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Loss of cancer cell STAT1 improves response to radiation therapy and promotes T cell activation in head and neck squamous cell carcinoma

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Abstract

Resistance to radiation therapy (RT) remains an obstacle in HPV-negative head and neck squamous cell carcinomas (HNSCCs)—even with a combined RT-immunotherapy approach. Jak-Stat proteins have long been studied for both their immune regulatory role in the host immune response as well as their cancer cell signaling role in shaping the tumor microenvironment (TME). Here, we identify STAT1 as a mediator of radioresistance in HPV-negative preclinical mouse models of HNSCC, by which knockout of STAT1 in the cancer cell (STAT1 KO)—but not in the host—resulted in decreased tumor growth alongside increased immune activation. We show that RT increases STAT1/pSTAT1 expression, which may act as a marker of radioresistance. Whereas RT increased JAK-STAT and interferon (IFN) signaling, transcriptomic analysis revealed that STAT1 KO in the cancer cell resulted in decreased expression of IFN-associated genes of resistance. In vitro experiments showed that STAT1 KO increased T cell chemoattraction and decreased baseline growth. These results indicate that STAT1 may serve a tumor-promoting role in the cancer cell and will inform biomarker development and treatment regimens for HNSCC incorporating RT.

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Funding

Dr. Sana D. Karam is funded by the NIDCR/NCI (R01 DE028529-01, R01 DE028282-01, 1 P50 CA261605-01) and receives preclinical funding from Roche and clinical trial funding from AstraZeneca, both unrelated to this research. This study was also partly supported by the National Institutes of Health P30CA046934 Cancer Center Support Grant, including the Bioinformatics and Biostatistics Shared Resource, the Cancer Center Flow Cytometry Shared Resource, the Genomics and Microarray Shared Resource, and the Cell Technologies Shared Resource at the University of Colorado. This study was also partly supported by the Cancer League of Colorado.

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Authors and Affiliations

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Contributions

SDK and MWK designed the study and all experiments. MWK, TEB, SBu, and DN performed mouse studies. BVC performed irradiation. MWK, SBh, SC, DN, and AO performed in vitro experiments. AO performed cell proliferation assays and western blotting. MWK, LBD, and JG performed flow cytometry and analysis. MP and MWK performed scratch wound assays. Sbu and MWK performed boyden chamber assays and analysis. All authors advised throughout the course of the study. SDK and MWK wrote the manuscript. All authors discussed, revised, and approved the final manuscript.

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Correspondence to Sana D. Karam.

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The authors declare no potential conflicts of interest.

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The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Available from the corresponding author upon reasonable request.

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All listed authors have contributed significantly to the research described and have read and approved this manuscript for submission to Cancer Immunology, Immunotherapy.

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Précis

HNSCCs have responded poorly to current RT-immunotherapy combinations, and mechanisms of resistance remain elusive. Here, we provide compelling data for cancer cell-specific STAT1 directing resistance in immune -dependent and -independent behavior.

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Knitz, M.W., Darragh, L.B., Bickett, T.E. et al. Loss of cancer cell STAT1 improves response to radiation therapy and promotes T cell activation in head and neck squamous cell carcinoma. Cancer Immunol Immunother 71, 1049–1061 (2022). https://doi.org/10.1007/s00262-021-03059-3

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  • DOI: https://doi.org/10.1007/s00262-021-03059-3

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