Function and transcriptional profiles comparison between tumor-infitlrating NK cell (tuNK) and splenic NK cell (spNK) [RNA-seq]

Natural killer (NK) cells, a type of potent cytotoxic lymphocytes, are particularly promising for the treatment of cancers that lose or downregulate major histocompatibility complex class I (MHC-I) expression to evade T cell-mediated immunotherapy. However, the hostile and immune suppressive tumor microenvironment (TME) greatly hinders the function of tumor-infiltrating NK cells limiting the therapeutic efficacy against solid tumors. Here, we show that a fusion protein of interleukin-21 (IL-21-Fc), as a direct in vivo intervention, can safely and effectively reprogram NK cell metabolism and enhance their effector function in the TME. Our research demonstrates that combining IL-21-Fc with IL-15 superagonist (IL-15SA) or NK cell transfer leads to the eradication of MHC-I-deficient tumors and confers durable protection in syngeneic and xenograft tumor models. Mechanistically, we uncover that IL-21-Fc enhances NK cell effector function by upregulating glycolysis in a lactate dehydrogenase A (LDHA)-dependent manner. These findings not only underscore the considerable potential of IL-21-Fc as an in vivo therapeutic intervention to bolster NK cell-based immunotherapy, but also unveil an innovative strategy of metabolic reprogramming for NK cell rejuvenation within tumors. Overall design: To explore and compare the transcriptomic profiles of the intratumoral and splenic NK cells, in addition to assess the function of NK cells within the TME. We established a subcutaneous (s.c.) MHC-I-deficient tumor model by inoculating BALB/c mice with CT26_ß2m-/- murine colorectal carcinoma cells and performed bulk RNA-seq of sorted tissue-infiltrating NK cells.