AF6 orchestrates macrophage polarization via JAK2-STAT3 signaling and promotes intestinal regeneration through stem cell proliferation

Macrophages play a central role in regulating intestinal inflammation, resolution, and tissue repair. however, the upstream regulators that govern their polarization during colitis remain poorly defined. Here, using a DSS-induced colitis model, we identify the scaffold protein AF6 was as a key regulator that promoting pro-inflammatory macrophage polarization and exacerbating colitis. Mechanistically, AF6 expression promoted JAK-STAT3 complex formation. In turn, its deletion in macrophages resulted in impaired JAK-STAT3 signaling pathway and a shift away from pro-inflammatory M1 polarization. Furthermore, the AF6-deficient macrophages were determined to also promote epithelial regeneration through enhanced IL-10 production. Thus, our study defines a therapeutically amenable AF6-JAK2-STAT3 axis that controls macrophage-driven pathogenesis in colitis, revealing a novel strategy to break the cycle of inflammation and impaired repair.