Identification of candidate genes simultaneously shared by adipogenesis and osteoblastogenesis from human mesenchymal stem cells
Identification of candidate genes simultaneously shared by adipogenesis and osteoblastogenesis from human mesenchymal stem cells
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人间充质干细胞脂肪生成和成骨细胞生成同时共享的候选基因的鉴定
DOI：
10.5603/fhc.a2022.0012
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发表时间：
2022
期刊：
影响因子：
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通讯作者：
Xiaoyuan Xu
中科院分区：
文献类型：
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作者：
Xia Yi;Ping Wu;Yunyan Fan;Ying Gong;Jianyun Liu;Jianjun Xiong;Xiaoyuan Xu
在骨质疏松症领域，临床已经很好地确立了骨形成与脂质积累之间的一定关系。虽然已经有许多分子被证明与脂肪形成和成骨细胞形成有关，但相互的转录调控仍有待探索。在这里，我们试图通过RNA-Seq技术，利用人间充质干细胞（hMSCs）在3、5和7天鉴定脂肪细胞/成骨细胞分化的共同候选基因。采用RNA干扰（RNAi）技术，通过油红/茜素红/碱性磷酸酶（ALPL）染色和qRT-PCR（实时荧光定量PCR）进一步证实候选基因在脂肪成骨过程中的功能。鉴定的275个显著差异表达基因（DEGs），特别是下调基因最为普遍，PI3K-AKT信号通路最为丰富，同时被两个分化事件共享。利用慢病毒系统，我们进一步证实ANKRD1（锚蛋白重复结构域1）通过RNA干扰（RNAi）促进脂肪形成并抑制成骨细胞形成，而IGF1（胰岛素样生长因子1）基于生物标志物的基因表达和蛋白积累以及细胞表型特性同时促进脂肪成骨细胞形成。本研究将为控制脂肪/成骨细胞平衡或hMSCs命运选择提供潜在的分子开关，并为代谢性骨病骨质疏松症的研究和治疗靶点筛选提供线索。
In osteoporosis field, it had been clinically well established a given relationship between bone formation and lipid accumulation. Although numerous molecules had been well documented for adipogenesis and osteoblastogenesis (adipo-osteoblastogenesis), the reciprocal transcriptional regulation still remained to be explored.Here, we tried to identify the common candidate genes of adipocyte/osteoblastocyte differentiation at 3, 5, and 7 days using human mesenchymal stem cells (hMSCs) via RNA-Seq technique. By using RNA interference (RNAi), we further confirmed the function of candidate genes during adipo-osteoblastogenesis through Oil Red/Alizarin Red/alkaline phosphatase (ALPL) staining and qRT-PCR (quantitative real-time PCR).The identified 275 significantly differentially expressed genes (DEGs), especially with the down-regulated genes most prevalent and PI3K-AKT signaling pathway mostly enriched, were simultaneously shared by both differentiation events. Using lentiviral system, we further confirmed that ANKRD1 (ankyrin repeat domain 1) promoted adipogenesis and inhibited osteoblastogenesis via RNA interference (RNAi), and IGF1 (insulin like growth factor 1) simultaneously facilitated adipo-osteoblastogenesis on the base of gene expression and protein accumulation of biomarkers and cellular phenotype property.This study would provide the potential molecular switches to control the adipocyte/osteoblastocyte balance or hMSCs fate choices and clues to screen the study and therapy targets of metabolic bone disease osteoporosis.