基于Jmjd6/sFlt1通路探讨E3泛素化连接酶HECTD1调控胎盘迷路区血管分化发育的机制

The placenta is an important organ for exchange of gas, nutrients and metabolite between maternal and fetal, and the well development of placental vasculature is the basis of healthy pregnancy. Hectd1 acts as an important E3 ubiquitin ligase, which plays a key role in physiological and pathological processes such as tumor occurrence, cardiovascular system, neurodevelopment and immune regulation. In the preliminary work of this project, loss of Hectd1 resulted in defects in mouse placental development and embryonic growth, and the levels of Jmjd6,VEGF, sFlt1 and other genes related to placental vascular differentiation showed significant changes. By further using the Hectd1 knockout mouse model, this project intends to observe the generation of villus branching, the changes of apoptosis/autophagy and angiogenesis-related proteins and receptors in the placental labyrinth. Taking advantage of techniques such as gene silencing and overexpression, co-immunoprecipitation, as well as gene sequencing, to confirm that Jmjd6 is a potential ubiquitination regulatory substrate for Hectd1, and to further clarify thatHectd1 specifically regulates the of villus branching and angiogenesis in the placental labyrinth through the Jmjd6 signaling cascade. This study will provide a new theoretical and experimental basis for understanding the pathological mechanism of fetal growth and development related diseases caused by placental angiogenesis disorders and exploring the diagnosis and treatment methods.

胎盘是母体/胎儿气体、营养物和代谢产物交换的重要部位，胎盘脉管系统的正常发育是良好妊娠的基础。Hectd1作为重要的E3泛素化连接酶，其在肿瘤、心血管系统、神经发育和免疫调节等生理病理过程起关键作用。本项目前期工作发现Hectd1缺失导致小鼠胎盘发育缺陷和胚胎生长发育受限，与胎盘血管分化相关的Jmjd6、VEGF、sFlt1等基因表达发生明显变化。此项目拟进一步利用Hectd1基因敲除小鼠模型，观察胎盘迷路区绒毛分支的生成、凋亡、自噬及血管形成相关蛋白及受体表达的变化。通过基因沉默和过表达、免疫共沉淀、基因测序等技术，证实Jmjd6为Hectd1潜在的泛素化调节底物，明确Hectd1通过Jmjd6级联信号通路特异性调节胎盘迷路区绒毛分支和血管形成的分子机制。该研究将为深入了解胎盘血管分化发育障碍导致的胎儿生长发育受限相关病理机制及探索诊治措施提供新的理论和实验依据。

胎盘是母体/胎儿气体、营养物和代谢产物交换的重要部位，胎盘脉管系统的正常发育是良好妊娠的基础。Hectd1作为重要的E3泛素化连接酶，其在肿瘤、心血管系统、神经发育和免疫调节等生理病理过程起关键作用。本项目前期工作发现Hectd1缺失导致小鼠胎盘发育缺陷和胚胎生长发育受限，与胎盘血管分化相关的Jmjd6、VEGF、sFlt1等基因表达发生明显变化。利用Hectd1基因敲除小鼠模型，观察胎盘迷路区绒毛分支的生成、凋亡、自噬及血管形成相关蛋白及受体表达的变化。证实了Hectd1对胚胎发育中胎盘迷路区的绒毛分支及血管形成的调节作用，通过体外细胞培养及体内实验证实了Hectd1通过Jmjd6/sFlt1通路作用于胎盘血管分化发育，通过免疫共沉淀、免疫组化等实验从体外、体内实验分层次逐步验证Hectd1基因对于胚胎和胎盘发育的重要性。通过基因沉默和过表达、免疫共沉淀、基因测序等技术，证实Jmjd6为Hectd1潜在的泛素化调节底物，明确Hectd1通过Jmjd6级联信号通路特异性调节胎盘迷路区绒毛分支和血管形成的分子机制。除Jmjd6外，实验发现Sirtuin1（乙酰化酶家族成员之一）也是Hectd1底物之一，补充实验验证Sirtuin1-NDRG1-p53通路，进一步论证 Hectd1 在胎盘发育中调节胎盘迷路区血管生成的分子机制，证实Hectd1-Jmjd6-U2AF65-sFlt1-VEGF/PIGF 通路。该研究将为深入了解胎盘血管分化发育障碍导致的胎儿生长发育受限相关病理机制及探索诊治措施提供了必要的理论和实验数据支持。