Inhibition of cellular communication network factor 1 (CCN1)-driven senescence slows down cartilage inflammaging and osteoarthritis

Objective: To explore the role of cellular communication network factor 1 (CCN1) in cartilage inflammaging and osteoarthritis (OA) pathogenesis in the isolated primary human chondrocytes in vitro, cartilage explants ex vivo, and a pre-clinical mice model.

Methods: Recombinant human CCN1 stimulation and small interfering RNA inhibition were conducted in human chondrocytes. The RNA was extracted to quantify catabolic targets and pro-inflammatory genes and the proteins were probed with specific antibodies. IL-1β and IL-6 were monitored by ELISA. IHC was performed to evaluate important hypertrophic hallmarks and catabolic markers. The effects of Tanshinone IIA on chondrocytes were investigated in both time-dependent and dose-dependent processes. Cartilage explants were cultured in growth medium and further treated with Tanshinone IIA. The intra-articular injection was performed in 13 months old C57BL/6J mice. Safranin O and fast green staining were performed to evaluate the histological change of cartilage followed by a semi-quantitative analysis using the OARSI scoring system.

Results: RNA and protein levels of CCN1 increased in an age-dependent manner compared to young donors. Increased CCN1 expression was also found in the damaged area compared to the non-lesion area which correlated with the advanced pathological change in human OA. The overexpression of CCN1 promoted chondrocytes senescence, while the down-regulation of CCN1 by small interfering RNA reduced CCN1 production and limited inflammation secretion suggesting that CCN1 was a possible novel target to intervene OA. Inhibition of CCN1 by using Tanshinone IIA could reduce SASP components in a dose- and time-dependent manner. Additionally, our data showed that Tanshinone IIA was able to preserve articular cartilage integrity, suppress CCN1 production, and inhibit SASP factors in human cartilage explants and in aged mice model.

Conclusion: This study showed that CCN1 signaling aggravated cartilage inflammaing and matrix degradation. Collectively, our findings showed new insight into repurposing Tanshinone IIA for slowing down OA advancement in human and mice by inhibiting the CCN1 axis.