NLRC5 expression in tumor cells is critical to activate adaptive and innate antitumor immune responses

Shukla, A.; Quenum, A. J. I.; Lucier, J.-F.; Cayarga, A. A.; Levesque, D.; Boisvert, F.-M.; Kufer, T. A.; Ramanathan, S.; Ilangumaran, S.

doi:10.64898/2025.12.19.695247

bioRxiv · 10.64898/2025.12.19.695247

NLRC5 expression in tumor cells is critical to activate adaptive and innate antitumor immune responses

Shukla, A.·Quenum, A. J. I.·Lucier, J.-F.·Cayarga, A. A.·Levesque, D.·Boisvert, F.-M.·Kufer, T. A.·Ramanathan, S.·Ilangumaran, S.

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Abstract

Tumors evade cytotoxic T lymphocyte (CTL)-mediated killing by downregulating MHC class-I, mainly resulting from the loss of its transcriptional activator NLRC5. Expressing full-length NLRC5 (NLRC5-FL) or a shorter NLRC5-CIITA fusion protein termed NLRC5 super-activator (NLRC5-SA) in cancer cells upregulates MHC-I expression and promotes antitumor immunity. To distinguish the role of NLRC5 expressed within tumor cells and antigen presenting cells, we studied B16-F10 melanoma expressing NLRC5-FL (B16-N-FL) or NLRC5-SA (B16-N-SA) in Nlrc5+/+ and Nlrc5-/- mice. Both tumors were efficiently controlled in both Nlrc5+/+ and Nlrc5-/- hosts with abundant immune cell infiltration, enriched for activated and differentiated CD8+ and CD4+ T cells, NK, NKT and iNKT cells. B16-N-FL and B16-N-SA tumors showed increased collagen deposition and vascularization, with upregulation of CCL4 and CXCL9 chemokine genes in B16-N-SA tumors. Depletion of either CD8+ T cells or NK1.1+ cells increased the growth of B16-N-FL and B16-N-SA tumors in Nlrc5+/+ mice, and that of B16-N-SA tumors in Nlrc5-/- hosts. Proteomes of B16-N-FL and B16-N-SA cells showed downmodulation of dominant tumor antigens and upregulation of ubiquitination and protein processing pathway proteins. Differentially expressed proteins shared between B16-N-FL and B16-N-SA cells showed enrichment in phagosome and autophagy pathways. We conclude that tumor cell-intrinsic NLRC5 expression is critical for the activation of adaptive and innate immune cells, and establishment of an immune-supportive tumor microenvironment to permit immune cell infiltration and their effector functions and achieve tumor control. NLRC5 expression in APCs is dispensable to mediate these effects. Delivering NLRC5-SA is a promising approach to restore antitumor immune responses in MHC-I-low immune evasive tumors. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=134 SRC="FIGDIR/small/695247v1_ufig1.gif" ALT="Figure 1"> View larger version (40K): org.highwire.dtl.DTLVardef@1bc4f51org.highwire.dtl.DTLVardef@1fb5b03org.highwire.dtl.DTLVardef@11448edorg.highwire.dtl.DTLVardef@bff26c_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Shukla, A., Quenum, A. J. I., Lucier, J.-F., Cayarga, A. A., Levesque, D., Boisvert, F.-M., Kufer, T. A., Ramanathan, S., Ilangumaran, S.. 2025-12-23. NLRC5 expression in tumor cells is critical to activate adaptive and innate antitumor immune responses. https://doi.org/10.64898/2025.12.19.695247

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