RT Journal Article
SR Electronic
T1 Direct identification of neoantigen-specific TCRs from tumor specimens by high-throughput single-cell sequencing
JF Journal for ImmunoTherapy of Cancer
JO J Immunother Cancer
FD BMJ Publishing Group Ltd
SP e002595
DO 10.1136/jitc-2021-002595
VO 9
IS 7
A1 Yong-Chen Lu
A1 Zhili Zheng
A1 Frank J Lowery
A1 Jared J Gartner
A1 Todd D Prickett
A1 Paul F Robbins
A1 Steven A Rosenberg
YR 2021
UL http://jitc.bmj.com/content/9/7/e002595.abstract
AB 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.Data are available upon reasonable request. Next-generation sequencing data will be available upon request, following publication of this manuscript.