P16INK4a Exerts an Anti-Inflammatory Effect through Accelerated IRAK1 Degradation in Macrophages

Abstract

Induction of cyclin-dependent kinase (CDK) inhibitor gene p16(INK4a) into the synovial tissues suppresses rheumatoid arthritis in animal models. In vitro studies have shown that the cell-cycle inhibitor p16(INK4a) also exerts anti-inflammatory effects on rheumatoid synovial fibroblasts (RSF) in CDK activity-dependent and -independent manners. The present study was conducted to discern how p16(INK4a) modulates macrophages, which are the major source of inflammatory cytokines in inflamed synovial tissues. We found that p16(INK4a) suppresses LPS-induced production of IL-6 but not of TNF-α from macrophages. This inhibition did not depend on CDK4/6 activity and was not observed in RSF. p16(INK4a) gene transfer accelerated LPS-triggered IL-1R-associated kinase 1 (IRAK1) degradation in macrophages but not in RSF. The degradation inhibited the AP-1 pathway without affecting the NF-κB pathway. Treatment with a proteosome inhibitor prevented the acceleration of IRAK1 degradation and downregulation of the AP-1 pathway. THP-1 macrophages with forced IRAK1 expression were resistant to the p16(INK4a)-induced IL-6 suppression. Senescent macrophages with physiological expression of p16(INK4a) upregulated IL-6 production when p16(INK4a) was targeted by specific small interfering RNA. These findings indicate that p16(INK4a) promotes ubiquitin-dependent IRAK1 degradation, impairs AP-1 activation, and suppresses IL-6 production. Thus, p16(INK4a) senescence gene upregulation inhibits inflammatory cytokine production in macrophages in a different way than in RSF.