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Mol Ther. 2018 Feb 7;26(2):379-389. doi: 10.1016/j.ymthe.2017.10.018. Epub 2017 Oct 28.

An Efficient Single-Cell RNA-Seq Approach to Identify Neoantigen-Specific T Cell Receptors.

Author information

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
Genetics Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
4
Genome Analysis Unit, 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. Electronic address: sar@nih.gov.

Abstract

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-γ (IFN-γ) 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.

KEYWORDS:

cancer immunotherapy; gene therapy; single cell

PMID:
29174843
PMCID:
PMC5835023
DOI:
10.1016/j.ymthe.2017.10.018
[Indexed for MEDLINE]
Free PMC Article

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