Study on the association between Mettl3-modified Piezo1 in CD4+ T cells and inflammatory response in ankylosing spondylitis.

OBJECTIVE

To explore the association between Mettl3-modified Piezo1 in CD4+ T cells and the inflammatory response in ankylosing spondylitis (AS).

METHODS

Peripheral venous blood and epidemiological datas were collected from 53 AS patients and 53 healthy controls. Enzyme-linked immunosorbent assay (ELISA) was employed to measure the levels of inflammatory cytokines. Real-time quantitative PCR (qRT-PCR) was utilized to detect the transcriptional levels of Mettl3 and Piezo1. A Mettl3-knockdown human Jurkat T cell model was established using siRNA technology. Western blot was performed to detect the protein levels of genes. MeRIP-PCR was used to validate the modification effect of Mettl3 on Piezo1.

RESULTS

The transcriptional level of Mettl3 was downregulated, while that of Piezo1 was upregulated in CD4+ T cells from AS patients, showing a negative correlation between them. ROC curve analysis indicated that both Mettl3 and Piezo1 possessed diagnostic capabilities, with the AUC for Mettl3 being 0.661 (95% CI: 0.555-0.767) and for Piezo1 being 0.938 (95% CI: 0.890-0.987). Subgroup analysis revealed that the Piezo1 mRNA level was significantly lower in the group using biologic agents compared to the group not using them. Cellular experimental results demonstrated that si-Mettl3 could elevate the protein level of Piezo1 in Jurkat T cells, accompanied by the increase of TNF-α. MeRIP-PCR further confirmed the role of Mettl3 in the m6A methylation modification of Piezo1.

CONCLUSIONS

The inhibition of Mett13 in CD4+ T cells in ankylosing spondylitis enhances the expression level of Piezo1 protein through m6A methylation modification, promoting the development of inflammatory responses in ankylosing spondylitis.