环状RNA Cir-coro1c调控CORO1C影响肠神经迁移在先天性巨结肠发生中的机制研究

Hirschsprung’s disease (HSCR) is one of the common congenital digestive disorder with uncertain mechanism. Recently, none coding RNAs seemed to be involved in the process of HSCR by epigenetics studies. By circular RNA chip array, cir-coro1c showed to be most stable with significant difference and was confirmed to be high expressed in HSCR tissues. Meantime, quantitative real-time PCR were applied to detection expression of cir-coro1c linear RNA coro1c in HSCR samples and found that coro1c showed lower expression level in HSCR tissues and had a significant negative correlation with cir-coro1c. ShRNA-mediated knock-down of cir-coro1c promoted the cell migration with a high regulated coro1c mRNA in SH-SY5Y cell lines. Thus, we hypothesized that cir-coro1c might affect neural crest cell migration by regulating coro1c in the pathogenesis of HSCR. In the present proposal, employing human sample, cell and molecule, we will fully investigate the roles of cir-coro1c and coro1c in the defect of neural crest cell migration, which will offer new insights into the pathogenesis and embryonic period prevention of HSCR.

先天性巨结肠是常见的消化道畸形之一，目前具体发病机制尚未明确。近年来，表观遗传学研究表明，非编码RNA在先天性巨结肠的发生、发展中起着重要作用。我们前期通过环状RNA芯片筛选，发现cir-coro1c表达差异最稳定且显著，进一步人群样本验证发现cir-coro1c呈与芯片一致的显著高表达。而其对应线性RNA coro1c显著低表达并与cir-coro1c表达呈显著负相关关系，同时体外实验证实cir-coro1c低表达可促进神经细胞迁移能力，并且伴随着coro1c mRNA表达水平上调。故推测：cir-coro1c可能通过调控coro1c表达，影响神经嵴细胞的迁移，与先天性巨结肠发生相关。本课题将在人群、体外细胞及分子水平深入研究cir-coro1c和coro1c影响肠神经嵴细胞迁移障碍的作用，为先天性巨结肠的发病机制及胚胎期防治提供新视点。

肠神经嵴细胞 (ENCCs)迁移和增殖障碍导致消化道远端神经节细胞缺如是先天性巨结肠（HSCR）发病的主要原因，目前具体发病机制尚未明确。本课题主要研究环状RNA（cirRNA）及其宿主基因在HSCR发生中的作用机制。.我们用cirRNA芯片筛选HSCR病变段差异表达cirRNA时发现cir-coro1c显著高表达。生物信息学显示coro1c是cir-coro1c的宿主基因。人群验证发现cir-coro1c水平高表达同时coro1c低表达。体外实验提示cir-coro1c高表达能够抑制细胞的增殖、迁移功能，而coro1c的功能与之相反。高表达的cir-coro1c能够抑制coro1c的表达。模拟体内环状RNA水平，高表达的cir-coro1c可以通过影响coro1c的mRNA稳定性，并且造成CORO1C与细胞骨架相关蛋白Rac1和ARP2/3的结合下降，导致细胞迁移受损。.本研究首次阐述cir-coro1c和宿主基因coro1c的调控关系及其在先天性巨结肠发生中影响神经嵴细胞迁移和增殖功能，为先天性巨结肠的发生提供新的研究方向，为先天性巨结肠的发病机制及胚胎期防治提供新视点。