Improved calvarial bone repair by hASCs engineered with Cre/loxP-based baculovirus conferring prolonged BMP-2 and MiR-148b co-expression
Improved calvarial bone repair by hASCs engineered with Cre/loxP-based baculovirus conferring prolonged BMP-2 and MiR-148b co-expression
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DOI：
10.1002/term.2208
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发表时间：
2017-11-01
影响因子：
3.3
通讯作者：
Hu, Yu-Chen
中科院分区：
文献类型：
--
作者：
Li, Kuei-Chang;Lo, Shih-Chun;Hu, Yu-Chen
修复大的颅骨缺损仍然是一项具有挑战性的任务。以前，发现miR-148 b与骨形态发生蛋白2（BMP-2）协同作用时，增强了人脂肪源性干细胞（hASC）的成骨作用，并改善了裸鼠颅骨愈合。然而，miR-148 b的分子靶点仍然难以捉摸。本文揭示了miR-148 b直接靶向NOG，其基因产物（noggin）是BMP的拮抗剂并负调节BMP诱导的成骨分化和骨形成。采用新的基于Cre/loxP的杆状病毒系统来驱动hASC中BMP-2和miR-148 b的延长过表达，其中BMP-2过表达诱导头蛋白表达，但同时miR-148 b表达下调头蛋白，从而缓解负调控环并改善hASC成骨，而不阻碍hASC增殖或触发明显的细胞毒性。将共表达BMP-2和miR-148 b的工程化hASC植入裸鼠体内，能够在移植后12周基本修复临界尺寸的颅骨骨缺损（直径4 mm），填充83%的缺损面积，75%的骨体积，并将骨密度恢复至原始骨密度的89%。这种上级愈合效果表明基于Cre/loxP的杆状病毒介导的BMP-2/miR-148 b表达用于颅骨骨修复的潜力。版权所有（c）2016约翰威利父子有限公司
Repairing large calvarial bone defects remains a challenging task. Previously, it was discovered that that miR-148b, when acting in concert with bone morphogenetic protein 2 (BMP-2), enhanced the osteogenesis of human adipose-derived stem cells (hASCs) and improved calvarial bone healing in nude mice. However, the molecular target of miR-148b remained elusive. Here it is revealed that miR-148b directly targets NOG, whose gene product (noggin) is an antagonist to BMPs and negatively regulates BMP-induced osteogenic differentiation and bone formation. A new Cre/loxP-based baculovirus system was employed to drive prolonged BMP-2 and miR-148b overexpression in hASCs, wherein the BMP-2 overexpression induced noggin expression but the concurrent miR-148b expression downregulated noggin, thus relieving the negative regulatory loop and ameliorating hASC osteogenesis without hindering hASC proliferation or triggering appreciable cytotoxicity. Implantation of the engineered hASCs coexpressing BMP-2 and miR-148b into nude mice enabled substantial repair of critical-size calvarial bone defects (4 mm diameter) at 12 weeks post-transplantation, filling 83% of the defect area, 75% of bone volume and restoring the bone density to 89% of the original bone density. Such superior healing effects indicate the potential of the Cre/loxP-based baculovirus-mediated BMP-2/miR-148b expression for calvarial bone repair. Copyright (c) 2016 John Wiley & Sons, Ltd.