MicroRNA-21 from bone marrow mesenchymal stem cell-derived extracellular vesicles targets TET1 to suppress KLF4 and alleviate rheumatoid arthritis

Abstract

Background: Accumulating evidence has demonstrated that bone marrow mesenchymal stem cells (BMSCs)-derived extracellular vesicles (EVs) can be used effectively to transfer drugs and biomolecules to target lesions. Meanwhile, BMSCs have been reported to be beneficial in the treatment of rheumatoid arthritis (RA). In this study, we employ gain- and loss-of-function experiments to determine how BMSCs-derived EVs alleviate RA in vitro and in vivo. Methods: We isolated EVs from BMSCs and characterized them by transmission electron microscopy and western blot analysis. The regulatory relationship between miR-21 and TET1 was predicted by bioinformatics analysis and validated by dual luciferase assay. Next, we utilized bisulfite sequencing PCR to decipher how TET1 promoted KLF4 transcription. Then, we established an RA mouse model and determined the role of miR-21 in RA progression. Functional assays were used to validate the role the miR-21-TET1-KLF4 regulatory axis in controlling mouse fibroblast-like synoviocytes (mFLS) cell proliferation and inflammatory cytokines secretion in vitro. Results: RT-qPCR results revealed that miR-21 was highly expressed in BMSCs-derived EVs, and confirmed that BMSCs-derived EVs transferred miR-21 into mFLS cells. Bioinformatic analysis predicted that TET1 was the directly downstream target of miR-21, which was further validated by dual luciferase assay. TET1 promoted KLF4 promoter methylation to increase its expression. Collectively, BMSCs-derived EVs relieved RA by delivering miR-21, while the exosomal miR-21 alleviated RA through targeting the TET1/KLF4 regulatory axis. Conclusion: miR-21 from BMSCs-derived EVs suppresses KLF4 to relive RA by targeting TET1.

Bone Mesenchymal Stem Cell-Derived sEV-Encapsulated Thermosensitive Hydrogels Accelerate Osteogenesis and Angiogenesis by Release of Exosomal miR-21

“…Furthermore, the absorbed sEVs exhibited osteogenic and angiogenic properties. Previous studies have also reported that angiogenic activity was also promoted by miR-21 overexpression in BMSC-sEVs ( Wu et al, 2020 ; Li G.-Q. et al, 2021 ; Zhang et al, 2021 ).…”

Development of Extracellular Vesicle Therapeutics: Challenges, Considerations, and Opportunities

“…Native EVs are cell-derived, lipid-bound nanoparticles (30–1000 nm) that facilitate intercellular communication through targeted delivery of bioactive cargo to alter recipient cell phenotype ( Al-Nedawi et al, 2008 ; Skog et al, 2008 ; Alvarez-Erviti et al, 2011 ; Peinado et al, 2012 ; Kamerkar et al, 2017 ; Wu et al, 2019 ; Cocozza et al, 2020 ; Li et al, 2021b ) [reviewed in Vader et al (2016) , Murphy et al (2019) , and Russell et al (2019) ]. EVs are involved in transferring molecular cargo, between animal cells ( Valadi et al, 2007 ; Mittelbrunn et al, 2011 ; Ono et al, 2019 ), from parasite to its mammalian host ( Buck et al, 2014 ; Coakley et al, 2017 ), and from plant to fungal cells ( Cai et al, 2018 ).…”

“…Preclinical development of EVs have focused on their ability to horizontally transfer bioactive components, eliciting transcriptional and translational modulation ( Valadi et al, 2007 ), antigen presentation ( Gurnani et al, 2004 ; Garcia et al, 2016 ), and functional regulation in recipient cells ( Al-Nedawi et al, 2008 ; Skog et al, 2008 ; Alvarez-Erviti et al, 2011 ; Peinado et al, 2012 ; Kamerkar et al, 2017 ; Wu et al, 2019 ; Zhang Q. et al, 2019 ; Cocozza et al, 2020 ; Li et al, 2021b ). A landmark study demonstrated that embryonic stem cell derived EVs contribute to epigenetic reprogramming of target cells ( Ratajczak et al, 2006 ); EVs were enriched in key players for early pluripotency, hematopoietic stem cells (Scl, HoxB4 and GATA 2), and induced MAPK/Akt phosphorylation.…”

“…Lung-based pathologies are employing inhalation as the method of administration to ensure site-specific delivery (NCT04389385, NCT04747574, and NCT04276987). Injections of EVs to sites of damage are another method of focusing delivery, with therapeutic application of EVs directly to arthritic joints ( Wang et al, 2017 ; Wu et al, 2019 ; Li et al, 2021b ), venous ulcers (NCT04652531), inner ears ( Gyorgy et al, 2017 ), eyes (NCT03437759), and heart [damaged myocardium ( Yao J. et al, 2021 )]. While suitable for individual treatments at injection-accessible sites, ongoing release of EV-based therapeutics at inaccessible sites raises a challenge.…”

Extracellular Vehicles of Oxygen-Depleted Mesenchymal Stromal Cells: Route to Off-Shelf Cellular Therapeutics?

“…In addition, BMSC EVs seem to mediate similar anti-inflammatory properties than that of their parent cells [ 103 ]. These effects, however, are mediated, at least partly, by different miRNA populations including the miRNA-21 and miRNA-34a that target KLF4 and the cyclin I/ATM/ATR/p53 axis, respectively [ 104 , 105 ]. Besides their anti-inflammatory effects in chronic inflammatory conditions, BMSC-derived EVs also display similar activities in graft versus host models by influencing the ratio of T cell populations biasing regulatory over cytotoxic species [ 106 ].…”