完全性葡萄胎血管发生障碍中外泌小体的影响及机制

项目名称： 完全性葡萄胎血管发生障碍中外泌小体的影响及机制

项目编号： No.81501255

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 赵九如

作者单位： 上海交通大学

项目金额： 18万元

中文摘要： 完全性葡萄胎(CHM)绒毛发育早期存在少量的血管生成，但因发育障碍未能成熟。外泌小体是胞外重要的miRNA载体，能够主动分选包装miRNA。绒毛血管来源于间充质干细胞（MSCs），而MSCs的分化严格依赖于微环境。我们的前期研究表明，与正常绒毛间质细胞培养上清外泌小体相比CHM囊液中的外泌小体能够显著抑制原代脐静脉内皮细胞（HUVEC）的血管形成能力。故本项目拟在分离CHM囊液、CHM患者血清以及正常绒毛间充质培养上清中的外泌小体基础上，分别进行miRNA测序和质谱实验，筛选出差异性的血管生成相关蛋白和miRNA。进而分别在MSCs和HUVEC上研究差异蛋白和miRNA对其向血管分化、发育的影响，并进一步对差异miRNA参与的信号通路及靶基因进行预测和验证。本项目不仅可以深入揭示CHM血管发生障碍的机理，也为其他存在绒毛血管发育异常的疾病如FGR、PIH等发病机制提供有力的理论指导。

中文关键词： 胎盘血管；完全性葡萄胎；外泌小体；miRNA；间充质干细胞

英文摘要： Complete hydatidiform mole (CHM) is one of the main gestational trophoblastic diseases. CHM is the product of an abnormal pregnancy with swollen chorionic villi and hyperplasic trophoblasts, but without mature vessels. However, immuno-fluorescence with CD34 and other vessels markers prove, there does exist vasculogenesis during the early villi development stage, but unable to mature for defects in further development. The villious vessels are generally believed differentiated from mesenchymal stem cells (MSCs), while the differentiation processes of MSCs are regulated by numerous molecules with complex signal pathway at different levels. Moreover, the differentiation of MSCs is dependent on the microenvironment they located. Exosomes are small (40–100 nm) membrane vesicles of endocytic origin that are released into the extracellular environment and are involved in cell-to-cell communication. Exosomes are important extracellular transporters of miRNAs, and they sorting miRNA automatically by hnRNPs and then functionally transferred to recipient cells. Our experiments with exsomes isolated from villous stroma cultures or CHM hydatidiform liquid proved the exosomes from CHM hydatidiform liquid inhibit the angiogenic sprouting and capillary lumen formation progress by human umbilical vein endothelial cells (HUVEC).In this project, we plan to isolate exosomes from CHM hydatidiform liquid, CHM patient serum and villous stroma cultures. Using PKH67 labeling, we would observe the dynamic process of exosomes absorption by MSCs and HUVEC. After miRNA high-throughout sequencing and protein mass spectrometry sequencing, we would isolate the angiogenesis and vasculogenesis associated differential expressed proteins and miRNAs. Next, we will construct the lenti-viral vectors of those proteins and infect MSCs or HUVEC to study the effects on the differentiation to vessels of MSCs and effects on the capillary lumen formation of HUVEC. Besides, we will package the differential miRNA mimics to exosomes and further to study their effects on MSCs or HUVEC. Finally, we will investigate the pathways or targets of those miRNAs involved in. Our findings will be important for revealing the mechanism of vessels development defects in CHM and rich scientific theory for other trophoblastic diseases with villious abnormality, such as FGR and PIH.

英文关键词： Placental vessel;Complete Hydatidiform Mole;Exosomes;miRNA;Mesenchymal Stem Cells