基于右室双出口型先天性心脏病的差异表达lncRNA筛选及作用机制探讨

Congenital heart disease (CHD) is the most common birth defects in China, the major causes are usually considered the interactions between environmental factors and intracellular genetic materials. Long non-coding RNA（lncRNA）is one of the most important epigenetic mechanism. Recently, growing evidence has confirmed that lncRNA served as new regulators in cardiac development. At present, the research of lncRNA in CHD, there were only few studies in septal defect. In our previous study, through global genome-wide DNA sequencing and methylation sequencing, RNA sequencing, we found differently expressed mRNA which are related to cardiac development, but the mechanism is unknown, maybe due to the other epigenetic mechanism. In this study, we will survey the differentially expressed lncRNA and mRNA in cardiac tissue of monozygotic twin discordant for cardiac double outlet right ventricle (DORV) by lncRNA microarray. Through bioinformatics analysis, we will screen the differentially expressed lncRNA which target gene is associated with cardiac development. Then, we will perform the confirmation experiment in the case-control samples using real-time fluorescent quantitative polymerase chain reaction（qRT-PCR）method with TaqMan probe, and identify the DORV relevant lncRNA. This study will reveal the epigenetic mechanism that lncRNA regulate cardiac development related gene expression in cardiac double outlet right ventricle, and provide scientific basis for further exploration of the cause, mechanism and early prevention of CHD.

先天性心脏病（先心病）是我国最常见的出生缺陷，主要病因是环境和遗传因素交互作用即表观遗传。研究表明作为表观遗传方式之一的长链非编码RNA（lncRNA）在心脏发育过程中具有潜在作用。目前，lncRNA仅在间隔缺损先心病中有零星报导。我们前期对心脏右室双出口表型差异同卵双胎心脏组织进行全基因组的DNA测序和甲基化测序、及RNA测序，发现与心脏发育相关基因的差异表达，原因尚不清楚，推测其它表观遗传机制。因此，本项目拟采用lncRNA芯片，对双胎心脏组织进行差异表达lncRNA和mRNA的筛选；进行生物信息学分析，确定靶基因为心脏发育相关的lncRNA，通过qRT-PCR在病例-对照样本中对lncRNA及其靶基因的mRNA进行验证，确定目标lncRNA。本研究将揭示lncRNA通过调控心脏发育相关基因的mRNA表达而引起先心病发生的表观遗传机制，结果将为进一步探索先心病因和防治策略提供理论指导。

先天性心脏病（先心病）是我国最常见的出生缺陷，主要病因是环境和遗传因素交互作用即表观遗传。研究表明作为表观遗传方式之一的长链非编码RNA（lncRNA）在心脏发育过程中具有潜在作用。目前，lncRNA仅在间隔缺损先心病中有零星报导。我们前期对心脏右室双出口表型差异同卵双胎心脏组织进行全基因组的DNA测序和甲基化测序、及RNA测序，发现与心脏发育相关基因的差异表达，原因尚不清楚，推测其它表观遗传机制。在本研究中，采用RNA-Seq技术，对右室双出口表型差异同卵双胎心脏组织进行RNA测序，然后筛选差异表达的lncRNA和mRNA；进行生物信息学分析，确定靶基因为心脏发育相关的lncRNA，筛选出前13位的lncRNA-心脏发育相关mRNA的关系对；通过qRT-PCR在散发的病例-对照样本中进行验证，没有发现有统计学差异。本研究的实验结果尚不能提出lncRNA通过调控心脏发育相关基因的表达而引起CHD的表观遗传机制，但是本研究首次绘制了右室双出口表型差异同卵双胎心脏组织的lncRNA图谱和mRNA转录组图谱，为后续的分析与探索研究提供了基础测序数据。