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Chronic myeloproliferative neoplasms

Mutant calreticulin interacts with MPL in the secretion pathway for activation on the cell surface

Leukemia volume 34, pages 499–509 (2020)Cite this article

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Abstract

Studies have shown that mutant calreticulin (CALR) constitutively activates the thrombopoietin (TPO) receptor MPL and thus plays a causal role in the development of myeloproliferative neoplasms (MPNs). To further elucidate the molecular mechanism by which mutant CALR promotes MPN development, we studied the subcellular localization of mutant CALR and its importance for the oncogenic properties of mutant CALR. Here, mutant CALR accumulated in the Golgi apparatus, and its entrance into the secretion pathway and capacity to interact with N-glycan were required for its oncogenic capacity via the constitutive activation of MPL. Mutant CALR-dependent MPL activation was resistant to blockade of intracellular protein trafficking, suggesting that MPL is activated before reaching the cell surface. However, removal of MPL from the cell surface with trypsin shut down downstream activation, implying that the surface localization of MPL is required for mutant CALR-dependent activation. Furthermore, we found that mutant CALR and MPL interact on the cell surface. Based on these findings, we propose a model in which mutant CALR induces MPL activation on the cell surface to promote MPN development.

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Acknowledgements

This work was funded in part by the MEXT-Supported Program for the Strategic Research Foundation at Private Universities; MEXT’s Promotion Plan for the Platform of Human Resource Development for Cancer Project; the JSPS KAKENHI Grants #15K15368, #16K09859, #17K16195, #17H04211, #18K08372, #18K16126, # 18K16127, #18K16098, and #19K08848; grants from the Takeda Science Foundation, the SENSHIN Medical Research Foundation and the Japan Leukemia Research Fund; and personal research funds from Haruhito Hosoya, Yuki Ishibe, Yoshitsugu Katayose, Mikio Kato, Masae Kunitama, Kumie Kurita, Masahiro Nakamura, Kaori Taki, Tetsuya Takahashi, Kanji Shishido, and Hiroaki Yamakawa. The funders had no role in the preparation of the manuscript. We are grateful to Shing Leng Chan for her critical reading of the manuscript and to Soichiro Kakuta and Kazuhito Naka for their productive suggestions. We would like to thank other members of the Department of Hematology for supporting this study. We would also like to acknowledge the Laboratory of Molecular and Biochemical Research, the Laboratory of Morphological Analysis and Imaging, and the Division of Cell Biology in the Research Support Center of the Juntendo University Graduate School of Medicine.

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Author notes

  1. These authors contributed equally: Nami Masubuchi, Marito Araki

Authors and Affiliations

  1. Department of Hematology, Juntendo University Graduate School of Medicine, Tokyo, Japan

    Nami Masubuchi, Yinjie Yang, Erina Hayashi, Misa Imai, Yoko Edahiro, Yumi Hironaka, Yoshihisa Mizukami, Yoshihiko Kihara, Hiraku Takei, Mai Nudejima & Norio Komatsu

  2. Research Institute for Disease of Old Age, Juntendo University Graduate School of Medicine, Tokyo, Japan

    Nami Masubuchi

  3. Department of Transfusion Medicine and Stem Cell Regulation, Juntendo University Graduate School of Medicine, Tokyo, Japan

    Marito Araki & Akimichi Ohsaka

  4. Leading Center for the Development and Research of Cancer Medicine, Juntendo University Graduate School of Medicine, Tokyo, Japan

    Misa Imai & Yoshihiko Kihara

  5. Center for Genomic and Regenerative Medicine, Juntendo University Graduate School of Medicine, Tokyo, Japan

    Yoshihisa Mizukami

  6. Department of Cell Biology and Neuroscience, Juntendo University Graduate School of Medicine, Tokyo, Japan

    Masato Koike

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  1. Nami Masubuchi
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Contributions

Conceived and designed the experiments: NM, MA, and NK. Performed the experiments: NM, MA, YY, EH, MI, YE, YH, YM, YK, HT and MN. Analyzed the data: NM, MA, YY, EH, MI and YE. Contributed reagents/materials/analysis tools: MK, AO and NK. Wrote the paper: NM, MA and NK.

Corresponding author

Correspondence to Norio Komatsu.

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Masubuchi, N., Araki, M., Yang, Y. et al. Mutant calreticulin interacts with MPL in the secretion pathway for activation on the cell surface. Leukemia 34, 499–509 (2020). https://doi.org/10.1038/s41375-019-0564-z

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