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De-differentiation in cultures of organoids from luminal-type breast cancer is restored by inhibition of NOTCH signaling

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Abstract

Estrogen receptor (ER) expression in breast cancer can change during progression and the treatment, but the mechanism has not been well studied. In this study, we successfully prepared organoids from samples obtained from 33 luminal-type breast cancer patients and studied their ER expression. The expression status was well maintained in primary organoids, whereas it decreased after passaging in most of the cases. In fact, the studied organoid lines were classified into those that retained a high level of ER expression (9%), those that completely lost it (9%), and those that repressed it to varying degrees (82%). In some cases, the ER expression was suddenly and drastically decreased after passaging. Marker protein immunohistochemistry revealed that after passaging, the differentiation status shifted from a luminal- to a basal-like status. Differentially expressed genes suggested the activation of NOTCH signaling in the passaged organoids, wherein a NOTCH inhibitor was able to substantially rescue the decreased ER expression and alter the differentiation status. Our findings suggest that the differentiation status of luminal-type cancer cells is quite flexible, and that by inhibiting the NOTCH signaling we can preserve the differentiation status of luminal-type breast cancer organoids.

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Data availability

The datasets generated and analyzed during the current study are available in DDBJ (accession number: DRR412393-DRR412396).

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Acknowledgements

We thank Mr. Hiromitsu Tazawa (Clinical Bio-resource Center of Kyoto University’s Hospital) for his administrative support and Mitsuaki Ishida for his support in the pathological analysis.

Funding

This research was supported by a collaboration grant from Kyoto University-KBBM (H.U., C.S., J.K., Y.M., M.M., T.S., M.I,) and the Takeda Science Foundation (M.I.).

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Author notes

  1. Jumpei Kondo

    Present address: Division of Health Sciences, Department of Molecular Biology and Clinical Investigation, Graduate School of Medicine, Osaka University, Osaka, Japan

Authors and Affiliations

  1. Department of Clinical Bio-Resource Research and Development, Graduate School of Medicine, Kyoto University, 46-29, Shimoadachi-cho, Sakyo-ku, Kyoto, 606-8304, Japan

    Hiroyuki Uematsu, Chieko Saito, Jumpei Kondo, Kunishige Onuma, Roberto Coppo & Masahiro Inoue

  2. KBBM Inc, 46-29, Shimoadachi-cho, Sakyo-ku, Kyoto, 606-8304, Japan

    Hiroyuki Uematsu & Chieko Saito

  3. Department of Therapeutic Oncology, Kyoto University Graduate School of Medicine, 54 Shogoin Kawahara-cho, Sakyo-ku, Kyoto, 606-8507, Japan

    Yukiko Mori & Manabu Muto

  4. Department of Breast Surgery, Kansai Medical University, Hirakata, Osaka, 573-1191, Japan

    Yuichiro Kikawa, Manami Tada & Tomoharu Sugie

  5. Department of Clinical Bio-Resource Research and Development, Graduate School of Medicine, Kyoto University, Med-Pharm Collaboration Building 503, Shimoadachi-cho 46-29, Sakyou-ku, Kyoto, 606-8304, Japan

    Masahiro Inoue

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Contributions

MM, YM, TS, and MI designed the study. HU and CS performed the experiments. JK and RO supervised the experiments. YK, MT, and TS acquired the clinical samples and data. HU, RO, and MI conducted the data analysis. HU and MI wrote the manuscript. HU, JK, KO, RO, and TS reviewed and revised the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Masahiro Inoue.

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Conflict of interest

J.K., K.O., and M.I. are members of the Department of Clinical Bio-resource Research and Development at Kyoto University, which is sponsored by KBBM, Inc. H.U. and C.S. are employees of KBBM Inc. The other authors declare no conflict of interest. This work was supported in part by the collaboration grant from KBBM-Kyoto University.

Ethics approval and consent to participate

This study was approved by the institutional ethics committees of Kyoto University (R2220, R2444) and Kansai Medical University (2019245, 2021090). Written informed consent was obtained from participants. This study was performed in accordance with the Declaration of Helsinki.

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Uematsu, H., Saito, C., Kondo, J. et al. De-differentiation in cultures of organoids from luminal-type breast cancer is restored by inhibition of NOTCH signaling. Human Cell 36, 2099–2112 (2023). https://doi.org/10.1007/s13577-023-00975-7

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  • DOI: https://doi.org/10.1007/s13577-023-00975-7

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