B4GALT1 drives osteoarthritis progression by stabilizing IL-1R1 through N-linked glycosylation

Osteoarthritis (OA) is a prevalent degenerative joint disease with an unclear molecular pathogenesis. B4GALT1 has been implicated in various pathological processes, but its role and mechanism in OA remain largely unexplored.

The expression of B4GALT1 was examined in OA clinical samples and experimental OA models. In vivo OA models were established by destabilization of the medial meniscus (DMM) in mice, while in vitro OA models were generated by lipopolysaccharide (LPS) stimulation of human chondrocytes. CCK-8, flow cytometry, qRT-PCR, and Western blot were employed to investigate the role of B4GALT1 in chondrocyte viability, apoptosis, and inflammatory response. The B4GALT1-IL-1R1 protein interaction was analyzed by Co-IP, and N-glycosylation was assessed using PNGase F treatment and site-directed mutagenesis. OA progression was further evaluated by SO/FG staining, H&E, and X-ray following intra-articular AAV-mediated B4GALT1 knockdown in DMM mice.

B4GALT1 was upregulated in OA samples. Functionally, B4GALT1 overexpression exacerbated LPS-induced chondrocyte apoptosis, decline in cell viability, and inflammatory cytokine imbalance (increased TNF-α and IL-1β, decreased IL-13), while its knockdown reversed these effects. Mechanistically, B4GALT1 directly interacted with IL-1R1 and promoted its N-linked glycosylation, specifically at the N193 site, thereby enhancing IL-1R1 protein stability. In vivo, AAV-mediated knockdown of B4GALT1 attenuated DMM-induced cartilage degeneration, joint space narrowing, and inflammatory responses, concomitant with reduced IL-1R1 protein levels.

B4GALT1 is upregulated in OA and promotes disease progression by stabilizing IL-1R1 via N-glycosylation at the N193 site. Targeting B4GALT1 may represent a promising therapeutic strategy for the treatment of OA.