神经嵴干细胞结合基因编辑对巨结肠模型鼠的移植治疗探索

项目名称： 神经嵴干细胞结合基因编辑对巨结肠模型鼠的移植治疗探索

项目编号： No.81671531

项目类型： 面上项目

立项/批准年度： 2017

项目学科： 医药、卫生

项目作者： 钭金法

作者单位： 浙江大学

项目金额： 25万元

中文摘要： 先天性巨结肠(HD)又称肠无神经节细胞症，是小儿外科常见急重症，是由于发育过程中肠神经嵴干细胞（NCSC）或重要基因的问题，导致NCSC在移行、分化、发育的过程中发生缺陷所致，一般由环境或遗传因素所导致。目前HD只能靠外科手术治疗，并有很多潜在并发症。为模拟临床问题，基因缺陷型和获得性HD模型得到了建立。干细胞的研究进展，为应用该技术治疗HD带来新的可能治疗途径。本课题组利用诱导多能干细胞诱导出纯化的神经前体细胞或类神经嵴前体细胞，并进一步分化出具有电生理活性的感觉神经细胞，在体外移植后得到了Synaptophysin以及特征性神经元nNOS的标记；其次已获取并培养出典型特征的肠NCSC。在两种神经嵴干细胞的基础上，为进一步解决先天遗传性问题，设计结合基因编辑，尝试解决EDNRB缺陷型小鼠的HD疾病。该课题的设计和完成，有望对HD疾病的治疗进行良好的探索，产生好的体内数据甚至再生修复。

中文关键词： 神经嵴干细胞；无神经节细胞症；移植；基因编辑

英文摘要： Hirschsprung's disease（HD）is a common neuron disease in pediatric surgery，also called aganglionsis. It is caused by the developmental failure of the enteric neural system （ENS）progenitors during migration into the gastrointestinal tract, particularly the distal colon. To this day, HD is only treated by surgical procedure with many possible complications. To mimic the HD in research, two HD model were established based on genetic and environmental elements individually. Stem cell is promising for HD therapy. To explore the treatment for HD, we isolate the pure sensory neural progenitors or NCSC-like progenitors from ESCs or IPS first, then differentiated into functionally active sensory neuron in vitro, and showed the evidence of nNOS and Synaptophysin expression when we transplanted these progenitors into enteric explants after 7 days. Another candidate NCSCs from postnatal mice were also isolated and proliferated with differentiation and electric physiology evidence as neuron. Genome editing combined with both candidate cells were proposed to explore the potential to cure genetic HD in vivo, specially on the EDNRB(-) HD model. Our proposal and research will explore new strategy for HD remedy, get some positive result in vivo, and function regeneration in cells and gut possibly.

英文关键词： neural crest stem cell; aganglionsis;transplatation;genome editing