Abstract
Neoantigen-specific T cells specifically recognize tumor cells and are critical for cancer immunotherapies. However, the transcriptional program controlling the cell fate decisions by neoantigen-specific T cells is incompletely understood. Here, using joint single-cell transcriptome and TCR profiling, we mapped the clonal expansion and differentiation of neoantigen-specific CD8 (+) T cells in the tumor and draining lymph node in mouse prostate cancer. Compared to other antitumor CD8 (+) T cells and bystanders, neoantigen-specific CD8 (+) tumor-infiltrating lymphocytes (TILs) upregulated gene signatures of T cell activation and exhaustion. In the tumor draining lymph node, we identified TCF1 (+) TOX (-) T (SCM) , TCF1 (+) TOX (+) T (PEX) , and TCF1 (-) TOX (+) effector-like T (EX) subsets among neoantigen-specific CD8 (+) T cells. Clonal tracing analysis of neoantigen-specific CD8 (+) T cells revealed greater clonal expansion in divergent clones and less expansion in clones biased towards T (EX,) T (PEX) , or T (SCM) . The T (PEX) subset had greater clonal diversity and likely represented the root of neoantigen-specific CD8 (+) T cell differentiation, whereas highly clonally expanded effector-like T (EX) cells were positioned at the branch point where neoantigen-specific clones exited the lymph node and differentiated into T (EX) TILs. Notably, T (SCM) differentiation of neoantigen-specific CD8 (+) clones in the lymph node negatively correlated with exhaustion and clonal expansion of the same clones in the tumor. In addition, the gene signature of neoantigen-specific clones biased toward tumor infiltration relative to lymph node residence predicted a poorer response to immune checkpoint inhibitor. Together, we identified the transcriptional program that controls the cell fate choices by neoantigen-specific CD8 (+) T cells and correlates with clinical outcomes in cancer patients.