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miR-34c-5p is a novel regulator of T cell differentiation that targets FOXP3

Noronha-Estima, C.; Gaspar-Luta, R.; Araujo, T. F.; Ferreira-Fernandes, C.; Poiares-Oliveira, G.; Paulo-Pedro, M.; Marques, M. P.; Sousa, A. E.; Gama-Carvalho, M.

2025-07-26 immunology
10.1101/2025.07.25.665590 bioRxiv
Show abstract

MicroRNAs regulate key genes and pathways essential for T cell differentiation and function; however, many remain poorly characterized. We previously identified miR-34c-5p as a T cell receptor-inducible microRNA in naive CD4 T cells, but its role in immune regulation remained largely unexplored. In this study, we mapped miR-34c expression across multiple sort-purified CD4 T cell subsets and found that its induction is restricted to FOXP3 cells, both natural regulatory T cells (Tregs) and cells undergoing inducible Treg (iTreg) differentiation trajectory. miR-34c expression correlates with high FOXP3 levels and is absent in conventional memory effector subsets. Functional studies using miR-34c antagomiRs reveal that miR-34c limits iTreg differentiation by restraining FOXP3 expression. Mechanistically, we show that FOXP3 activates miR-34c transcription through direct binding to its promoter, while miR-34c targets the FOXP3 3'UTR, establishing a negative feedback loop. Together, our findings identify miR-34c as a FOXP3-responsive miRNA that fine-tunes iTreg development via post-transcriptional repression of FOXP3, uncovering a novel layer of regulatory control in T cell responses.

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