FOXA2 inhibits the TLR4/NF-κB signaling pathway and alleviates inflammatory activation of macrophages in rheumatoid arthritis by repressing LY96 transcription.

Background: Rheumatoid arthritis (RA) remains a devastating autoimmune disease characterized by joint damage, inflammation, and disability. This study investigates the function of lymphocyte antigen 96 (LY96) in the inflammatory response in RA and explores its regulatory mechanism.

Methods: A mouse model of RA was developed using type II collagen, and the LY96 expression in the ankle joint tissue was determined. Upstream regulators targeting LY96 were investigated using bioinformatics, followed by chromatin immunoprecipitation and luciferase reporter assays for validation. Gain- or loss-of-functions of LY96 and forkhead box A2 (FOXA2) were performed to analyze their roles in arthritis score, pathological changes, and inflammatory responses in mice. The effects of FOXA2 and LY96 on pro-inflammatory activation of macrophages were additionally investigated in vitro using a mouse RAW264.7 macrophage model with lipopolysaccharide treatment.

Results: LY96 mRNA and protein (MD-2) levels were increased in the RA mice. Knockdown of LY96 alleviated arthritis severity, joint deformities, inflammation, and cartilage destruction in mice. In vitro, the LY96 knockdown reduced the pro-inflammatory activation of RAW264.7 macrophages by inhibiting the TLR4/NF-κB inflammatory signaling transduction. FOXA2 was identified as a transcriptional repressor of LP96 poorly expressed in RA. Overexpression of FOXA2 similarly alleviated inflammation and reduced M1-type macrophages in vivo and in vitro. However, these changes were reversed by the additional LY96 upregulation.

Conclusion: This study suggests that FOXA2 represses LY96 transcription to inhibit the TLR4/NF-κB signaling transduction, thus reducing pro-inflammatory activation of macrophages in the context of RA.