C/EBPβ-induced alternative splicing of RCAN1 generates a potent TCR-T target in mesenchymal glioblastoma
Glioblastoma (GBM) is an aggressive brain tumor with limited treatment options and a dismal prognosis. While immunotherapy has shown promise in treating some solid tumors, the treatment of GBM has been mostly unsuccessful because of a lack of targetable tumor antigens and high tumor heterogeneity. Here, we report RCAN1-4 as a novel tumor antigen derived from alternative splicing induced by the transcription factor C/EBPβ. Both C/EBPβ and RCAN1-4 are highly expressed in GBM and glioma stem cells as mesenchymal subtype hallmarks. We report an immunogenic HLA-A24-specific splicing junction epitope within exon 4 and exon 5 that is unique to RCAN1-4. This epitope was validated for its ability to stimulate T cell responses in HLA-A24<sup>+</sup> donors and GBM patients, leading us to identify RCAN1-4-reactive T cell receptors (TCRs) for the construction of TCR-engineered T cells (TCR-T cells). Functional studies of TCR-Ts demonstrated the in vitro and in vivo killing of RCAN1-4<sup>pos</sup> GBM tumor cells, highlighting its potential as an immunotherapeutic target in mesenchymal GBM.
Authors
- Poulomi Nath (ORCID: https://orcid.org/0000-0002-1063-5427)
- Yuanyuan Han (ORCID: https://orcid.org/0000-0001-7043-5390)
- Thomas G. Forsthuber
- Lora H. Rigatti (ORCID: https://orcid.org/0000-0003-3921-0593)
- Megan Mantica (ORCID: https://orcid.org/0000-0002-6107-6696)
- Muzi Li
- Megan A. Mahlke
- Rebecca A. Raphael (ORCID: https://orcid.org/0009-0004-6869-7382)
- Gary Kohanbash
- Pascal Olivier Zinn (ORCID: https://orcid.org/0000-0003-0091-5887)
- Yihao Zhang (ORCID: https://orcid.org/0000-0001-7023-0221)
- Ambika P. Jaswal (ORCID: https://orcid.org/0000-0002-1929-0082)
- Zujian Xiong (ORCID: https://orcid.org/0000-0003-2943-1931)
- Bin Qin (ORCID: https://orcid.org/0000-0002-9180-0550)
- Xuejun Li
- Bhuvitha Chagantipati
- Yapeng Chao
- Sydney A Jackson
- Amelia Stepniak
- Chloe R Kuminkoski
- Itay Raphael
- Baoli Hu
- Wei Zhang
- Chaim T. Sneiderman
- Qinglin Kong
- Yael Nechemia-Arbely
- Ian F. Pollack
- Lance Schwegman
- Zeynep Dulkadir
- Sameer Agnihotri
Institutions
- Central South University (CN)
- University of Pittsburgh (US)
- Hebrew University of Jerusalem (IL)
- Anhui Medical University (CN)
- Cornell University (US)
- Zhengzhou University (CN)
- UPMC Hillman Cancer Center (US)
- Neurological Surgery (US)
- Xiangya Hospital Central South University (CN)
- The University of Texas at San Antonio (US)
Publication Details
- Journal
- Cellular and Molecular Immunology
- Published
- 2025-12-23
- DOI
- https://doi.org/10.1038/s41423-025-01360-0
- Citations
- 2
- Primary Topic
- Signaling Pathways in Disease
- Type
- article
- Field-Weighted Citation Impact
- 1.61
Funders
- University of Pittsburgh
- Central South University
- National Institutes of Health
- University of California, San Francisco
- Center for Research Computing, University of Pittsburgh
- National Cancer Institute
- Children's Hospital of Pittsburgh