microRNA-30下调DOC2A基因促进肠神经发育:先天性巨结肠治疗新策略?

Hirschsprung's disease(congenital megacolon) is one of the most common pediatric gastrointestinal malformation.The fundamental cause of HD is enteric nervous system developmental disorder.The applicant's previous studies have confirmed that microRNAs are essential for neuronal development and DOC2A gene konckdown promotes neural development and migration, and microRNA-30 is predicted to regulate DOC2A gene via the bioinformatics. In light of these,we inversely deduced the scientific hypothesis that microRNA-30→ downregulates DOC2A gene →promotes enteric nervous system development.Our project, based on our previous studies, will focus on microRNA-30 downregulating DOC2A gene, and study the molecular mechnism underlying microRNA-30 promoting enteric neural development and enteric neural crest cell migration in vitro and in vivo to confirm the scientific hypothesis: microRNA-30→downregulates DOC2A gene→promotes enteric neural development and enteric crest cell migration. Thus,our project will lay the foundation for the theory that microRNA-30 promotes enteric neural development and further explore the novel molecular treatment strategy for Hirschsprung's disease.

先天性巨结肠是最常见的小儿消化道发育畸形之一。目前明确其根本病因是肠神经发育障碍。申请人的前期研究已经证实microRNA在神经发育中发挥关键调控作用，同时申请人另一项前期研究证实下调DOC2A基因显著促进神经发育、成熟和迁移，据此研究基础，并进一步通过生物信息学方法预测microRNA-30下调DOC2A基因，从而逆推导出microRNA-30→下调DOC2A基因→促进神经发育的科学假设。本课题基于申请人前期研究的基础上，拟通过研究microRNA-30下调DOC2A基因，并从体内和体外两方面实验阐明microRNA-30促进肠神经发育和肠神经嵴细胞迁移的分子机理，从而验证microRNA-30→下调DOC2A基因→促进肠神经发育和肠神经嵴细胞迁移的科学假说，继而为应用microRNA-30促进先天性巨结肠肠神经发育奠定理论基础，并探索新的分子治疗策略。

先天性巨结肠是最常见的消化道发育畸形之一。本课题通过研究证实microRNA-30d下调其下游基因DOC2A的表达,并进一步研究证明microRNA-30d促进肠神经发育,研究证实microRNA-30d促进肠神经嵴细胞迁移。本课题进一步开展拓展研究，通过收集罕见的先天性巨结肠家系进行全外显子测序，发现新的致病基因，并应用CRISPR/Cas9成功构建斑马鱼和小鼠基因敲除模型，并进一步解析新致病基因的分子机制。本课题为应用非编码RNA治疗先天性巨结肠探索新策略，并拓展发现了先天性巨结肠新致病基因并解析其分子机制。研究成果：发表高水平SCI论文5篇，中文论文1篇；培养硕士研究生两名。