CFIm25-dependent alternative polyadenylation in AKT2 mRNA programs macrophage polarization

Macrophage polarization into M1 (pro-inflammatory) or M2 (anti-inflammatory) phenotypes is essential for immune responses and tissue homeostasis. While transcriptional and post-transcriptional mechanisms controlling this process have been characterized, the contribution of alternative mRNA polyadenylation (APA) to polarization requires clarification. Using human monocytic cell lines, we demonstrate that CFIm25, a key APA regulator, controls macrophage polarization states. CFIm25 overexpression enhances M1 characteristics, including nitric oxide production, CD80 expression, pro-inflammatory cytokine secretion, phagocytosis, migration, and cancer cell killing, while suppressing M2 traits. CFIm25 knockdown produces opposite effects. Mechanistically, CFIm25 promotes the proximal polyadenylation of AKT2 mRNA, generating shorter transcripts with enhanced stability and translational efficiency that increase Akt2 protein levels and amplify NF-κB signaling. Blocking the proximal site with antisense oligonucleotides reduces Akt2 expression and induces M2-like phenotypes. These findings establish APA as a critical regulator of macrophage polarization and identify the CFIm25-Akt2-NF-κB axis as a potential therapeutic target for modulating immune responses.