CD39⁺ tissue-resident memory CD8⁺ T cells with a clonal overlap across compartments mediate antitumor immunity in breast cancer

Despite being a standard treatment option in breast cancer, immune checkpoint inhibitors (ICIs) are only efficacious for a subset of patients. To gain a better understanding of the antitumor immune response in breast cancer, we examined the heterogeneity of CD8⁺ T cells in tumors, metastatic lymph nodes (mLNs), and peripheral blood from patients with early breast cancer (n = 131). Among tissue-resident memory CD8⁺ T (T_RM) cells, including virus- and tumor-specific CD8⁺ T cells, CD39 expression was observed in a tumor-specific and exhausted subpopulation in both tumors and mLNs. CD39⁺ T_RM cells from tumors and mLNs exhibited a phenotypic similarity and clonally overlapped with each other. Moreover, tumor or mLN CD39⁺ T_RM cells clonally overlapped with CD39⁻ T_RM and non-T_RM cells in the same compartment, implying a tissue-specific differentiation process. These inter-subpopulationally overlapping CD39⁺ T_RM clonotypes were frequently detected among effector memory CD8⁺ T cells in peripheral blood, suggesting a systemic clonal overlap. CD39⁺ T_RM cell enrichment was heterogeneous among molecular subtypes of breast cancer, which is associated with the different role of antitumor immune responses in each subtype. In vitro blockade of PD-1 and/or CTLA-4 effectively restored proliferation of CD39⁺ T_RM cells and enhanced cytokine production by CD8⁺ T cells from tumors or mLNs, particularly in the presence of CD39⁺ T_RM enrichment. This suggests that CD39⁺ T_RM cells have a capacity for functional restoration upon ICI treatment. Thus, our study indicates that CD39⁺ T_RM cells with a clonal overlap across compartments are key players in antitumor immunity in breast cancer.