Discovering tumor-reactive T-cell receptors through single-cell sequencing of tumor-infiltrating lymphocytes

Abstract

We evaluated the utility of single-cell sequencing of tumor-infiltrating lymphocytes (TIL) for tumor-reactive T-cell receptor (TCR) discovery. Using the MC38 cell line as our tumor model in mice, we show that expression of exogenous TCRs via mRNA electroporation in human T cells provides an easy and quick path to validating tumor-specific candidate TCRs. We detail the identification and validation of four novel MC38-reactive mouse TCRs with varying levels of reactivity to the target cells. Validating our process, one of the MC38 TCRs is specific against a previously reported neoantigen (ASMTNMELM in the Adpgk gene). Consideration of these methodologies may aid in the development of rapid TCR-based therapies for the treatment of cancer and human disease.Abstract Figure

Lactose blocks intercellular spreading of Galectin-1 from cancer cells to T-cells and activates tumor immunological control

“…The largest TCR clone, CN1 (CDR3: TRA;TRB=CAIDPYNQGKLIF; CASSPGQGYAEQFF), against a specific neoantigen (ASMTNMELM in the Adpgk gene) of MC38 cells 48 increased from 2.3% to 34.2% in all CD3 + , 2.3% to 32.4% in CD8_Tex, 0.03% to 0.8% in CD8_Tpex, and 0.05% to 0.9% in CD8_EffectorMemory after lactose treatment (Fig. S24e-f).…”