Exosomal Dvl3 promoted the aggressive phenotypic transformation of RA-FLS via wnt pathway

Abstract

Objective

This study was performed to explore the function and mechanism of Dvl3 in RA-FLS by exosome intervention.

Methods

The expression pattern of Dvl3 was examined by IHC, WB, and qPCR. Modified exosomes obtained from culturing supernatant of RA-FLS infected with Dvl3 over expression (OE) lentivirus were administrated to the target RA-FLS. The ability of survival, migration, and the production of inflammatory factor influenced by exosomal Dvl3 were detected by CKK8 kits, Tunel, migration test, qPCR, and enzyme-linked immunosorbent assay (ELISA) respectively; Flow cytometry analysis was conducted to explorer the inflammatory moderate role of exosomes on CD4+ T cells. The possible downstream pathways of Dvl3 were screened by qPCR and WB and verified by double luciferase reporter experiment.

Results

The expression level of Dvl3 was significantly increased in RA and CIA. Exosomes from the OE group could significantly promote cell proliferation activity, migration/invasion ability. The augment of TNF-α, IL-1β, IL-17, and IL-21 was observed in exosomal Dvl3-OE group. Th1 and Th17 cells polarisation and cytokines related were both enhanced by Exosomal Dvl3. Over expression of Dvl3 was accompanied by the significant increase of β-catenin and RhoA activities.

Conclusion

This study discovered the high expression of Dvl3 of exosomes derived from RA patients which may possessed the ability to promote phenotypic transformation of RA-FLS through Wnt pathway.

Keywords:

• RA-FLS
• Dvl3
• exosomes
• Wnt
• inflammation
• RhoA
• ROCK2

Ethics approval and consent to participate

The Institutional of Ethics Committee of the Shanghai Changhai Hospital reviewed and approved the study. All volunteers gave their written informed consent before participation.

All animal operations and handling have been approved by the Animal Care and Use Committee of Changhai Hospital.

Consent for publication

The authors give consent for publication on arthritis research & therapy.

Author contributions

T-WX and Z-DB designed the experiments, CN, Z-YJ and QY collected clinical specimens, CN and L-HR performed the morphological study. T-WX, CN and GJ performed the experiments. QY, F-XM and L-SW analysed and interpreted all of the data. T-WX and QY wrote the manuscript.

Disclosure statement

The authors declare there are no competing interests.

Data availability statement

All the data material related to the research are available.

Additional information

Funding

This work was supported by National Natural Science Foundation of China [81971484], and Natural Science Foundation of Shandong Province [ZR2020MH317].