Hypermethylation of microRNA‐149 activates SDF‐1/CXCR4 to promote osteogenic differentiation of mesenchymal stem cells

Abstract

MicroRNAs (miRs) involve in osteogenic differentiation and osteogenic potential of mesenchymal stem cells (MSCs). Accordingly, the present study aimed to further uncover role miR-149 plays in osteogenic differentiation of MSCs with the involvement of the stromal cell-derived factor-1 (SDF-1)/CXC chemokine receptor-4 (CXCR4) pathway.Initially, the osteogenic differentiation model was induced. Next, the positive expression of STRO-1 in periosteum, alkaline phosphatase (ALP) activity, osteocalcin (OCN) protein content, and the calcium deposition in MSCs were determined. MSCs were treated with DNA methyltransferase inhibitor 5-aza-CdR, SDF-1 neutralizing antibody, or CXCR4 antagonist AMD3100 to investigate their roles in osteogenic differentiation; with the expression of CD44, CD90, CD14, and CD45 detected. Furthermore, the levels of SDF-1 and CXCR4, and the genes related to stemness (Nanog, Oct-4, and Sox-2) were measured to explore the effects of miR-149. The obtained data revealed the upregulation of STRO-1 in the periosteum. miR-149 could specifically bind to SDF-1. Besides, increased miR-149 methylation, higher ALP activity and OCN content, decreased positive rates of CD44 and CD90, and increased positive rates of CD14 and CD45 were found in osteogenic differentiation of MSCs. Subsequently, 5-Aza-CdR treatment reversed the abovementioned effects. MSCs were finally treated with SDF-1 neutralizing antibody or AMD3100 to decrease Nanog, Oct-4, and Sox-2 expression. Taken together these results, miR-149 hypermethylation has the potential to activate the SDF-1/CXCR4 pathway and further promote osteogenic differentiation of MSCs.

SDF-1/CXCR4 Axis-mediated Migration of Systematically Transplanted Bone Marrow Mesenchymal Stem Cells Towards the Injured Spinal Cord in a Rat Model

“…Among these components, SDF-1 and its receptor CXCR4 are crucial for bone marrow retention, mobilization, and are involved in homing and recruitment of stem cells to sites of injured tissues [17]. SDF-1 has been reported to be expressed in a wide range of issues, including those of the heart, skeleton, live, pancreas, endothelium, skin, and spinal cord [20,[32][33][34][35][36]. Recent studies suggest that the interaction of SDF-1 and CXCR4 could partially mediate the tra cking of MSCs to the impaired nucleus in the brain [20,37].…”

DNA methylation of noncoding RNAs: new insights into osteogenesis and common bone diseases

“…Another article confirmed the effect of miR-342-3p in osteogenic differentiation and explored the regulation mechanism between miR-342-3p and DNA methylation of its hosting gene EVL [37]. They found hypomethylation in the promoter of EVL promoted the miR-342-3p expression in osteogenic differentiation, Li et al [35] miRNA miR-29b MiR-29b targeted DNMT1 and led to the methylation modification in the Notch1 promoter, which affected the Notch signaling pathway and regulated the osteogenic differentiation in BMSCs.…”

“…It revealed that miR-149 regulated MSCs osteogenic differentiation through epigenetic modifications ( Fig. 1) [35].…”

“…Curtis et al [40] ncRNAs noncoding RNAs, DNMT DNA methyltransferase enzymes, BMSCs bone marrow stromal cells, hMSCs human mesenchymal stem cells Fig. 1 Hypermethylation at the CpG sites of miR-149 promoter inhibited the expression of miR-149, thereby eliminating the inhibitory effect on SDF-1 and activating the SDF-1/CXCR4 signaling, which ultimately induced osteogenic differentiation of MSCs [35] while hypermethylation associated with low expression of miR-342-3p in the undifferentiated group.…”

Emerging trends in chromatin remodeler plasticity in mesenchymal stromal cell function

“…143 Further studies suggested that miR-149 mediated osteogenesis was steered by methyltransferases. 144 Contrasting findings regarding the role of Dnmts were reported during adipogenesis. While inhibition of Dnmt in multipotent C3H10T1/2 cells and 3T3-L1 preadipocytes favored adipogenesis, 145,146 upregulation of canonical Wnt signaling was demonstrated upon Dnmt inhibition in 3T3-L1 and ST2 mesenchymal precursor cells hindering adipogenesis.…”