Optogenetic control of mesenchymal cell fate towards precise bone regeneration
Optogenetic control of mesenchymal cell fate towards precise bone regeneration
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光遗传学控制间充质细胞命运以实现精确的骨再生
DOI：
10.7150/thno.36455
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发表时间：
2019-01-01
期刊：
影响因子：
12.4
通讯作者：
Zhou, Chen
中科院分区：
文献类型：
--
作者：
Wang, Weicai;Huang, Delan;Zhou, Chen
原理：细胞命运的时空控制在体内是非常重要的再生医学。目前，仍然没有实际的策略来调整细胞命运的时空。光遗传学是一种广泛应用于通过光以时空特异性的方式控制遗传定义的神经元中的细胞活动的生物学技术。方法：将骨髓间充质干细胞增殖和分化的主控基因Lhx8和BMP 2基因重组到定制的光遗传学调控系统中，构建成骨组织再生的调控系统。结果：与预期的一样，在光遗传控制系统感染的细胞中，蓝光诱导BMP 2表达，而Lhx8表达失活。BMP 2和Lhx8表达的光遗传学控制在体外反向调节MSC命运。通过动物实验，我们发现蓝光可以在体内微调再生。蓝光照射可显著促进骨髓间充质干细胞增殖和分化，并能显著促进骨髓间充质干细胞增殖和分化。结论：光遗传学调控骨髓间充质干细胞增殖和分化的主基因可能是一种精确再生医学的候选策略。
Rationale: Spatial-temporal control of cell fate in vivo is of great importance for regenerative medicine. Currently, there remain no practical strategies to tune cell-fate spatial-temporally. Optogenetics is a biological technique that widely used to control cell activity in genetically defined neurons in a spatiotemporal-specific manner by light. In this study, optogenetics was repurposed for precise bone tissue regeneration.Methods: Lhx8 and BMP2 genes, which are considered as the master genes for mesenchymal stem cell proliferation and differentiation respectively, were recombined into a customized optogenetic control system. In the system, Lhx8 was constitutively expressed, while BMP2 together with shLhx8 expression was driven by blue light.Results: As expected, blue light induced BMP2 expression and inactivated Lhx8 expression in cells infected with the optogenetic control system. Optogenetic control of BMP2 and Lhx8 expression inversely regulates MSC fate in vitro. By animal study, we found that blue light could fine-tune the regeneration in vivo. Blue light illumination significantly promotes bone regeneration when the scaffold was loaded with MSCs infected with adeno-Lhx8, GI-Gal4DBD, LOV-VP16, and BMP2-shLhx8.Conclusions: Together, our study revealed that optogenetic control of the master genes for mesenchymal stem cell proliferation and differentiation would be such a candidate strategy for precise regenerative medicine.