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Translational Therapeutics

Development of CAR-T cells specifically targeting cancer stem cell antigen DNAJB8 against solid tumours

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Abstract

Background

As therapy for solid tumours, various tumour antigens have been selected as targets, but CAR-T cells targeting these antigens have shown limited efficacy, in contrast to the effectiveness of CAR-T cells targeting haematological malignancies. In a previous report, we identified a cancer-testis antigen, DNAJB8. DNAJB8 plays a major role in tumorigenicity in cancer stem-like cells/cancer-initiating cells (CSCs/CICs). Here, we report a DNAJB8-reactive CAR yielding anti-tumour effects against renal cell carcinoma (RCC) and osteosarcoma.

Methods

We constructed a second-generation chimeric antigen receptor (CAR) against HLA-A*24:02/DNAJB8-derived peptide (DNAJB_143) complex (B10 CAR). The reactivity of B10-CAR T cells against T2-A24 cells pulsed with the cognate peptide and an RCC and osteosarcoma cell lines were quantified. The effects of adoptive cell transfer (ACT) therapy were assessed using in vivo xenografted mice models.

Results

B10 CAR-T cells recognised DNAJB8_143-pulsed T2-A24 cells and HLA-A*24:02(+)/DNAJB8(+) renal cell carcinoma and osteosarcoma cell lines. Moreover, ACT using B10 CAR-T cells showed anti-tumour effects against RCC and osteosarcoma cells.

Conclusion

B10 CAR-T cells could show specific cytotoxicity against RCC and osteosarcoma cells in vitro and in vivo. B10 CAR-T cells targeting the CSC/CIC antigen DNAJB8 might be a candidate immunotherapy for carcinoma and sarcoma.

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Fig. 1: Construction of B10 CAR-T cells recognising the HLA-A*24:02/DNAJB8_143 complex.
Fig. 2: Reactivity of B10 CAR-T cells against peptide-pulsed antigen-presenting cells.
Fig. 3: Reactivity of B10 CAR-T cells against various cell lines.
Fig. 4: B10 CAR-T cells exert anti-tumour effects in a xenografted mouse model.

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Data availability

All data generated or analysed during this study are included in this article and the supplementary files.

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Acknowledgements

We thank Ms. Asuka Akamatsu for technical assistance.

Funding

This work was supported by grants from the Japan Society for the Promotion of Science (KAKENHI 20H03807 to T Tsukahara and 17H01540 to T Torigoe), the Princess Takamatsu Cancer Research Fund (19-25125 to T Tsukahara), the Kobayashi Foundation for Cancer Research (15-9 to T Tsukahara), and AMED (21cm0106309h0006 to T Torigoe and 22ym0126801j0001 to T Tsukahara).

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Authors and Affiliations

  1. Department of Pathology, Sapporo Medical University, School of Medicine, South-1, West-17, Chuo-ku, Sapporo, Hokkaido, 060-8556, Japan

    Yuto Watanabe, Tomohide Tsukahara, Kenji Murata, Shuto Hamada, Terufumi Kubo, Takayuki Kanaseki, Yoshihiko Hirohashi & Toshihiko Torigoe

  2. Department of Orthopedic Surgery, Sapporo Medical University School of Medicine, South-1, West-16, Chuo-ku, Sapporo, Hokkaido, 060-8543, Japan

    Yuto Watanabe, Shuto Hamada, Makoto Emori, Atsushi Teramoto & Toshihiko Yamashita

  3. Department of Diagnostic Pathology, Tokyo Medical University Hachioji Medical Center, 1163 Tatemachi, Hachioji, Tokyo, 193-0998, Japan

    Munehide Nakatsugawa

Authors
  1. Yuto Watanabe
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  2. Tomohide Tsukahara
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  7. Yoshihiko Hirohashi
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  10. Munehide Nakatsugawa
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  12. Toshihiko Torigoe
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Contributions

T Tsukahara designed the studies, YW, T Tsukahara, KM and SH acquired data and prepared samples, ME provided materials, T Kubo., T Kanaseki, YH and MN interpreted data, YW and T Tsukahara wrote the manuscript, and AT, TY and T Torigoe supervised the study.

Corresponding author

Correspondence to Tomohide Tsukahara.

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Competing interests

The authors declare no competing interests.

Ethics approval and consent to participate

This study was performed by the guidelines established by the Declaration of Helsinki and was approved by the Ethics Committee of Sapporo Medical University. Healthy donors provided informed consent for the use of their blood samples in our research.

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Consent for publication was obtained.

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Watanabe, Y., Tsukahara, T., Murata, K. et al. Development of CAR-T cells specifically targeting cancer stem cell antigen DNAJB8 against solid tumours. Br J Cancer 128, 886–895 (2023). https://doi.org/10.1038/s41416-022-02100-1

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