miR‐101‐loaded exosomes secreted by bone marrow mesenchymal stem cells requires the FBXW7/HIF1α/FOXP3 axis, facilitating osteogenic differentiation

Abstract

Exosomes derived from mesenchymal stem cells (MSCs) have emerged as significant mediators of intercellular communication, with studies highlighting their role in the transmission of biological signals between cells. Dominant microRNA (miRNA)-mediated translational repression of messenger RNAs has been extensively investigated in regard to its influence in orchestrating osteogenic differentiation. In the current study, we sought to ascertain the contributory role of miRNA-101 (miR-101) encapsulated in the process of bone marrow mesenchymal stem cell (BMSC)-derived exosomes in osteogenic differentiation. Exosomes were initially extracted from BMSCs at Days 0, 3, 12, and 21 of osteogenic differentiation by ultracentrifugation. Artificial modulation of miR-101 and FBXW7 (silencing and overexpression) were performed in the BMSCs to identify its effects on osteogenic factors, alkaline phosphatase activity, and osteogenic differentiation. Mechanistic exploration was performed to evaluate the binding affinity between miR-101 and FBXW7, the FBXW7-mediated HIF1α ubiquitination, and the HIF1α enrichment in the FOXP3 promoter region. Exosomes from MSCs in the late stage of osteogenic differentiation exhibited enhanced osteogenic differentiation. Upregulated miR-101 in MSC-derived exosomes was detected during osteogenic differentiation, while diminished levels of FBXW7 expression was noted. Importantly, miR-101 was found to specifically bind to the 3′-untranslated region of FBXW7. Meanwhile, data was obtained indicating that FBXW7 could ubiquitinate and degrade HIF1α to repress its upregulation during osteogenic differentiation. HIF1α bound to the promoter region of FOXP3 to facilitate osteogenic differentiation. Ultimately, the findings of the current study demonstrate that BMSC-derived exosomal miR-101 augments osteogenic differentiation in MSCs by inhibiting FBXW7 to regulate the HIF1α/FOXP3 axis.

miR-330–5p in Small Extracellular Vesicles Derived From Plastrum testudinis-Preconditioned Bone Mesenchymal Stem Cells Attenuates Osteogenesis by Modulating Wnt/β-Catenin Signaling

“…Nevertheless, serum levels of bone turnover markers could reflect the status of bone metabolism to some degree, but there was no significant difference between the OVX and PT-sEV groups. Higher fat content and increased BMI are other characteristics of OVX rats ( Nam et al, 2017 ; Li et al, 2021 ). In this study, PT-sEV diminished these phenotypes.…”

Educating EVs to Improve Bone Regeneration: Getting Closer to the Clinic

“…The second specific condition determining the osteoanabolic potential of EVs is the osteogenic differentiation stage of BM-MSCs [56]. Strikingly, recent works have pointed out that EVs derived from MSCs in the late state of osteogenic differentiation (day 21) have increased osteoanabolic potential when compared to EVs from undifferentiated MSCs or MSCs undergoing early osteogenic differentiation (day 3) [57]. The different cargo composition of EVs depending on the differentiation stage of MSCs was a possible explanation for this finding.…”

“…The authors demonstrated that EVs could be enriched in these miRNAs when overexpressed in parent MSCs, without affecting distinctive features of EVs such as morphology, size, and the expression of surface proteins CD9 and CD63, which are used as EV markers. Moreover, these EVs improved the osteogenic differentiation of MSCs and enhanced bone regeneration in animal models of bone defects [57,91]. The overexpression of pro-osteogenic proteins in parent MSCs has also been considered as a strategy to enhance the osteoanabolic potential of EVs.…”

Noncoding RNA in Extracellular Vesicles Regulate Differentiation of Mesenchymal Stem Cells

“…miR-21/Smad7 axis detected by the exosome of BMSC is the mechanism of negative osteogenesis in osteoporosis patients (100). After 21 days of osteogenic inducement, miR-101 is upregulated in BMSC-derived exosomes, and the target demonstrates osteogenic potential upon functional verification (101). miR-335/VapB/Wnt/β-catenin axis promotes bone fracture recovery and osteoblast differentiation, which is attributed to BMSC-derived EVs (102).…”