Abstract
Osteoarthritis (OA) is an age-related disease characterized by cartilage degeneration. TNFR1-associated death domain protein (TRADD) is a key upstream molecule of TNF-α signals but its role in OA pathogenesis is unknown. This study aimed to verify that whether inhibition of TRADD could protect against chondrocyte necroptosis and OA, and further elucidate the underlying mechanism. We demonstrated that TNF-α-related OA-like phenotypes including inflammation response, extracellular matrix degradation, apoptosis, and necroptosis in chondrocytes were inhibited by TRADD deficiency. Furthermore, TRADD interacted with TRAF2 and knockdown of TRADD suppressed the activation of RIPK1-TAK1-NF-κB signals and restored impaired autophagy. ICCB-19, the selective inhibitor of TRADD, also attenuated necroptosis in chondrocytes. Mechanismly, ICCB-19 blocked the phosphorylation of TAK1-NF-κB signals and restored impaired autophagy, whereas inhibiting autophagic process with 3-Methyladenine compromised these effects of ICCB-19. The in vivo study showed that the intra-articular injection of ICCB-19 rescued the expression of collagen alpha-1(II) chain and LC3, and mitigated the cartilage degeneration of OA mice. This study demonstrates that TRADD mediates TNF-α-induced necroptosis and OA-like phenotypes of chondrocytes and suggests that ICCB-19 suppresses chondrocyte damage and cartilage degeneration by inhibiting TNF-α-TRADD-mediated signals and dysregulation of autophagy in chondrocytes. ICCB-19 may serve as an important option for OA therapy.
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Introduction
Osteoarthritis (OA) is a common disease that causes physical pain and even disability in patients. This disease affects the majority of older people in the world and brings huge economic burden to society [1, 2]. A variety of risk factors have been identified for OA [3], but the clear pathogenesis and effective treatment remain to be further studied. The pathological changes of OA are characterized by chondrocyte death and cartilage destruction, synovitis, and osteophyte formation [4]. Chondrocytes and their secreted extracellular matrix (ECM) are main components of articular cartilage [5]. Chondrocytes maintain physiological structure of articular cartilage and improve joint function by well-regulating the balance of anabolism and catabolism of ECM [4]. Tumor necrosis factor-α (TNF-α) is a key inflammatory mediator in OA progression and one of the leading cytokines giving rise to the imbalance of matrix synthesis and decomposition [6, 7]. Therefore, it is important to maintain homeostasis of chondrocytes by protecting chondrocytes against death and degradation of ECM under the pathological conditions.
TNFR1-associated death domain protein (TRADD), is an adaptor molecule for the signaling downstream of tumor necrosis factor receptor 1 (TNFR1) [47]. The sections were incubated with anti-LC3, MMP13, P-RIPK3, and COL2A1 antibodies to observe expression changes of these markers.
Statistical analysis
Graph Pad Prism 8.0 software was used for statistical analysis. Results were revealed as mean ± standard deviation (SD). Student T test (unpaired, two-tailed) was used for statistical analysis between the two groups. Comparisons between more than two groups were performed by ANOVA and Tukey test. P values <0.05 was statistically significant. N = 3 means the in vitro experiments were performed at least three biological replicates.
Data availability
Full images of western blotting had been included in Supplementary Materials. Other data are available from the corresponding authors for reasonable grounds.
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Funding
This work was supported by the National Natural Science Foundation of China (82172498), the Project of Health Commission of Hubei Province (WJ2021M123), and the Knowledge Innovation Project of Wuhan (2022020801010442).
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WTZ Conception, design and obtaining of funding. FJG Conception and obtaining of funding. KS Data acquisition, analysis and drafting of the manuscript. ZG Data acquisition, analysis. JMZ Conception and design. LCH Data acquisition. SL: Technical support. FL Data acquisition and revision of the manuscript. GCW Analysis and interpretation of the data. JTX Technical support. XZ Technical support. All authors read and approved the final manuscript.
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Sun, K., Guo, Z., Zhang, J. et al. Inhibition of TRADD ameliorates chondrocyte necroptosis and osteoarthritis by blocking RIPK1-TAK1 pathway and restoring autophagy. Cell Death Discov. 9, 109 (2023). https://doi.org/10.1038/s41420-023-01406-0
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DOI: https://doi.org/10.1038/s41420-023-01406-0
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