非编码RNA调控维生素A缺乏致先天性脊柱侧凸的作用及机制研究

The sophisticated interaction of environmental and genetic factors is thought to be the pathogenesis resulting congenital scoliosis(CS). Environmental factor in embryo may play an important role in the onset of CS, but the mechanism was unclear. Vitamin A deficiency(VAD) during pregnant theoretically may cause CS.In a previous study, we have found that vitamin A deficiency can induce congenital scoliosis. However, the molecular mechanism underlining its pathogenes is not well known. We used miRNA-Chip and lncRNA-chip to analysis the miRNA and lncRNA which were were significantly higher or lower in CS than the normal one. Therefore, the techniques of siRNA gene knock-in and gene knock-out will be used to detect the mechanism of appearance between the CS and the normal one in this study. And verification of the difference will be conducted through the patients. According to individual-group and globe-cell-animal model studies, we plan to find the related noncoding RNA for the genesis and prognosis of CS. This study will give a new insight into the pathogenesis of CS and provide a new way for the early diagnosis, treatment and prevention of CS.

先天性脊柱侧凸(CS)的诊治是脊柱外科的难题，其发生除与遗传因素相关外,环境因素也发挥重要作用，但具体病因和机制尚不明确。流行病学和发育学研究提示维生素A缺乏可能在CS发生发展过程中起重要作用。前期的研究中，我们证实维生素A缺乏能导致CS的发生，但其具体机制尚不明确。我们推测非编码RNA可能调控维生素A缺乏致CS的发生发展。为证明此假说，本课题将应用miRNA芯片、lncRNA芯片筛选出调控维生素A缺乏致CS发生相关的miRNA、LncRNA；进一步应用siRNA、基因敲入、基因敲除等技术在细胞水平、动物模型中进行基因表达调控和功能的验证，从动物模型－环境因素－非编码RNA－病例多个层次，寻找和阐明调控维生素A缺乏致CS发病相关的分子机制。目前国内外尚未见CS环境因素－非编码RNA研究的报道，本研究将为CS的预防、早期诊治提供新的思路和新的靶点。

先天性脊柱侧凸(CS)是脊柱外科的诊疗难题之一，其发生除遗传因素相关外，环境因素也至关重要，但具体病因机制尚未阐明。流行病学和发育学研究提示维甲酸信号通路异常对CS发生、发展关系密切。前期研究中，我们发现孕期母鼠维生素A缺乏(VAD)可导致新生鼠CS的发生，推测非编码RNA可能参与调控VAD所致体节发育的异常，进而导致CS的发生。为证明此假说，本课题应用转录物组高通量测序，通过生物信息学分析筛选出差异表达的非编码RNA，构建ceRNA调控网络，进一步应用RT-PCR、荧光素酶报告系统、Northern Blot及Western Blot等技术在细胞水平及动物模型中进行功能学验证，从环境因素-动物模型-非编码RNA调控-临床表型多个层次，寻找和阐明调控维生素A缺乏致CS中体节发育异常相关的分子机制。目前国内外有关维生素A素缺乏-CS-非编码RNA调控的研究报道罕见，本研究将为CS的预防、早期诊治提供新的思路和靶点。