Mitofusin-2(MFN2)是一种参与线粒体融合调控的GTP酶,这与各种生理和病理过程有关,包括能量代谢,细胞分化,和胚胎发育。然而,目前尚不清楚MFN2是否参与间充质干细胞(MSCs)的代谢和成骨分化。
通过慢病毒构建MFN2敲低(MFN2-KD)和MFN2过表达(MFN2-OE)诱导的多能干细胞来源的间充质干细胞(iPSC-MSCs)。商业试剂盒用于检测糖酵解和氧化磷酸化(OXPHOS)速率。流式细胞术,蛋白质印迹,定量实时聚合酶链反应(qRT-PCR),RNA-seq,免疫荧光,和免疫沉淀用于表型和分子机制评估。
我们证明了MFN2和Wnt/β-catenin信号通路调节iPSC-MSCs的糖酵解。MFN2的缺失促进了iPSC-MSCs的成骨分化,在有足够氧气的情况下进行有氧糖酵解,这增加了葡萄糖的消耗和乳酸的产生,以及糖酵解酶活性和基因表达。抑制Wnt/β-catenin信号通路可使MFN2-KDiPSC-MSCs的糖酵解速率和成骨分化增强正常化。MFN2-OEiPSC-MSC表现出相反的表型。
下调MFN2通过Wnt/β-catenin信号通路介导的有氧糖酵解促进iPSC-MSCs的成骨分化。我们的研究揭示了MFN2在调节MSCs成骨分化和能量代谢方面的新功能,为牙槽骨修复和牙周再生治疗提供新的治疗靶点和理论依据。
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.