Early dynamics of a neutrophil interferon program drive immunotherapy resistance and limit durable anti-tumor immunity

Gao, Y.; Anning, E.; Wang, L.; CHENG, Y.; Wu, L.; Wang, J.; Liu, F.; Xu, Z.; Hao, X.; Liu, J.; Yu, L.; Ding, Y.; Zhang, W.; Edwards, D. G.; Wu, Y.-H.; Chan, H. L.; Hoffman, D.; Wang, S.; Lee, T. D.; Dominguez, L. B.; Peled, N.; Li, X.; Chen, X.; Rivas, C. H.; Zhao, N.; Calderon, S. J.; Pedroza, D. A.; Smith, A.; Guan, N.; Liu, Z.; Rosen, J.; Zhang, X.

doi:10.64898/2026.09.30.755684

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bioRxiv · 10.64898/2026.09.30.755684

Early dynamics of a neutrophil interferon program drive immunotherapy resistance and limit durable anti-tumor immunity

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Abstract

Despite the remarkable success of immune checkpoint blockade (ICB), durable clinical benefit remains limited to a subset of patients, highlighting the need for predictive biomarkers and new therapeutic targets. Here, we investigated the early immune dynamics associated with anti-PD1 responsiveness using longitudinal single-cell transcriptomics, spatial transcriptomics, and neutrophil-specific genetic perturbations in preclinical models of triple-negative breast cancer. We identified a conserved neutrophil interferon-stimulated gene (ISG) program that emerges before overt tumor divergence and predicts resistance to anti-PD1 therapy. Successful treatment was associated with an early reduction of IFN-responsive neutrophils in both blood and tumor tissues, whereas persistent neutrophil IFN signaling correlated with therapeutic resistance. Mechanistically, genetic disruption of either type I or type II interferon signaling in neutrophils enhanced anti-PD1 efficacy and promoted durable anti-tumor immune memory. Spatial analyses further revealed an inverse association between IFN-responsive neutrophils and tertiary lymphoid structure-enriched regions during effective therapy. Together, these findings identify IFN-responsive neutrophils as early determinants of immunotherapy outcome and establish dynamic neutrophil interferon programs as candidate blood-based biomarkers and therapeutic targets for cancer immunotherapy.

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Gao, Y., Anning, E., Wang, L., CHENG, Y., Wu, L., Wang, J., Liu, F., Xu, Z., Hao, X., Liu, J., Yu, L., Ding, Y., Zhang, W., Edwards, D. G., Wu, Y.-H., Chan, H. L., Hoffman, D., Wang, S., Lee, T. D., Dominguez, L. B., Peled, N., Li, X., Chen, X., Rivas, C. H., Zhao, N., Calderon, S. J., Pedroza, D. A., Smith, A., Guan, N., Liu, Z., Rosen, J., Zhang, X.. 2026-10-01. Early dynamics of a neutrophil interferon program drive immunotherapy resistance and limit durable anti-tumor immunity. https://doi.org/10.64898/2026.09.30.755684

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