Lu et al. developed a new approach to identify neoantigen-specific TCRs for use in adoptive T cell transfer. The process involves briefly co-culturing expanded tumor-infiltrating lymphocytes with autologous APCs that have been transfected with tandem minigenes containing patient-specific mutations or pulsed with mutant peptide, and using single-cell RNA sequencing to identify TCR sequences on activated T cells that express high levels of IFNγ and/or IL-2. A clinical trial to test this approach is being planned.

The adoptive transfer of neoantigen-reactive tumor-infiltrating lymphocytes (TILs) can result in tumor regression in patients with metastatic cancer. To improve the efficacy of adoptive T cell therapy targeting these tumor-specific mutations, we have proposed a new therapeutic strategy, which involves the genetic modification of autologous T cells with neoantigen-specific T cell receptors (TCRs) and the transfer of these modified T cells back to cancer patients. However, the current techniques to isolate neoantigen-specific TCRs are labor intensive, time consuming, and technically challenging, not suitable for clinical applications. To facilitate this process, a new approach was developed, which included the co-culture of TILs with tandem minigene (TMG)-transfected or peptide-pulsed autologous antigen-presenting cells (APCs) and the single-cell RNA sequencing (RNA-seq) analysis of T cells to identify paired TCR sequences associated with cells expressing high levels of interferon-gamma (IFN-gamma) and interleukin-2 (IL-2). Following this new approach, multiple TCRs were identified, synthesized, cloned into a retroviral vector, and then transduced into donor T cells. These transduced T cells were shown to specifically recognize the neoantigens presented by autologous APCs. In conclusion, this approach provides an efficient procedure to isolate neoantigen-specific TCRs for clinical applications, as well as for basic and translational research.

Author Info: (1) Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. Electronic address: yong-chen.lu@nih.gov. (2) Surgery Branch, National Cancer Institute, National Institutes

Author Info: (1) Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. Electronic address: yong-chen.lu@nih.gov. (2) Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. (3) Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. (4) Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. (5) Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. (6) Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. (7) Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. (8) Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. (9) Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. (10) Genetics Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. (11) Genome Analysis Unit, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. (12) Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. Electronic address: sar@nih.gov.

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