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

• Published in: Stem cell research & therapy Vol. 13; no. 1; p. 162
• Main Authors: Deng, Lidi, Yi, Siqi, Yin, Xiaohui, Li, Yang, Luan, Qingxian
• Format: Journal Article
• Language: English
• Published: England BioMed Central 12-04-2022; BMC
• Subjects: Ablation; Acid production; Aerobic glycolysis; Alveolar bone; Antibodies; beta Catenin - metabolism; Bone healing; Cell culture; Cell cycle; Cell differentiation; Cell Differentiation - genetics; Cells, Cultured; Dehydrogenases; DNA-directed RNA polymerase; Embryogenesis; Energy metabolism; Enzymatic activity; Enzymes; Ethanol; Female; Fibroblasts; Flow cytometry; Gene expression; Genes; Glucose; Glycolysis; Glycolysis - genetics; GTP Phosphohydrolases - genetics; GTP Phosphohydrolases - metabolism; Humans; Immunofluorescence; Immunoprecipitation; Induced pluripotent stem cells; Induced Pluripotent Stem Cells - metabolism; Infections; Kinases; Lactic acid; Mesenchymal stem cells; Mesenchymal Stem Cells - metabolism; Mesenchyme; Metabolism; Metabolites; Mitochondria; Mitochondrial Proteins - genetics; Mitochondrial Proteins - metabolism; Mitofusin-2; Osteogenesis; Osteogenic differentiation; Oxidative phosphorylation; Oxygen consumption; Phenotypes; Phosphorylation; Polymerase chain reaction; Pregnancy; Reagents; Signal transduction; Stem cells; Wnt protein; Wnt Signaling Pathway - genetics; Wnt/β-catenin signaling pathway; β-Catenin; Canada; United States > US; China; Japan; Mitofusin-2; Aerobic glycolysis; Wnt/β-catenin signaling pathway; Induced pluripotent stem cells; Osteogenic differentiation; Mesenchymal stem cells
• ISSN: 1757-6512; 1757-6512
• DOI: 10.1186/s13287-022-02836-w

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).
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.
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.
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.