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Divergent role of CD8 T cells with distinct metabolic phenotypes during curative radio-immunotherapy in hot versus cold tumors

Heaton, A. R.; Burkard, N. J.; Gao, A.; Hoefges, A.; Feils, A. S.; Burkard, S. K.; Felder, M. A.; Tsarovsky, N. W.; Spiegelman, D. V.; Lublin, G. M.; Hampton, A. A.; D'Amato, A.; Dinh, H. Q.; Rakhmilevich, A. L.; Erbe, A. K.; Sondel, P. M.; Skala, M. C.

2026-01-05 immunology
10.1101/2025.10.24.684274 bioRxiv
Show abstract

Immunotherapy has potential for impactful cancer cures by empowering patients own immune cells. We developed a radio-immunotherapy regimen that can cure large immunologically hot and cold murine tumors. Here, we explored the divergent role of CD8 T cells during this radio-immunotherapy in contrasting hot colon carcinoma versus cold melanoma. We introduced an immunocompetent mouse model with mCherry-expressing CD8 T cells to provide cell tracking in vivo. We investigated single-cell function, metabolism, and gene expression temporal changes using flow cytometry, in vivo multiphoton imaging, single-cell RNA sequencing, and multiplexed immunofluorescence to determine the underlying mechanisms. We found that in contrast to the hot colon carcinoma model, CD8 T cells from the cold melanoma model do not drive tumor cures, despite getting activated, possibly due to a static oxidative metabolism and exhausted phenotype plus down regulation of tumor MHC-I expression. These findings have implications for improving immunotherapy response in immunologically cold cancers.

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