Aging-associated Increase of GATA4 levels in Articular Cartilage is Linked to Impaired Regenerative Capacity of Chondrocytes and Osteoarthritis

Although the causal association between aging and osteoarthritis (OA) has been documented, our understanding of the underlying mechanism remains incomplete. To define the regulatory molecules governing chondrocyte aging, we performed transcriptomic analysis of young and old human chondrocytes from healthy donors. The data predicted that GATA binding protein 4 (GATA4) may play a key role in mediating the difference between young and old chondrocytes. Results from immunostaining and western blot showed significantly higher GATA4 levels in old human or mouse chondrocytes when compared to young cells. Moreover, overexpressing GATA4 in young chondrocytes remarkably reduced their cartilage-forming capacity in vitro and induced the upregulation of proinflammatory cytokines. Conversely, suppressing GATA4 expression in old chondrocytes, through either siRNA or a small-molecule inhibitor NSC140905, increased the production of aggrecan and collagen type II, and also decreased levels of matrix-degrading enzymes. In OA mice, induced by surgical destabilization of the medial meniscus, intraarticular injection of lentiviral vectors carrying mouse Gata4 resulted in a higher OA severity, synovial inflammation, and pain level when compared to control vectors. Mechanistically, we found that overexpressing GATA4 significantly increased the phosphorylation of SMAD1/5. Our work demonstrates that the aging-associated increase of GATA4 in chondrocytes plays a vital role in OA progression, which may also serve as a target to reduce osteoarthritis in the older population. Overall design: In this study, to understand key regulators that contribute to an “aged” state in chondrocytes prior to OA onset, we first compared the transcriptome of young and old human chondrocytes isolated from healthy donors without known joint diseases and employed unbiased analysis to define the key regulatory molecules that mediate chondrocyte aging. Assessment of the chondrocyte genome demonstrated the upregulation of several factors in aged chondrocytes, and transcription factor GATA binding protein 4 (GATA4) specifically drew our attention since it is associated with DNA damage and cellular senescence. Upregulation of GATA4 in OA chondrocytes was also reported.We thus hypothesized that the increased GATA4 level contributes to chondrocyte aging and accelerated OA progression upon injuries. We first tested the hypothesis by increasing or suppressing GATA4 expression in healthy chondrocytes and examining their cartilage-forming capacity. We also assessed the role of GATA4 in vivo by injecting lentiviral vectors carrying control or mouse Gata4 genes into the knee joints of OA mice induced by surgically created destabilization of the medial meniscus (DMM). Lastly, a mechanistic study was conducted to explore how GATA4 impacts chondrocyte phenotypes and functions.