Related Experiment Video
Updated: Apr 30, 2026
A Nonviral Approach to Generate Transient Chimeric Antigen Receptor T Cells Using mRNA for Cancer Immunotherapy
Published on: February 21, 2025
A platform for parallel TCR cloning and testing enables anti-neoantigen tumor immunotherapy
Alexander M Rowe1, Smriti Chaurasia1, Wenzhong Wei1,2
1Department of Immunology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
Abstract:
Tumor-infiltrating CD8 cells recognize neoantigens created by tumor-specific mutations. Nonetheless, even after checkpoint inhibitor therapy, most patients' tumors progress. A deeper understanding of antitumor responses could facilitate development of better therapies. To enable such studies, we applied TCXpress, a high throughput platform that clones fully expressible TCRs from single cells into retroviral or lentiviral vectors without sequencing or gene synthesis, to study TCRs from CD8 cells infiltrating mouse MC38 tumors. We expressed cloned TCRs in reporter cells and interrogated TCR specificity by coculturing them with B6WT3 cells transduced with tandem minigenes encoding predicted neoantigens. We isolated TCRs reactive against epitopes from mutant Rpl18, Adpgk, Psmd2, and Zc3h7b along with self-reactive TCRs that recognized normal B6 and MC38 cells. Importantly, we successfully treated MC38-bearing mice with T cells transduced with anti-Rpl18 TCRs. These results establish a system that could be used to study many types of T cell responses and validate a therapeutic approach that could be tested in the clinic.
Insights
Researchers developed TCXpress to identify T cell receptors (TCRs) targeting cancer neoantigens. This platform enabled the discovery of TCRs that successfully treated mice with MC38 tumors, validating a potential new cancer therapy.
Area of Science:
- Immunology
- Oncology
- Biotechnology
Background:
- CD8 T cells recognize tumor neoantigens, but most patients do not respond to current immunotherapies.
- Understanding anti-tumor T cell responses is crucial for developing improved cancer treatments.
Purpose of the Study:
- To develop a high-throughput platform (TCXpress) for cloning and studying T cell receptors (TCRs) from tumor-infiltrating CD8 cells.
- To identify specific TCRs targeting neoantigens in mouse MC38 tumors and evaluate their therapeutic potential.
Main Methods:
- Applied TCXpress to clone TCRs from CD8 cells infiltrating MC38 tumors.
- Expressed cloned TCRs in reporter cells and tested specificity against neoantigen-presenting cells.
- Administered T cells engineered with anti-Rpl18 TCRs to MC38-bearing mice.
Main Results:
- Identified TCRs reactive against neoantigens from Rpl18, Adpgk, Psmd2, and Zc3h7b mutations.
- Isolated self-reactive TCRs that recognized normal B6 and MC38 cells.
- Successfully treated MC38-bearing mice using T cells transduced with anti-Rpl18 TCRs.
Conclusions:
- TCXpress provides a powerful system for studying T cell responses against tumor neoantigens.
- TCR-engineered T cells targeting specific neoantigens represent a viable therapeutic strategy for cancer treatment.
