小鼠子宫内膜胚胎着床点CircularRNA的表达及其对胚胎着床的调控作用

Embryo implantation is one of the key links in the pregnancy success, but the mechanism has not yet been elucidated. Studies have shown that CircRNA is closely related to a variety of physiological and pathological process. It can act as competitive endogenous RNA combined with miRNA, then remove the miRNA inhibition to its target genes and increase the expression of target genes. In embryo implantation, the function of miRNAs was systematically studied in our preliminary work, but its upstream circRNA function in embryo implantation and establishment of endometrial receptivity are unclear at present. This project aims to detect circRNA expression of decidual tissue from abortion patients and normal pregnancy as well as endometrial tissue of implantation site and inter-implantation site in the mouse by circRNA chip, in order to screen relevant CircRNA of the endometrial receptivity establishment. At the same time, combined with the our previous work of miRNA chip data and gene chip data, the downstream target miRNA - mRNA of CircRNA regulation will be predicted by bioinformatics according to the "CircRNA - micrornas - mRNA" train of thought, and further confirm its role and regulation mechanism in receptivity establishment through a series of functional trials in vivo and in vitro. This project will have a great clinical value in the diagnosis and treatment of infertility caused by Implantation failure and the improvement of assited reproduction.

胚胎着床是妊娠成功与否的关键环节之一，机制尚未阐明。研究显示CircRNA与多种生理病理过程密切相关，可充当竞争性内源RNA与miRNA结合，进而解除miRNA对其靶基因的抑制，上调靶基因表达。课题组前期系统研究了miRNA在胚胎着床过程中的作用，但其上游CircRNA在胚胎着床子宫内膜容受性建立这一过程中的作用尚未有研究报道。本研究拟对临床流产患者、正常妊娠者蜕膜组织以及妊娠小鼠子宫内膜胚胎着床点及其旁组织进行CircRNA芯片检测，筛选子宫内膜容受性相关CircRNA，同时结合课题组前期miRNA芯片数据和基因芯片数据，按照“CircRNA-miRNA-mRNA”的思路，对CircRNA调控的下游靶miRNA-mRNA进行预测和验证，并通过一系列体内外功能实验证实CircRNA在容受性建立中的作用及调控机制。本项目将对着床失败所致不孕症的诊断和治疗以及新的生殖调控手段的开发有重要意义。

胚胎着床是妊娠成功的关键环节之一，机制尚未阐明。研究显示CircRNA与多种生理病理过程密切相关，可充当竞争性内源RNA与miRNA结合，解除miRNA对其靶基因的抑制。课题组前期研究了miRNA在胚胎着床过程中的作用，但其上游CircRNA在胚胎着床过程中的作用鲜有报道。本项目对临床流产患者、正常妊娠者子宫蜕膜组织以及妊娠小鼠子宫内膜胚胎着床点及其旁组织进行CircRNA芯片检测，筛选胚胎着床相关CircRNA，同时结合前期miRNA芯片数据和基因芯片数据，对CircRNA调控的下游靶miRNA-mRNA进行预测和验证，并通过一系列体内外功能实验探讨其在胚胎着床过程中的可能作用。通过研究，我们发现：.1、微阵列芯片成功筛选出早期不明原因复发性流产患者子宫蜕膜组织中123个差异表达的circRNAs，其中包括78个上调和45个下调的circRNAs；并对差异表达显著的12个circRNAs进行了验证。并通过生物信息学分析进一步构建了circRNA-miRNA-mRNA作用网络，包括3个 circRNAs (hsa_circRNA_104179、hsa_circRNA_103092、 hsa_circRNA_103093)、 27 个microRNAs (miR-224、miR-522-3p、 hsa-miR-421等)和 82个 mRNAs (H2AFV、 DIMT1、 PRLR等)。最后以hsa_circRNA_103092-miR-224-PRLR轴为目标，探讨其在早期不明原因复发性流产中的可能作用，初步发现hsa_circRNA_103092可能通过miR-224调控PRLR表达，调节细胞增殖。.2、运用Arraystar circRNA芯片筛选孕第5天小鼠子宫内膜胚胎着床点及其旁组织中差异表达的circRNAs，并对差异表达的circRNAs进行验证。生物信息学分析进一步构建了circRNA-miRNA-mRNA作用网络，并以circ-00888-miR-30c-RBPJ轴为目标，初步探讨其在早孕小鼠胚胎着床中的作用。研究发现circ-00888、miR-30c、RBPJ均在早孕小鼠子宫内膜中呈现规律表达，circ-00888可能通过对miR-30c的吸附，解除其对RBPJ的表达，进而参与调节子宫内膜蜕膜化。.通过本项目的资助，我们已公开发表5篇SCI论文。