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Published on: February 2, 2013
Direct identification of neoantigen-specific TCRs from tumor specimens by high-throughput single-cell sequencing
Yong-Chen Lu1,2, Zhili Zheng3, Frank J Lowery3
1Surgery Branch, National Institutes of Health, Bethesda, Maryland, USA YLu@uams.edu.
Background:
Recognition of neoantigens by T cells plays a major role in cancer immunotherapy. Identification of neoantigen-specific T-cell receptors (TCRs) has become a critical research tool for studying T cell-mediated responses after immunotherapy. In addition, neoantigen-specific TCRs can be used to modify the specificity of T cells for T cell-based therapies targeting tumor-specific mutations. Although several techniques have been developed to identify TCR sequences, these techniques still require a significant amount of labor, making them impractical in the clinical setting.
Methods:
Thanks to the availability of high-throughput single-cell sequencing, we developed a new process to isolate neoantigen-specific TCR sequences. This process included the isolation of tumor-infiltrating T cells from a tumor specimen and the stimulation of T cells by neoantigen-loaded dendritic cells, followed by single-cell sequencing for TCR and T-cell activation markers, interferon-γ and interleukin-2.
Results:
In this study, potential neoantigen-specific TCRs were isolated from three melanoma and three colorectal tumor specimens. These TCRs were then synthesized and transduced into autologous T cells, followed by testing the recognition of neoantigens. A total of 28 neoantigen-specific TCRs were identified by this process. If identical TCR sequences were detected from two or more single cells, this approach was highly reliable (100%, 19 out of 19 TCRs).
Conclusion:
This single-cell approach provides an efficient process to isolate antigen-specific TCRs for research and clinical applications.
Insights
Researchers developed an efficient single-cell sequencing method to isolate neoantigen-specific T-cell receptors (TCRs). This advance aids cancer immunotherapy research and clinical applications by enabling faster identification of critical TCR sequences.
Area of Science:
- Immunology
- Oncology
- Genomics
Background:
- T-cell receptor (TCR) recognition of neoantigens is crucial for cancer immunotherapy.
- Identifying neoantigen-specific TCRs is vital for studying T-cell responses and developing T-cell-based therapies.
- Current methods for TCR identification are labor-intensive and not clinically practical.
Purpose of the Study:
- To develop an efficient process for isolating neoantigen-specific TCR sequences using high-throughput single-cell sequencing.
- To enable faster and more practical identification of TCRs for research and clinical applications in cancer immunotherapy.
Main Methods:
- Isolated tumor-infiltrating T cells from melanoma and colorectal cancer specimens.
- Stimulated T cells with neoantigen-loaded dendritic cells.
- Performed single-cell sequencing to identify TCR sequences and T-cell activation markers (interferon-γ, interleukin-2).
Main Results:
- Successfully isolated 28 neoantigen-specific TCRs from six tumor specimens.
- Demonstrated high reliability of the method, with 100% accuracy for repeatedly detected TCR sequences.
- Synthesized and validated the identified TCRs in autologous T cells for neoantigen recognition.
Conclusions:
- The developed single-cell approach offers an efficient method for isolating antigen-specific TCRs.
- This technique has significant potential for both research and clinical applications in cancer immunotherapy.
- The process streamlines the identification of TCRs, overcoming limitations of previous labor-intensive methods.
