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Novel TENM3–ALK fusion is an alternate mechanism for ALK activation in neuroblastoma

Oncogene volume 41, pages 2789–2797 (2022)Cite this article

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Abstract

The identification of molecular events underlying the pathogenesis of neuroblastoma can likely result in improved clinical outcomes for this disease. In this study, a translocation within chromosome 2p and 4q was found to bring about the formation of an in-frame fusion gene that was composed of portions of the teneurin transmembrane protein 3 (TENM3, also known as ODZ3) gene and the anaplastic lymphoma kinase (ALK) gene in tumor cells from patients with neuroblastoma. Expression of the full length TENM3–ALK cDNA in NIH-3T3 cells led to the formation of a fusion protein that: (1) possesses constitutive tyrosine kinase activity, (2) induces strong activation of the downstream targets of extracellular signal-regulated kinase (ERK), protein kinase B (a.k.a. AKT), and signal transducer and activator of transcription 3 (STAT3), (3) provokes oncogenic transformation in NOD.Cg-PrkdcscidIl2rgtm1Sug/ShiJic mice, and (4) possesses sensitivity to ALK inhibitors in vitro and in vivo. Our findings demonstrated that patients with neuroblastoma may express a transforming fusion kinase, which is a promising candidate for a therapeutic target and a diagnostic molecular marker for neuroblastoma. The in-frame 5′ partner gene that fuses with ALK has not been reported previously in neuroblastoma. Our data provide novel biological insights into the mechanism of ALK activation due to translocation, with implications for neuroblastoma tumorigenesis, and could be useful as a vital marker for the accurate diagnosis of this type of neuroblastoma.

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Fig. 1: The translocations involving ALK and TENM3 breakpoints.
Fig. 2: The pathological analysis of TENM 3-ALK-rearranged NIH3T3 cells by ALK immunohistochemistry and immunofluorescence staining.
Fig. 3: Kinase activities of TENM3–ALK or ALK ex 1–11 deleted mutant and their downstream status.
Fig. 4: Effects of Lorlatinib or Crizotinib in cells expressing TENM3–ALK.
Fig. 5: In vivo transforming activity of TENM3–ALK inhibition of the growth of NIH3T3 cells expressing TENM3–ALK by a chemical inhibitor of ALK.

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Acknowledgements

The authors would like to thank M. Matsumura, K. Yin, and F. Saito for their excellent technical assistance, Dr. T. Kitamura (Division of Cellular Therapy/Division of Stem Cell Signaling, The Institute of Medical Sciences, The University of Tokyo, Japan) for the kind gift of expression vector pMYs-IRES-Neo. The authors also wish to express their appreciation to K. Chiba and H. Tanaka (The University of Tokyo) for the supercomputer. This work was supported by JSPS KAKENHI Grant Numbers JP19J11112, JP17H04224, JP18K19467, and JP20H00528; Project for Cancer Research and Therapeutic Evolution (P-CREATE; grant no. JP19cm0106509h9904), and Practical Research for Innovative Cancer Control (grant no. JP19ck0106468h001) from the Japan Agency for Medical Research and Development (AMED); Princess Takamatsu Cancer Research Fund. This research also used the computational resources of the K computer provided by the RIKEN Advanced Institute for Computational Science through the HPCI System Research project (hp140230, hp160219, and hp150232) (SM and SO).

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Authors and Affiliations

  1. Department of Pediatrics, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan

    Mitsuteru Hiwatari, Masafumi Seki, Takeshi Nagasawa, Aiko Sato-Otsubo, Motohiro Kato, Kentaro Watanabe & Masahiro Sekiguchi

  2. Department of Cell Therapy and Transplantation Medicine, The University of Tokyo Hospital, Tokyo, Japan

    Mitsuteru Hiwatari

  3. Department of Pediatrics, School of Medicine, Teikyo University, Tokyo, Japan

    Mitsuteru Hiwatari

  4. Department of Pediatrics, Kansai Medical University, Osaka, Japan

    Ryosuke Matsuno

  5. Department of Pathology and Tumor Biology, Graduate School of Medicine, Kyoto University, Yoshidakonoecho, Sakyo-ku, Kyoto, Japan

    Kenichi Yoshida & Seishi Ogawa

  6. Department of Pediatrics, Tokai University School of Medicine, Isehara, Kanagawa, Japan

    Shohei Yamamoto

  7. Medical and Dental Data Science Center, Tokyo Medical and Dental University, Tokyo, Japan

    Satoru Miyano

  8. Institute for the Advanced Study of Human Biology (WPI-ASHBi), Kyoto University, Yoshidakonoecho, Sakyo-ku, Kyoto, Japan

    Seishi Ogawa

  9. Department of Medicine, Center for Hematology and Regenerative Medicine, Karolinska Institute, Stockholm, Sweden

    Seishi Ogawa

  10. Department of Pediatrics, Kyoto University, Yoshidakonoecho, Sakyo-ku, Kyoto, Japan

    Junko Takita

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Contributions

MH and JT wrote the paper; MH, TN, and MS analyzed the data; MH performed the experiments; RM and SY analyzed the FISH data; KY, SM, and SO developed the bioinformatics pipelines; MK, ASO, MS, KW, and JT provided conceptual advice; and MH and JT designed the study. All authors read and approved the final paper.

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Correspondence to Mitsuteru Hiwatari.

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Hiwatari, M., Seki, M., Matsuno, R. et al. Novel TENM3–ALK fusion is an alternate mechanism for ALK activation in neuroblastoma. Oncogene 41, 2789–2797 (2022). https://doi.org/10.1038/s41388-022-02301-1

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