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Updated: Jul 18, 2025
Identification of Mediators of T-cell Receptor Signaling via the Screening of Chemical Inhibitor Libraries
Published on: January 22, 2019
A systematic safety pipeline for selection of T-cell receptors to enter clinical use
Zsofia Foldvari1,2, Cathrine Knetter1,2, Weiwen Yang1,2
1Department of Cancer Immunology, Institute for Cancer Research, Oslo University Hospital Radiumhospitalet, Oslo, Norway.
Abstract:
Cancer immunotherapy using T cell receptor-engineered T cells (TCR-Ts) represents a promising treatment option. However, technologies for pre-clinical safety assessment are incomplete or inaccessible to most laboratories. Here, TCR-T off-target reactivity was assessed in five steps: (1) Mapping target amino acids necessary for TCR-T recognition, followed by (2) a computational search for, and (3) reactivity screening against, candidate cross-reactive peptides in the human proteome. Natural processing and presentation of recognized peptides was evaluated using (4) short mRNAs, and (5) full-length proteins. TCR-Ts were screened for recognition of unintended HLA alleles, and as proxy for off-target reactivity in vivo, a syngeneic, HLA-A*02:01-transgenic mouse model was used. Validation demonstrated importance of studying recognition of full-length candidate off-targets, and that the clinically applied 1G4 TCR has a hitherto unknown reactivity to unintended HLA alleles, relevant for patient selection. This widely applicable strategy should facilitate evaluation of candidate therapeutic TCRs and inform clinical decision-making.
Insights
Assessing T cell receptor-engineered T cell (TCR-T) off-target reactivity is crucial for cancer immunotherapy safety. A new strategy evaluates peptide recognition, HLA allele cross-reactivity, and in vivo models to ensure therapeutic efficacy and patient safety.
Area of Science:
- Immunology
- Oncology
- Biotechnology
Background:
- T cell receptor-engineered T cells (TCR-Ts) show promise for cancer immunotherapy.
- Pre-clinical safety assessment technologies for TCR-Ts are limited.
- Off-target reactivity poses a significant safety concern for TCR-T therapies.
Purpose of the Study:
- To develop and validate a comprehensive strategy for assessing TCR-T off-target reactivity.
- To identify unintended peptide recognition and HLA allele cross-reactivity.
- To evaluate the clinical relevance of identified off-target reactivities.
Main Methods:
- Mapping TCR-T target amino acids for recognition.
- Computational screening and in vitro reactivity testing of cross-reactive peptides.
- Evaluation of natural peptide processing and presentation using mRNA and full-length proteins.
- Screening for unintended HLA allele recognition and in vivo validation in a mouse model.
Main Results:
- A novel five-step strategy for assessing TCR-T off-target reactivity was established.
- The study identified previously unknown reactivity of the 1G4 TCR to unintended HLA alleles.
- Validation confirmed the importance of studying full-length protein recognition for accurate safety assessment.
Conclusions:
- The developed strategy facilitates robust pre-clinical evaluation of therapeutic TCRs.
- Understanding TCR-T off-target reactivity is essential for patient selection and clinical decision-making.
- This approach enhances the safety profile of TCR-T cancer immunotherapies.
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