长链非编码RNA在第一二腮弓发育及相关综合征发生中的作用

The first and two branchial arch syndrome is a craniofacial abnormalities caused by neural crest malformations. Its incidence is only after cleft lip and palate. The mechanism is not entirely clear yet. Long non-coding RNA (lncRNA) is a class of RNA which does not encode proteins in vivo .lncRNA can regulate gene expression from the epigenetic, transcriptional regulation, and other aspects of transcriptional regulation and may become possible biomarkers .In preliminary work we used retinoic acid to intervent chondrocyte cells derived from neural crest and analyses changes of the whole genome expression. results showed that the expression directly generate of lncRNA RNA polymerase II which directly generate lncRNA is significantly increased. suggesting lncRNA may be involved in the occurrence of deformities as a bridge between environmental and genetic factors. This study intends to use the pre-established rat model with malformation of the first and two branchial arch induced by retinoic acid and high-throughput sequencing technology to study the expression of lncRNA in different developmental periods of the first and two branchial arch and compare the expression of lncRNA in normal and abnormal development of the first and two branchial arch. Results of these studies will be used to establish expression patterns of lncRNA in different developmental stages of the first and two branchial arch and filter out iconic lncRNAs for further studies. These studies will clear the role of lncRNA in the development of the first and second branchial arch and the occurrence of the first and two branchial arch syndrome, so as to provide new clues to pathogenesis of the first and two branchial arch syndrome.

第一二腮弓综合征是颅面部神经嵴组织发育异常所致畸形，发病率仅次于先天性唇腭裂畸形，发生机制尚不完全明确。长链非编码RNA(lncRNA)是生物体内不编码蛋白质的一类RNA，可从表观遗传学、转录调控及转录后调控等多种层面实现对基因表达的调控，并可作为生物标志物。前期工作中利用维甲酸干预神经嵴来源软骨细胞对基因组表达分析发现直接生成lncRNA的RNA聚合酶II的表达被明显上调，提示lncRNA作为环境因素与遗传之间的桥梁参与了畸形的发生。本研究拟利用前期建立的维甲酸干预致第一二腮弓发育异常大鼠模型，采用高通量测序技术对lncRNA在不同时段胚胎第一二腮弓正常及异常发育的表达进行对比研究，确定lncRNA在第一二腮弓发育不同阶段的表达模式，并筛选出标志性lncRNA进一步研究，以明确lncRNA在第一二腮弓发育及第一二腮弓综合征发生中的作用，从而为第一二腮弓综合征发病机制的研究提供新的线索。

背景：第一二腮弓综合征是颅面部神经嵴组织发育异常所致畸形，发病率仅次于先天性唇腭裂畸形，发生机制尚不完全明确。长链非编码RNA(lncRNA)是生物体内不编码蛋白质的一类RNA，可从表观遗传学、转录调控及转录后调控等多种层面实现对基因表达的调控，并可作为生物标志物。.主要研究内容：利用维甲酸干预特定时期大鼠胚胎发育，制作第一二腮弓综合征大鼠模型，对所得大体标本进行相关指标测量，对所得组织标本利用lncRNA芯片进行差异基因和差异lncRNA筛选，将获得的基因芯片数据利用String、Phenolyzer等生物信息学软件进行分析，重点分析差异基因之间的联系，观察这些基因之间的相互作用，筛选出居于相互作用网络中关键地位的基因。.重要结果：G10天干预对胚胎整体发育影响较小，但对颅面部发育影响较大，表现为鼻枕距及鼻耳距缩小，顶耳距增大，外耳位置整体向下及向背部移位，耳长、耳宽及耳指数均减少，不仅在外耳大小而且在耳廓位置及头颅整体发育上均与第一二腮弓综合征临床表现相符。利用T检验的p值和倍数变化值进行差异基因和差异lncRNA筛选，筛选的标准为上调或者下调倍数变化值>= 2.0且P值<= 0.05。共筛选出表达上调的mRNA5 17个，下调415个，lncRNA上调231个，下调172个（表5）。其中已知与第一二腮弓综合征畸形发病相关的基因包括six1、pax8、bmp5、emx2、eya1、tcf15、irf6、mesp1、fas、fgf10、hoxb3等，并利用相关生物信息学软件进行进一步分析，发现致病基因之间存在集群效应。.科学意义：通过对维甲酸诱导的第一二腮弓综合征大鼠模型组织标本行lncRNA芯片检测，获得了相应的表达变化的lncRNA，生物信息学分析发现畸形的发生可能与多个基因组成的基因网络整体功能受影响有关。