Discovery of tumor-reactive T cell receptors by massively parallel library synthesis and screening
Ziva Moravec1, Yue Zhao2, Rhianne Voogd1
1Department of Molecular Oncology and Immunology, Netherlands Cancer Institute, Amsterdam, The Netherlands.
Abstract:
T cell receptor (TCR) gene therapy is a potent form of cellular immunotherapy in which patient T cells are genetically engineered to express TCRs with defined tumor reactivity. However, the isolation of therapeutic TCRs is complicated by both the general scarcity of tumor-specific T cells among patient T cell repertoires and the patient-specific nature of T cell epitopes expressed on tumors. Here we describe a high-throughput, personalized TCR discovery pipeline that enables the assembly of complex synthetic TCR libraries in a one-pot reaction, followed by pooled expression in reporter T cells and functional genetic screening against patient-derived tumor or antigen-presenting cells. We applied the method to screen thousands of tumor-infiltrating lymphocyte (TIL)-derived TCRs from multiple patients and identified dozens of CD4+ and CD8+ T-cell-derived TCRs with potent tumor reactivity, including TCRs that recognized patient-specific neoantigens.
Insights
This study introduces a new method for discovering tumor-reactive T cell receptors (TCRs) for personalized cancer immunotherapy. The pipeline efficiently identifies potent TCRs, including those targeting patient-specific neoantigens, advancing TCR gene therapy.
Area of Science:
- Immunology
- Oncology
- Biotechnology
Background:
- T cell receptor (TCR) gene therapy offers potent cellular immunotherapy by engineering T cells for tumor recognition.
- Isolating effective TCRs is challenging due to the rarity of tumor-specific T cells and unique patient tumor epitopes.
Purpose of the Study:
- To develop a high-throughput, personalized pipeline for discovering therapeutic TCRs.
- To identify novel TCRs with potent reactivity against patient-derived tumors and neoantigens.
Main Methods:
- Assembly of complex synthetic TCR libraries in a one-pot reaction.
- Pooled expression of TCRs in reporter T cells.
- Functional genetic screening against patient tumor or antigen-presenting cells.
Main Results:
- Screened thousands of tumor-infiltrating lymphocyte (TIL)-derived TCRs from multiple patients.
- Identified dozens of CD4+ and CD8+ T-cell-derived TCRs with significant tumor reactivity.
- Discovered TCRs capable of recognizing patient-specific neoantigens.
Conclusions:
- The developed pipeline enables efficient discovery of personalized TCRs for cancer immunotherapy.
- This approach facilitates the identification of TCRs targeting unique tumor antigens, enhancing therapeutic potential.
