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GRIK2 is a target for bladder cancer stem-like cell-targeting immunotherapy


Recent studies have revealed that treatment-resistant cancer stem-like cells (CSCs)/cancer-initiating cells (CICs) can be targeted by cytotoxic T lymphocytes (CTLs). CTLs recognize antigenic peptides derived from tumor-associated antigens; thus, the identification of tumor-associated antigens expressed by CSCs/CICs is essential. Human leucocyte antigen (HLA) ligandome analysis using mass spectrometry enables the analysis of naturally expressed antigenic peptides; however, HLA ligandome analysis requires a large number of cells and is challenging for CSCs/CICs. In this study, we established a novel bladder CSC/CIC model from a bladder cancer cell line (UM-UC-3 cells) using an ALDEFLUOR assay. CSCs/CICs were isolated as aldehyde dehydrogenase (ALDH)-high cells and several ALDHhigh clone cells were established. ALDHhigh clone cells were enriched with CSCs/CICs by sphere formation and tumorigenicity in immunodeficient mice. HLA ligandome analysis and cap analysis of gene expression using ALDHhigh clone cells revealed a distinctive antigenic peptide repertoire in bladder CSCs/CICs, and we found that a glutamate receptor, ionotropic, kainite 2 (GRIK2)-derived antigenic peptide (LMYDAVHVV) was specifically expressed by CSCs/CICs. A GRIK2 peptide-specific CTL clone recognized GRIK2-overexpressing UM-UC-3 cells and ALDHhigh clone cells, indicating that GRIK2 peptide can be a novel target for bladder CSC/CIC-targeting immunotherapy.

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The authors are grateful to DNAFORM for performing CAGE. This work was supported by the Japan Society for the Promotion of Science, KAKENHI for T. Torigoe (17H01540) and Y. Hirohashi (20H03460). This work was also supported by the Japan Agency for Medical Research and Development, the Project for Cancer Research and Therapeutic Evolution (P-CREATE) for T. Torigoe (16770510) and T. Kanaseki (20cm0106352h0002), and the Japan Science and Technology Agency, CREST (JPMJCR15G3) for S. Hashimoto.

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All authors contributed to the study conception and design. Material preparation, data collection, and analysis were performed by HM, YH, SY, JY, AM, SH, ST, KH, TA, TK, TT, TK, NS, and TT. The first draft of the manuscript was written by HM, YH, NS, and TT, and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Yoshihiko Hirohashi or Toshihiko Torigoe.

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Miyata, H., Hirohashi, Y., Yamada, S. et al. GRIK2 is a target for bladder cancer stem-like cell-targeting immunotherapy. Cancer Immunol Immunother 71, 795–806 (2022).

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  • Bladder cancer
  • Cancer stem cell
  • GRIK2
  • Antigen
  • Cytotoxic T lymphocytes