增强子RNA介导炎症反应在不明原因复发性流产中的作用及其分子机制

Unexplained recurrent spontaneous abortion (URSA) is a kind of pathological pregnancy, the etiology and pathogenesis of which is the main research hotspots in the field of reproduction. Accumulating evidences have demonstrated that the enhancer RNA (eRNA), a special class of long non-coding RNA (lncRNA), has received growing attention of the roles in the occurrence and development of kind of diseases. According to our previous studies, we have proposed that eRNA may mediate inflammatory pathways through recruitment of trans-acting factors, and then leads to the occurrence of URSA. Based on our established prospective birth cohort (NJMU Birth Cohort), we will start with transcriptome sequencing of chorionic villus tissue, and apply luciferase reporter assay to screen and identify the potentially functional eRNA associated with URSA. Besides, through functional studies in animal models and in vitro experiment, we will systematically access the effect of eRNA on embryonic development, clarify the role of eRNA mediating the inflammatory pathway in the occurrence of URSA, and reveal potential mechanisms of action of eRNAs in the regulation of embryo development. This project will open avenues for the diagnosis and treatment of URSA, improve the assisted reproductive technology risk assessment and clinical standards, and make contributions to improve the quality of the population and promote reproductive health.

不明原因复发性流产(URSA)是一种病理性妊娠，其病因和发病机制一直是生殖领域研究的热点。近年研究发现，增强子RNA (eRNA) 作为一类特殊的长链非编码RNA (lncRNA)，其在疾病发生发展过程中有十分重要的作用。我们的前期研究结果提示eRNA可能通过招募反式作用因子介导炎症通路，进而导致URSA的发生。本项目拟基于已建立的大样本前瞻性出生队列，针对流产绒毛组织，应用转录组测序及独立人群验证，筛选与URSA相关的eRNA，通过荧光素酶报告基因、ChIP-qPCR等实验鉴定具有潜在功能的eRNA；并结合体外细胞实验及整体动物模型，系统论证eRNA对胚胎发育的影响，阐明eRNA介导炎症通路在URSA发生中的作用与分子机制。本项目研究可为URSA的分子诊断和治疗提供新的途径，同时完善辅助生殖技术风险评估及临床标准，为提高人口质量、促进生殖健康做出贡献。

增强子RNA (eRNA) 作为一类特殊的长链非编码RNA (lncRNA)，在不明原因复发性流产（URPL）发生发展过程中起到重要作用。本项目基于已建立的大样本前瞻性出生队列，针对绒毛、蜕膜组织，采用RNA-seq高通量测序、荧光素酶报告基因、ChIP-qPCR等筛选和鉴定与URPL相关的eRNA，进而结合体外细胞实验及整体动物模型，揭示其可能机制。结果显示，URPL患者绒毛组织中lnc-SLC4A1-1高表达；斑马鱼胚胎显微注射实验提示，过表达eRNA可以影响斑马鱼胚胎发育；细胞功能实验提示其可抑制细胞增殖促进滋养层细胞的迁移和凋亡。机制研究发现，lnc-SLC4A1-1通过募集转录因子NF-κB与CXCL8结合，使CXCL8启动子区H3K27ac增加，激活CXCL8转录，诱导TNF-α和IL-1β水平上升，加剧滋养细胞的炎症反应。在URPL患者蜕膜组织中，我们鉴定出eRNA lnc-CES1-1，其在蜕膜组织可与下游基因PPARγ相互作用，影响蜕膜滋养层细胞迁移，参与URPL的发生。在此基础上，申请并获得授权“用于诊断不明原因复发性流产的增强子RNA组合、引物组及应用和试剂盒”（专利号：ZL 201710735738.6）国家发明专利。本研究为了解lncRNA/eRNA在病理性妊娠中的功能提供了新的理解，为URPL分子诊断和治疗提供新的途径。