CaMKII induces an autophagy-dependent anabolic response in Articular Chondrocytes

Abstract

AbstractObjectiveThe objective of this study was to elucidate the role of Calcium calmodulin-dependent Kinase II (CaMKII) in articular chondrocytes and its involvement in osteoarthritis (OA) pathogenesis. By performing gain and loss of function experiments, the research aimed to determine how CaMKII modulates chondrocyte metabolism, anabolic and catabolic processes, hypertrophic differentiation, and autophagy within the articular cartilage.DesignArticular cartilage was harvested from patients undergoing joint replacement surgery for OA, and adult human articular chondrocytes (AHACs) were isolated and cultured. Recombinant adenoviruses were used to overexpress a constitutively active form of CaMKIIγ (AdCaMKII) or inhibit CaMKII activity (AdAIP). Various assays, including RT-PCR analysis, alcian blue staining of Micromass cultures, immunofluorescence, and Western blotting, were performed to assess the effects of CaMKII modulation on chondrocyte function.ResultsOverexpression of activated CaMKIIγ promoted anabolism, evidenced by increased expression of SOX9, COL2A1, and ACAN, and decreased MMP-13 levels. It also enhanced proteoglycan content in AHAC micromass cultures. Furthermore, CaMKII counteracted the catabolic effects of IL-1β and preserved proteoglycan content. We also observed decreased chondrocyte proliferation and increased synthesis of hypertrophic marker Type X Collagen. CaMKII activation was found to induce autophagy, as indicated by increased phosphorylation of Beclin1 and decreased p62 expression. The anabolic effects of CaMKII were dependent on autophagy, as inhibition of autophagy with Bafilomycin prevented the CaMKII-induced increase in glycosaminoglycan content.ConclusionsCaMKII plays a significant role in modulating chondrocyte metabolism and maintaining cartilage homeostasis. It promotes anabolic processes, counteracts catabolic stimuli, and induces autophagy in articular chondrocytes. However, it also promotes hypertrophic differentiation, highlighting the complexity of CaMKII-mediated signalling in cartilage. Understanding these pathways could lead to new therapeutic strategies that leverage CaMKII’s anabolic potential while mitigating its pro-degenerative effects.