MFN2 knockdown promotes osteogenic differentiation of iPSC-MSCs through aerobic glycolysis mediated by the Wnt/β-catenin signaling pathway

Abstract

Background Mitofusin-2 (MFN2) is a kind of GTPase that participates in the regulation of mitochondrial fusion, which is related to a variety of physiological and pathological processes, including energy metabolism, cell differentiation, and embryonic development. However, it remains unclear whether MFN2 is involved in the metabolism and osteogenic differentiation of mesenchymal stem cells (MSCs). Methods MFN2 knockdown (MFN2-KD) and MFN2-overexpressing (MFN2-OE) induced pluripotent stem cell-derived mesenchymal stem cells (iPSC-MSCs) were constructed by lentivirus. The commercial kits were utilized to detect the glycolysis and oxidative phosphorylation (OXPHOS) rate. Flow cytometry, Western blot, quantitative real-time polymerase chain reaction (qRT-PCR), RNA-seq, immunofluorescence, and immunoprecipitation were employed for phenotype and molecular mechanism assessment. Results We demonstrated that MFN2 and Wnt/β-catenin signaling pathway regulated glycolysis of iPSC-MSCs. The lack of MFN2 promoted the osteogenic differentiation of iPSC-MSCs, and aerobic glycolysis in the presence of sufficient oxygen, which increased glucose consumption and lactic acid production, as well as the glycolytic enzyme activity and gene expression. Inhibiting the Wnt/β-catenin signaling pathway normalized the enhanced glycolytic rate and osteogenic differentiation of MFN2-KD iPSC-MSCs. MFN2-OE iPSC-MSCs displayed the opposite phenotype. Conclusions Downregulating MFN2 promotes osteogenic differentiation of iPSC-MSCs through aerobic glycolysis mediated by the Wnt/β-catenin signaling pathway. Our research reveals the new function of MFN2 in regulating the osteogenic differentiation and energy metabolism of MSCs, which will provide a new therapeutic target and theoretical basis for alveolar bone repair and periodontal regenerative treatment.

Eucommia granules activate Wnt/β-catenin pathway, and improve oxidative stress, inflammation, and endothelial injury in preeclampsia rats

“…The canonical Wnt/β-catenin pathway is triggered when the Wnts are combined with Wnt ligands (e.g., Wnt1). The expression of Wnt1 is higher in trophoblast cells of early pregnancy than in trophoblast cells of late pregnancy, indicating that Wnt1 can regulate trophoblast invasion 31 . β-catenin is a key factor in the Wnt/β-catenin pathway.…”

Icariin ameliorates glycolytic dysfunction in Alzheimer's disease models by activating the Wnt/β‐catenin signaling pathway

“…Recent data manifest that the glycolysis in iPSC-MSCs is regulated by canonical Wnt signaling pathway [64] and Wnt activators effectively improved learning cognitive function in APP/PS1 double transgenic AD mice by increasing the activity of the glycolytic key enzyme via upregulating the canonical Wnt signaling pathway [14]. In this study, when the canonical Wnt signaling pathway was inhibited by DKK1, we manifest that the effect of ICA on the glycolysis in HT22 cells damaged by Ab 25-35 was abolished (Fig.…”

Downregulation of Mitochondrial Fusion Protein Expression Affords Protection from Canonical Necroptosis in H9c2 Cardiomyoblasts

“…Since the knockdown of Mfn1/2 and Opa1 had no effect on TAK1 mRNA levels, post-transcriptional modifications and protein-protein interactions may play a role. Fourth, considering the partial block of protection afforded by Mfn1/2 and Opa1 knockdown mediated by TAK1 inhibition, further detailed investigations are needed to demonstrate the underlying mechanism: Mfn2 ablation-induced upregulation of the glycolytic pathway and mTORC2-Akt signaling are possible candidates [ 40 , 41 , 42 , 43 ]. Fifth, there are conflicting results about the significance of TAK1 in the cardiomyocytes subjected to pathological stress: TAK1 activation is a promoter of pressure overload-induced cardiac dysfunction, whereas TAK1 ablation is detrimental for a pressure-overloaded heart through the enhancement of necroptotic signaling [ 37 , 44 , 45 ].…”