(174) ZNF408/progesterone signaling pathway regulates decidualization of human uterine stromal cells by regulating angiogenesis

Ran Tian, Liyan Wu, Ziwei Ai, Yueyue Hu, Ying Zhang,  Honglu Diao^*

Biomedical Engineering College; Reproductive medicine center, Renmin Hospital; Hubei University of Medicine, Shiyan, Hubei 442000, P. R. China

Abstract Text:

Recurrent Spontaneous Abortion (RSA) is a relatively common disease in obstetrics and gynecology clinics, in which impaired decidualization is one of the main causes of recurrent spontaneous abortion. It has been shown that ZNF408 knockout results in severe radial vascular defects in the zebrafish retina, accompanied by abnormal trunk vascularization. Whether ZNF408 regulates angiogenesis during endometrial decidualization in patients with RSA and its specific mechanism are not clear. Mice with recurrent spontaneous abortions were established in vivo. In vitro, human endometrial stromal cells were treated with MPA and cAMP to induce decidualization. Quantitative real-time PCR and Western blotting were used to detect ecdysis markers and important signaling molecules during ecdysis. Our study found that ZNF408 expression was steadily elevated during decidualization. Uterine vasculature was impaired in RSA mice and ZNF408 expression was significantly downregulated in RSA patients and mice. Knockdown of ZNF408 in uterine stromal cells for RNA sequencing significantly inhibited progesterone receptor PGR expression. Notably, the expression of ZNF408 was gradually decreased under the induction of CAMP alone and increased under the induction of MPA alone. The expression of ZNF408 could be inhibited by using RU486 (a progesterone receptor inhibitor). Thus，we proposed that there might be a possible interaction between ZNF408 and the progesterone receptor PGR, which was verified by the CO-IP assay. In addition, GO analysis showed that knockdown of ZNF408 significantly affected biological processes such as epithelial tube morphogenesis. In vascular endothelial cells, ZNF408 recombinant protein significantly enhanced the number and length of tube-forming loops promoting endothelial cells. VEGFA is involved in vascular recasting at the maternal-fetal interface as a major angiogenic and vascular permeability-inducing factor. ZNF408 co-localizes with VEGFA expression in uterine stromal cells. In this study, we focused on the potential mechanisms by which the ZNF408 /progesterone signaling pathway regulates angiogenesis during decidualization of the uterus in recurrent spontaneous abortions, which may provide some new therapeutic strategies for the treatment of recurrent spontaneous abortions due to impaired vascular function.  
*Correspondence: Honglu Diao, Email: hldiao@hbmu.edu.cn