AntimiR-34a增强脂肪干细胞对骨形成蛋白高效诱导成骨作用反应性的分子机制研究

The bone tissue engineering technology with adipose derived stem cells (hASC) and bone morphogenetic protein (BMP) is one of the research hot element in the treatment of facial bone defects. Our previous studies indicated that: as the donors’ epigenetic modifications vary greatly, hASCs have a lower response and uncertainty for BMP induction. Changing the hASCs’ differentiation trend into fat, and resetting their osteogenic differentiation pathway is the key to improve hASCs’ osteogenic induction reactivity with BMP. Our previous study found that antimiR-34a through this epigenetic modification can significantly enhance the expression of osteogenic related proteins when hASCs treated with BMP, so the antimiR-34a has a good application prospect. However, the molecular mechanism of antimiR-34a promoting hASC osteogenesis has not been reported. We hypothesized that antimiR-34a upregulates Notch-TAZ, NF-kappaB-TAZ pathway is a major mechanism for antimiR-34a to achieve its osteogenesis. In this study, we proposed by RNA sequencing, CHIP-sequencing, CoIP, RNA silencing, and inhibitors, etc. to elucidate the signal transduction mechanism of antimiR-34a promoting hASC osteogenesis, provide a theoretical basis for the effective construction of tissue engineered bone with hASCs and BMP.

以脂肪干细胞（hASCs）和骨形成蛋白（BMP）为基础的骨组织工程技术是治疗颜面部骨缺损的研究热点之一。同行研究表明：因为供体表观遗传修饰差异极大，hASCs对于BMP的成骨诱导具有反应性低和不确定性。改变hASCs的成脂分化趋势，重置成骨分化通路是提高hASCs对于BMP成骨诱导反应性的关键。我们前期研究发现通过antimiR-34a这一表观遗传修饰可以显著增强hASCs对于BMP成骨诱导关键蛋白的表达，因此具有良好的应用前景。然而，antimiR-34a促进hASCs成骨分化的分子机制未见报道。因此提出假设：antimiR-34a通过上调Notch-TAZ、NF-κB-TAZ通路和直接作用于BMP通路是实现其促进hASCs成骨分化的主要机制。在本研究中，我们拟阐明antimiR-34a促进hASCs成骨分化的信号通路机制，为构建以hASCs和BMP为基础的组织工程化骨提供理论基础。

以脂肪干细胞(hASCs)和骨形成蛋白(BMP)为基础的骨组织工程技术是治疗颜面部骨缺损的研究热点之一。同行研究表明hASCs对于BMP的成骨诱导具有反应性低和不确定性。改变hASCs的成脂分化趋势，重置成骨分化通路是提高hASCs对于BMP成骨诱导反应性的关键。我们前期研究发现通过antimiR-34a这一表观遗传修饰可以显著增强hASCs对于BMP成骨诱导关键蛋白的表达，但antimiR-34a促进hASCs成骨分化的分子机制仍不明确。本次课题研究证实NICD-TAZ、SMAD1/5-TAZ及NF-κB-TAZ通路参与 antimiR-34a诱导hASCs 成骨分化，并且antimiR-34a与BMP能协同诱导 hASCs的成骨分化。