioResource Center and Pfizer Inc. for providing us with KY1.1 ES cells, B6.Cg-Tg(CAG-Cre)CZ-MO2Osb mice (RBRC01828) and crizotinib, respectively. We also thank Dr William A Weiss of University of California San Francisco (UCSF) in US for allowing us to use the TH-MYCN Tg mice. We also thank Yumi Sugawara and Hiromi Kato at the University of Tokyo in Japan for LacZ staining.

Footnotes

ALKR1275Q mice have been deposited in RIKEN BioResource Center, (http://www.brc.riken.jp/inf/en/index.shtml, RBRC05496). RNA-seq data obtained in this study have been deposited under accession numbers DRX028665− DRX028672 in the DDBJ (DNA Data Bank of Japan) BioSample.

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Author notes

  1. T Ueda, Y Nakata and N Yamasaki: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Disease Model, Research Institute for Radiation Biology and Medicine, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima, Japan

    T Ueda, Y Nakata, N Yamasaki, K Ikeda, Y Sera & H Honda

  2. Department of Pathology, Tokyo Women’s Medical University, 8-1 Kawada-cho, Shinjuku-ku, Tokyo, Japan

    H Oda

  3. Department of Molecular Pathology, Hiroshima University Institute of Biomedical and Health Sciences, 1-2-3 Kasumi, Minami-ku, Hiroshima, Japan

    K Sentani & W Yasui

  4. Department of Molecular Oncology, Research Institute for Radiation Biology and Medicine, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima, Japan

    A Kanai

  5. Division of Gene Regulation, Institute for Advanced Medical Research, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-kui, Tokyo, Japan

    N Onishi & H Saya

  6. Health Care Center and Graduate School of Humanities and Sciences, Institute of Environmental Science for Human Life, Ochanomizu University, 2-1-1 Otsuka, Bunkyo-ku, Tokyo, Japan

    Z-i Honda

  7. Division for Health Service Promotion, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, Japan

    K Tanaka

  8. Department of Cardiovascular Medicine, Institute of Health Biosciences, The University of Tokushima Graduate School, Kuramotocho, Tokushima, Japan

    M Sata

  9. Department of Pathology, Graduate School of Medicine, Kyoto University, Yoshida-Konoe-cho, Sakyo-ku, Kyoto, Japan

    S Ogawa

  10. Department of Pediatrics, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, Japan

    J Takita

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  1. T Ueda
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Correspondence to H Honda.

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Ueda, T., Nakata, Y., Yamasaki, N. et al. ALKR1275Q perturbs extracellular matrix, enhances cell invasion and leads to the development of neuroblastoma in cooperation with MYCN. Oncogene 35, 4447–4458 (2016). https://doi.org/10.1038/onc.2015.519

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