构建3D人子宫内膜模型研究TNC与整合素介导的胚胎植入分子机制

胚胎植入及发育的关键因素包含高容受性子宫内膜、优质胚胎及同步的着床时间窗。但目前试管助孕条件下，第三天的胚胎植入子宫内膜的成功率仅为35%左右，且其中还有10%左右的胎停、流产率。为此我们前期利用人囊胚和原代子宫内膜细胞共培养体系，模拟了胚胎滋养细胞迁移/定位/粘附和侵入子宫内膜过程，成功观察到子宫内膜肌腱蛋白C（TNC）及滋养细胞整合素αvβ3高表达。本项目拟：构建3D人子宫内膜容受性研究模型；采用单细胞高通量测序，明确调控滋养细胞粘附/侵入的子宫内膜细胞类型，精准建立人胚胎植入窗口期子宫内膜容受性的表观遗传学分子调控网络；利用CRISPR/Cas9和受体阻断抗体等技术，解析子宫内膜的关键基因TNC（配体）结合滋养细胞整合素（受体）亚型，及影响滋养细胞粘附/侵入的相关决定因素。为进一步明确胚胎植入分子调控机制、寻找内膜容受性及胚胎种植窗标记物、探索反复种植失败及胎停流产原因奠定基础。

The key factors for embryo implantation and development include highly receptive endometrium, high quality embryos, and a synchronous implantation window. But under current IVF conditions, the success rate of embryo implantation is only about 35 percent on day 3, and about 10 percent of them are fetal arrest and miscarriage. Therefore, we used the co-culture system of human blastocyst and primary endometrial cells to simulate the process of embryo trophoblastic cell migration/localization/adhesion and invasion of endometrium, and successfully observed the high expression of endometrial tenascin C (TNC) and trophoblast integrin αvβ3. This project plans to construct a 3D human endometrial receptivity research model. Single-cell high-throughput sequencing is proposed to be used to determine the types of endometrial cells that regulate trophoblast cells adhesion/invasion, and precisely establish the epigenetic molecular regulation network of endometrial receptivity in the window of human embryo implantation. Planning to use techniques such as CRISPR/Cas9 and receptor blocking antibody to analyze the endometrial key gene TNC (ligand) binding to trophoblast integrin (receptor) subtype and related determinants affecting trophoblast adhesion/invasion. It will lay a foundation for further clarifying the molecular regulation mechanism of embryo implantation, searching for endometrial receptivity and embryo implantation window markers, and exploring the causes of repeated implantation failure and pregnancy loss.