A post-transcriptional regulatory checkpoint controls the response of tumor-infiltrating cytotoxic CD4+ T cells to immunotherapy

Vila de Mucha, M.; Hlond, J.; Costoya, C.; Lau, C.-I.; Sledzinska, A.; Uddin, I.; Navarrete, M.; Shah, M.; Mastrokalos, G.; Karagianni, D.; Bell, S. E.; Nattress, C.; Beattie, G.; Tape, C. J.; Turner, M.; Quezada, S. A.; Jenner, R. G.

doi:10.1101/2025.09.09.675154

bioRxiv · 10.1101/2025.09.09.675154

A post-transcriptional regulatory checkpoint controls the response of tumor-infiltrating cytotoxic CD4+ T cells to immunotherapy

Vila de Mucha, M.·Hlond, J.·Costoya, C.·Lau, C.-I.·Sledzinska, A.·Uddin, I.·Navarrete, M.·Shah, M.·Mastrokalos, G.·Karagianni, D.·Bell, S. E.·Nattress, C.·Beattie, G.·Tape, C. J.·Turner, M.·Quezada, S. A.·Jenner, R. G.

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Abstract

Acquisition of cytotoxic activity in CD4+ T cells (TCTX) can promote potent anti-tumor activity thus holding promise as a therapeutic approach. However, how this activity is regulated remains poorly understood. Here, we demonstrate that tumor-infiltrating CD4+ TCTX activity is restrained by a post-transcriptional regulatory checkpoint. In untreated tumors, CD4+ TCTX exist in a poised state, characterized by abundant Gzmb mRNA but limited Granzyme B (GzmB) protein. Differentiation into poised TCTX is regulated by the Blimp-1-Bcl6 axis and requires type-I interferon signaling. Treatment with anti-CTLA-4 or anti-LAG-3 plus anti-PD-1 removed the block to GzmB protein production by repressing expression of the post-transcriptional regulator Zfp36l1. Constitutive Zfp36l1 expression abrogated the effects of anti-CTLA-4 while deletion of Zfp36l1 and its paralog Zfp36 triggered GzmB protein production and promoted tumor control. These data identify ZFP36/ZFP36L1 as a key post-transcriptional regulatory checkpoint of CD4+ TCTX activity and a potential immunotherapy target in cancer.

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Vila de Mucha, M., Hlond, J., Costoya, C., Lau, C.-I., Sledzinska, A., Uddin, I., Navarrete, M., Shah, M., Mastrokalos, G., Karagianni, D., Bell, S. E., Nattress, C., Beattie, G., Tape, C. J., Turner, M., Quezada, S. A., Jenner, R. G.. 2025-09-12. A post-transcriptional regulatory checkpoint controls the response of tumor-infiltrating cytotoxic CD4+ T cells to immunotherapy. https://doi.org/10.1101/2025.09.09.675154

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