Vestigial-Like 3 Plays an Important Role in Osteoblast Differentiation by Regulating the Expression of Osteogenic Transcription Factors and BMP Signaling
Vestigial-Like 3 Plays an Important Role in Osteoblast Differentiation by Regulating the Expression of Osteogenic Transcription Factors and BMP Signaling
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DOI：
10.1007/s00223-022-00997-7
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发表时间：
2022-06-24
影响因子：
4.2
通讯作者：
Zhang，Bin
中科院分区：
文献类型：
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作者：
Yuan，Haoze;Ikegame，Mika;Zhang，Bin
我们以前的基因分析表明，转录辅因子残留样3（VGLL 3）基因表达上调的机械张力在小鼠颅缝，符合加速成骨细胞分化。因此，我们假设VGLL 3在成骨分化中起重要作用。为了阐明VGLL 3在成骨细胞中的功能，我们检测了其在小鼠骨组织和成骨细胞系MC 3 T3-E1中的表达特征。我们进一步研究了Vgll 3敲低对成骨细胞分化和骨形态发生蛋白（BMP）信号传导的影响。在小鼠颅缝中，膜骨化发生，VGLL 3主要在成骨细胞，前成骨细胞和成纤维细胞的细胞核中被免疫化学检测到。VGLL 3在MC 3 T3-E1细胞中的表达是短暂的，并且在分化的相对早期阶段达到峰值。RNA测序显示Vgl 13敲低细胞中下调的基因在成骨细胞分化相关的基因本体论术语中富集。有趣的是，大多数上调的基因与细胞分裂有关。靶向Vgll 3敲低显著抑制主要成骨转录因子（Runx 2、Sp 7/osterix和Dlx 5）的表达和成骨细胞分化。它还减弱BMP信号传导;此外，外源性BMP 2部分恢复Vgl 13敲低细胞中成骨转录因子的表达。此外，Vgll 3的过表达增加了成骨转录因子的表达。这些结果表明，VGLL 3在促进成骨细胞分化中起关键作用，并且该过程的一部分由BMP信号传导介导。进一步阐明VGLL 3的功能将增加我们对骨生成和骨骼疾病病因学的理解。
Our previous gene profiling analysis showed that the transcription cofactor vestigial-like 3 (VGLL3) gene expression was upregulated by mechanical tension in the mouse cranial suture, coinciding with accelerated osteoblast differentiation. Therefore, we hypothesized that VGLL3 plays a significant role in osteogenic differentiation. To clarify the function of VGLL3 in osteoblasts, we examined its expression characteristics in mouse bone tissue and the osteoblastic cell line MC3T3-E1. We further examined the effects ofVgll3knockdown on osteoblast differentiation and bone morphogenetic protein (BMP) signaling. In the mouse cranial suture, where membranous ossification occurs, VGLL3 was immunohistochemically detected mostly in the nucleus of osteoblasts, preosteoblasts, and fibroblastic cells. VGLL3 expression in MC3T3-E1 cells was transient and peaked at a relatively early stage of differentiation. RNA sequencing revealed that downregulated genes inVgll3-knockdown cells were enriched in gene ontology terms associated with osteoblast differentiation. Interestingly, most of the upregulated genes were related to cell division. TargetedVgll3knockdown markedly suppressed the expression of major osteogenic transcription factors (Runx2, Sp7/osterix, and Dlx5) and osteoblast differentiation. It also attenuated BMP signaling; moreover, exogenous BMP2 partially restore osteogenic transcription factors’ expression inVgll3-knockdown cells. Furthermore, overexpression ofVgll3increased the expression of osteogenic transcription factors. These results suggest that VGLL3 plays a critical role in promoting osteoblast differentiation and that part of the process is mediated by BMP signaling. Further elucidation of VGLL3 function will increase our understanding of osteogenesis and skeletal disease etiology.