The Role of WNT5A-mediated proteomic and phosphoproteomic regulatory networks in rheumatoid arthritis

Abstract

Objective

This study aims to investigate the role of the WNT5A signaling pathway in rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLS) and uncover the impact of WNT5A on cellular function and signal transduction through proteomic and phosphoproteomic analyses.

Methods

MH7A cells were treated with recombinant WNT5A (rhWNT5A), and differential expression proteins (DEPs) and phosphoproteins (DEPPs) were identified through proteomic and phosphoproteomic analyses. Data were further analyzed via volcano plots, heatmaps, enrichment analysis, and protein-protein interaction (PPI) networks to identify key biological processes and signaling pathways regulated by WNT5A.

Results

Significant changes in the expression of numerous DEPs and DEPPs were observed following rhWNT5A treatment, including proteins closely related to lipid metabolism, cell migration, inflammation, and cell proliferation. PPI network analysis revealed that key regulatory proteins, such as HNRNPA1, RANBP2, BCLAF1, NPM1, and SMARCA4, occupy central positions in the network. Enrichment analysis indicated that WNT5A influences essential signaling pathways, such as AMPK, mTOR, VEGFA-VEGFR2, Notch, and endoplasmic reticulum stress, regulating cellular energy metabolism, inflammatory response, and cytoskeletal remodeling. Kinase activity analysis identified significant changes in kinases such as CDK1, CSNK2A1, EEF2K, AURKA, and AURKB, which were further integrated into the kinase-substrate regulatory network.

Conclusion

This study demonstrates that WNT5A significantly influences the biological functions and inflammatory responses of RA-FLS by regulating key biological processes and signaling pathways. The integrated proteomic and phosphoproteomic analyses provide insights into the potential mechanisms and regulatory networks of WNT5A in RA, suggesting its application as a potential therapeutic target.