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Timing of immune checkpoint blockade shapes anti-tumor immunity via a clock-dependent chemokine axis

Lichterman, J. N.; Srinivasan, T.; Wang, R.; Dende, C.; Zarek, C.; Ramirez, J. I.; Park, N.; Wang, H.; Li, W.; Kim, J.; Coughlin, L.; Poulides, N.; Sabaeifard, P.; Palmer, S. N.; Morrill, A. C.; Hassell, B.; Shelton, J. M.; Li, X. V.; Evers, B. M. M.; Zhan, X.; Takahashi, J. S.; Hooper, L. V.; Koh, A. Y.

2025-11-04 immunology
10.1101/2025.11.03.686293 bioRxiv
Show abstract

Circadian clocks regulate immunity, yet how they shape the tumor immune microenvironment and influence cancer immunotherapy remains unclear. Here, we show that tumor immune infiltration and immune checkpoint inhibitor efficacy vary by time of day in mice, driven by intrinsic clocks in dendritic cells and CD8+ T cells. Time-of-day modulates the abundance, spatial organization, and cytokine-chemokine production of tumor-infiltrating immune cells. Mechanistically, dendritic cell clocks control expression of Cx3cl1, driving recruitment of CX3CR1+CD8+ T cells and thereby reshaping the tumor immune microenvironment to enhance immunotherapy efficacy. Disruption of this axis abolishes time-of-day-dependent differences in treatment response. These findings identify a circadian mechanism of immune cell recruitment to tumors and provide mechanistic insight into clinical observations linking treatment timing to immunotherapy outcomes. One sentence summaryTime of day determines cancer immunotherapy efficacy through a circadian clock-dependent chemokine axis.

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