Related Experiment Video
Updated: Aug 27, 2025
Non-Viral Engineering of Primary Human T Cells via Homology-Mediated End-Joining Targeted Integration of Large DNA Templates
Published on: May 9, 2025
High-throughput T cell receptor engineering by functional screening identifies candidates with enhanced potency and
Rodrigo Vazquez-Lombardi1, Johanna S Jung2, Fabrice S Schlatter2
1Department of Biosystems Science and Engineering, ETH Zurich, Mattenstrasse 26, 4058 Basel, Switzerland; Engimmune Therapeutics AG, Hegenheimermattweg 167A, 4123 Allschwil, Switzerland.
Abstract:
A major challenge in adoptive T cell immunotherapy is the discovery of natural T cell receptors (TCRs) with high activity and specificity to tumor antigens. Engineering synthetic TCRs for increased tumor antigen recognition is complicated by the risk of introducing cross-reactivity and by the poor correlation that can exist between binding affinity and activity of TCRs in response to antigen (peptide-MHC). Here, we developed TCR-Engine, a method combining genome editing, computational design, and deep sequencing to engineer the functional activity and specificity of TCRs on the surface of a human T cell line at high throughput. We applied TCR-Engine to successfully engineer synthetic TCRs for increased potency and specificity to a clinically relevant tumor-associated antigen (MAGE-A3) and validated their translational potential through multiple in vitro and in vivo assessments of safety and efficacy. Thus, TCR-Engine represents a valuable technology for engineering of safe and potent synthetic TCRs for immunotherapy applications.
Insights
A new method, TCR-Engine, enhances T cell receptor (TCR) engineering for adoptive T cell immunotherapy. It improves TCR potency and specificity against tumor antigens, offering a promising tool for cancer treatment.
Area of Science:
- Immunology
- Biotechnology
- Cancer Research
Background:
- Adoptive T cell immunotherapy relies on T cell receptors (TCRs) with high tumor antigen specificity and activity.
- Engineering synthetic TCRs faces challenges like cross-reactivity and poor binding affinity-to-activity correlation.
Purpose of the Study:
- To develop a high-throughput method for engineering functional activity and specificity of TCRs.
- To create safer and more potent synthetic TCRs for cancer immunotherapy.
Main Methods:
- Developed TCR-Engine, integrating genome editing, computational design, and deep sequencing.
- Engineered TCRs on a human T cell line for high-throughput functional assessment.
- Applied TCR-Engine to target the MAGE-A3 tumor antigen.
Main Results:
- Successfully engineered synthetic TCRs with enhanced potency and specificity for MAGE-A3.
- Demonstrated translational potential through in vitro and in vivo safety and efficacy assessments.
- Validated TCR-Engine as a valuable technology for TCR engineering.
Conclusions:
- TCR-Engine enables high-throughput engineering of synthetic TCRs with improved functional activity and specificity.
- The engineered TCRs show promise for safe and potent application in adoptive T cell immunotherapy.
- This technology advances the development of effective cancer immunotherapies.
