先天性脊柱侧凸（CS）是一种复杂的脊柱畸形，常合并脊髓纵裂（SCM），但是具体发病机制不明。研究提示CS体节发育异常与神经管关系密切，而DNA甲基化异常在神经管发育不良中起重要作用。申请人前期通过全基因组甲基化芯片及功能验证发现神经管发育BRE基因高甲基化而表达沉默，从而影响Wnt信号通路关键蛋白Wnt3a，Nkd1，β-catenin的表达。由此提出假说：高甲基化的BRE基因可能通过调控Wnt/β-catenin信号通路影响神经管发育进而干扰体节形成，在CS合并SCM的发生中起到关键作用。围绕上述假说，本课题拟采用甲基化特异性PCR、siRNA干扰、过表达等方法，在体外探讨甲基化修饰BRE基因调控Wnt/β-catenin信号通路的分子机制；并构建BRE基因敲除孕鼠模型，在体内分析其对胚胎体节和神经管发育的关键作用，以期为先天性脊柱脊髓畸形的预防、早期诊治提供新的思路和靶点。

Congenital scoliosis (CS) is a kind of complex spinal deformity, which often associated with split cord malformation (SCM). However, its etiology remains unknown. Neural tube defects can cause abnormal development of the somitogenesis which results in CS. Aberrant DNA methylation plays an important role in neural tube defects. In our previous study, the whole-genome methylation analysis indicated the hypermethylation in the promoter region of BRE gene in patients diagnosed with CS and coexisting SCM. Furthermore, functional verification experiments demonstrated misexpression of BRE could affect the expression of key proteins (Wnt3a, Nkd1, β-catenin) in Wnt signaling pathway. Thus, we hypothesized that hypermethylation of BRE may interfere with normal neural tube and somite development through Wnt/β-catenin signaling pathway, which may plays an important role in pathogenesis of CS with SCM. To prove this hypothesis, the techniques such as methylation-specific PCR and siRNA will be used to explore the molecular mechanism of methylation-modified BRE gene regulating Wnt/β-catenin signaling pathway in vitro; in vivo, BRE gene knock-out will be used to reveal its effect on somite and neural tube development. This study will give a new insight into the pathogenesis research and provide a new way for the early diagnosis, treatment and prevention of congenital spinal deformities and cord malformation.