Overexpression of MiR-335-5p Promotes Bone Formation and Regeneration in Mice.
Overexpression of MiR-335-5p Promotes Bone Formation and Regeneration in Mice.
复制标题
DOI：
10.1002/jbmr.3230
复制
发表时间：
2017-12
期刊：
影响因子：
--
通讯作者：
Chen J
中科院分区：
文献类型：
--
作者：
Zhang L;Tang Y;Zhu X;Tu T;Sui L;Han Q;Yu L;Meng S;Zheng L;Valverde P;Tang J;Murray D;Zhou X;Drissi H;Dard MM;Tu Q;Chen J
microRNAs（miRNAs）和Wnt信号通路在调节骨发育和体内平衡中起着关键作用。我们先前的研究揭示了miR-335- 5 p在小鼠胚胎成骨细胞和肥大软骨细胞中的高表达，以及miR-335- 5 p通过下调Wnt拮抗剂Dickkopf-1（DKK 1）促进成骨分化的能力。本研究的目的是研究miR-335- 5 p组成型过表达对体内骨形成和再生的影响。为此，我们通过osterix启动子产生了在成骨细胞谱系中特异性过表达miR-335- 5 p的转基因小鼠系，并表征了其骨表型。骨组织形态计量学和μCT分析显示，与野生型同窝小鼠相比，转基因小鼠的骨量更高，骨形成参数增加。转基因小鼠骨中增加的骨量也与成骨分化标志物的表达增强相关。成骨诱导后，从转基因小鼠中分离的BMSCs显示出比野生型小鼠BMSCs培养物更高的成骨标志物mRNA表达。在成骨诱导后转基因小鼠的BMSC培养物中Runx 2和Osx的蛋白表达也上调，而DKK 1的蛋白表达下调。最重要的是，来自转基因小鼠的BMSCs能够修复颅面骨缺损，如通过用BMSCs处理的缺损中新形成的骨的μCT分析、H&E染色和OCN免疫组织化学所证明的。综上所述，我们的研究结果表明，在成骨细胞谱系中由osterix启动子驱动的miR-335- 5 p的组成型过表达诱导小鼠的成骨分化和骨形成，并支持miR-335- 5 p修饰的BMSC在颅面骨再生中的潜在应用。
MicroRNAs (miRNAs) and the Wnt signaling pathway play critical roles in regulating bone development and homeostasis. Our previous study revealed high expression of miR-335-5p in osteoblasts and hypertrophic chondrocytes in mouse embryos and the ability of miR-335-5p to promote osteogenic differentiation by downregulating Wnt antagonist Dickkopf-1 (DKK1). The purpose of this study was to investigate the effects of miR-335-5p constitutive overexpression on bone formation and regeneration in vivo. To that end, we generated a transgenic mouse line specifically over-expressing miR-335-5p in osteoblasts lineage by the osterix promoter and characterized its bone phenotype. Bone histomorphometry and μCT analysis revealed higher bone mass and increased parameters of bone formation in transgenic mice than in wild-type littermates. Increased bone mass in transgenic mice bones also correlated with enhanced expression of osteogenic differentiation markers. Upon osteogenic induction, BMSCs isolated from transgenic mice displayed higher mRNA expression of osteogenic markers than wild-type mice BMSCs cultures. Protein expression of Runx2 and Osx was also upregulated in BMSC cultures of transgenic mice upon osteogenic induction, while that of DKK1 was down-regulated. Most importantly, BMSCs from transgenic mice were able to repair craniofacial bone defects as demonstrated by μCT analysis, H&E staining and OCN immunohistochemistry of newly formed bone in defects treated with BMSCs. Taken together, our results demonstrate constitutive overexpression of miR-335-5p driven by an osterix promoter in the osteoblast lineage induces osteogenic differentiation and bone formation in mice and support the potential application of miR-335-5p-modified BMSCs in craniofacial bone regeneration.
登录
查看更多内容
影响因子：
64.8
作者：
Glinka, A;Wu, W;Niehrs, C
通讯作者：
Niehrs, C
影响因子：
6.2
作者：
van der Horst, G;van der Wert, SM;Karperien, M
通讯作者：
Karperien, M
影响因子：
64.5
作者：
Gong, YQ;Slee, RB;Warman, ML
通讯作者：
Warman, ML
影响因子：
6.2
作者：
Valverde, Paloma;Zhang, Jin;Chen, Jake
通讯作者：
Chen, Jake
影响因子：
4.6
作者：
MacDonald, BT;Adamska, M;Meisler, MH
通讯作者：
Meisler, MH