Regulating Osteogenic Differentiation by Suppression of Exosomal MicroRNAs
Regulating Osteogenic Differentiation by Suppression of Exosomal MicroRNAs
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DOI：
10.1089/ten.tea.2018.0257
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发表时间：
2019-08-01
影响因子：
4.1
通讯作者：
Lee, Heon-Jin
中科院分区：
文献类型：
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作者：
Choi, Song-Yi;Han, Eun-Chong;Lee, Heon-Jin
外泌体是天然运输生物分子（包括miRNA）的膜状纳米囊泡，并且在细胞间通信中发挥关键作用。我们发现前成骨细胞外泌体的生长条件环境含有高水平的let-7 miRNA。在阻断外泌体分泌后，我们发现前成骨细胞（MC 3 T3-E1细胞）的成骨分化显著降低，而从生长条件中添加外泌体恢复成骨能力。通过let-7模拟物或抑制剂的异位表达改变MC 3 T3-E1细胞中成骨标志物的表达水平。这些发现表明，这是由于外泌体中的let-7而发生的。此外，let-7通过电穿孔被抑制剂灭活的外泌体有效地失去了它们的成骨分化能力。因此，遗传修饰的外泌体可以作为用于发育控制（包括骨生成调节）的高效生物材料。我们研究了外泌体在骨生成中的作用以及使用miRNA转染的外泌体来控制成骨分化。外泌体miRNAs的RNA测序（RNA-seq）显示，前成骨细胞外泌体环境的生长条件具有高水平的let-7，其在成骨调控中起关键作用。我们通过在MC 3 T3-E1细胞中在生长条件下将let-7抑制剂引入exosomes中来修饰exosomes，并揭示了通过工程改造使let-7失活的exosomes失去恢复成骨分化的能力。基因修饰的外泌体可以作为强大的生物材料，用于发育控制，包括骨生成调节。
Exosomes are membranous nanovesicles that naturally transport biomolecules, including miRNAs, and play critical roles in intercellular communication. We found that the growth condition milieu of preosteoblast exosomes contains a high level of the let-7 miRNAs. After blocking exosome secretion, we found that osteogenic differentiation of preosteoblasts (MC3T3-E1 cells) decreased significantly whereas addition of exosomes from growth condition recovered osteogenic ability. The expression levels of osteogenic markers in MC3T3-E1 cells were changed through the ectopic expression of the let-7 mimics or inhibitors. These findings suggest that this occurred due to the let-7 in the exosomes. Furthermore, exosomes whose let-7 was inactivated by inhibitors via electroporation efficiently lost their osteogenic differentiation capacity. Genetically modified exosomes may thus serve as highly effective biomaterials for developmental control, including osteogenesis regulation. Impact Statement We investigated the role of exosomes in osteogenesis and the use of miRNA inhibitor-transfected exosomes to control osteogenic differentiation. RNA-sequencing (RNA-seq) of exosomal miRNAs revealed that growth condition of milieu of preosteoblast exosomes harbors high levels of let-7, which plays a critical role in osteogenesis regulation. We modified exosomes by transfecting let-7 inhibitor into exosomes under growth condition in MC3T3-E1 cells and revealed that exosomes whose let-7 was inactivated by engineering lost the ability to recover osteogenic differentiation. Genetically modified exosomes may serve as powerful biomaterials for developmental control, including of osteogenesis regulation.