Phenotypic signatures of circulating neoantigen-reactive CD8+T cells in patients with metastatic cancers

Circulating T cells from peripheral blood (PBL) can provide a rich and noninvasive source for antitumor T cells. By single-cell transcriptomic profiling of 36 neoantigen-specific T cell clones from 6 metastatic cancer patients, we report the transcriptional and cell surface signatures of antitumor PBL-derived CD8+ T cells (NeoTCRPBL). Comparison of tumor-infiltrating lymphocyte (TIL)-and PBL-neoantigen-specific T cells re-vealed that NeoTCRPBL T cells are low in frequency and display less-dysfunctional memory phenotypes rela-tive to their TIL counterparts. Analysis of 100 antitumor TCR clonotypes indicates that most NeoTCRPBL pop-ulations target the same neoantigens as TILs. However, NeoTCRPBL TCR repertoire is only partially shared with TIL. Prediction and testing of NeoTCRPBL signature-derived TCRs from PBL of 6 prospective patients demonstrate high enrichment of clonotypes targeting tumor mutations, a viral oncogene, and patient-derived tumor. Thus, the NeoTCRPBL signature provides an alternative source for identifying antitumor T cells from PBL of cancer patients, enabling immune monitoring and immunotherapies.