Extracellular vesicle-encapsulated miR-22-3p from bone marrow mesenchymal stem cell promotes osteogenic differentiation via FTO inhibition

Abstract

Background: Bone marrow mesenchymal stem cells (BMSCs) exhibit the capacity to self-renew and differentiate into multi-lineage cell types, including osteoblasts, which are crucial regulators of fracture healing. Thus, this study aims to investigate the effect of microRNA (miR)-22-3p from BMSC-derived EVs on osteogenic differentiation and its underlying mechanism. Methods: Extracellular vesicles (EVs) were isolated from BMSCs and taken up with BMSCs. Dual-luciferase reporter gene assay was used to verify the binding relationship between miR-22-3p and FTO. Loss-and gain-of-function experiments were performed to determine the roles of EV-delivered miR-22-3p and FTO in osteogenic differentiation as well as their regulatory role in the MYC/PI3K/AKT axis. To determine the osteogenic differentiation, ALP and ARS stainings were conducted, and the levels of RUNX2, OCN, and OPN level were determined. In vivo experiment was conducted to determine the function of EV-delivered miR-22-3p and FTO in osteogenic differentiation, followed by ALP and ARS staining. Results: miR-22-3p expression was repressed, while FTO expression was elevated in the ovariectomized mouse model. Overexpression of miR-22-3p, EV-delivered miR-22-3p, increased ALP activity and matrix mineralization of BMSCs and promoted RUNX2, OCN, and OPN expressions in BMSCs. miR-22-3p negatively targeted FTO expression. FTO silencing rescued the suppressed osteogenic differentiation by EV-delivered miR-22-3p inhibitor. FTO repression inactivated the MYC/PI3K/AKT pathway, thereby enhancing osteogenic differentiation both in vivo and in vitro. Conclusion: In summary, miR-22-3p delivered by BMSC-derived EVs could result in the inhibition of the MYC/PI3K/ AKT pathway, thereby promoting osteogenic differentiation via FTO repression.

Molecular consequences of fetal alcohol exposure on amniotic exosomal miRNAs with functional implications for stem cell potency and differentiation

“…This indicates that alcohol affects osteogenic differentiation through altering exosomes, making miRNAs a very likely candidate for such effect. Previous studies have reported an association between elevated miRNA 22-3p levels and increased expression of RUNX2 and OCN [67]. Therefore, a decrease in miRNA 22-3p can lead to a decrease in these osteogenic markers, which was also observed in our experiments (Fig 5D).…”

“…Therefore, up-regulation of these miRNAs observed in our results ( Figs 3D and 4A) can inhibit osteogenic differentiation. In contrast, EV-encapsulated miRNA-22-3p from BMSCs have been shown to promote osteogenic differentiation in neighboring BMSCs and increase matrix mineralization [67]. Hence, the upregulation of miRNAs 25-3p and 214-3p as well as down-regulation of miRNA 22-3p all can lead to inhibition of osteogenic gene expression observed in Fig 5, prevent osteogenic differentiation, and cause the osteogenic deficits observed in FAE.…”

Regulatory Role of the RNA N6-Methyladenosine Modification in Immunoregulatory Cells and Immune-Related Bone Homeostasis Associated With Rheumatoid Arthritis

“…Osteoblast differentiation is also dependent on the regulation of FTO. Studies have shown that miR-22-3p and miR-149-3p modulate the osteoblast differentiation of BMSCs by targeting FTO (Li et al, 2019 ; Zhang et al, 2020 ). Mice with a global loss of FTO or a selective lack FTO in osteoblasts exhibit bone loss (Sachse et al, 2018 ; Zhang Q. et al, 2019 ).…”

Obesity regulates miR‐467/HoxA10 axis on osteogenic differentiation and fracture healing by BMSC‐derived exosome LncRNA H19

“…These regulatory effects of lncRNAs have also begun to attract widespread attention 11 . Recent studies have shown that some lncRNAs are involved in regulating the adipose differentiation and osteogenic differentiation of BMSCs 12,13 . Studies have reported that LncRNA H19 can stimulate the osteogenic differentiation of BMSCs, while inhibiting the adipogenic differentiation of BMSCs 14,15 .…”

“…11 Recent studies have shown that some lncRNAs are involved in regulating the adipose differentiation and osteogenic differentiation of BMSCs. 12,13 Studies have reported that LncRNA H19 can stimulate the osteogenic differentiation of BMSCs, while inhibiting the adipogenic differentiation of BMSCs. 14,15 Our previous research results indicate that LncRNA H19 adsorbs miR-188 through the ceRNA mechanism and then mediates LCoR's influence on the osteogenic and adipogenic differentiation of mouse BMSCs, 15 but does LncRNA H19 pass externally?…”