EHD1 通过调节 Wnt4/β-连环蛋白信号通路在复发性植入失败中损害蜕膜化。
EHD1 impairs decidualization by regulating the Wnt4/β-catenin signaling pathway in recurrent implantation failure.
机构信息
Reproductive Medicine Center, The Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing 210008, People's Republic of China; State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences , Nanjing University , Nanjing 210023, People's Republic of China.
Reproductive Medicine Center, The Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing 210008, People's Republic of China.
出版信息
EBioMedicine. 2019 Dec;50:343-354. doi: 10.1016/j.ebiom.2019.10.018. Epub 2019 Nov 6.
BACKGROUND
Recurrent implantation failure (RIF) remains a critical and challenging problem in assisted reproductive technology mainly due to impaired decidualization. The endocytic and transcytotic activity in the endometrium are crucial for decidualization. The most representative endocytic gene is the C-terminal Eps15 homology domain-containing 1 (EHD1), but whether EHD1-mediated endocytic function is responsible for embryo implantation during decidualization remains unclear.
METHODS
A transcriptomic analysis was performed to evaluate the differentially expressed genes between the fertile control and RIF group. The expression and location of EHD1 in endometrial tissues were further examined by IHC, qRT-PCR and Western blotting. The transduction of an EHD1 recombinant adenovirus into human endometrial stromal cells was performed to investigate relevant decidualization marker genes. Additionally, a microarray analysis following the adenovirus-mediated overexpression of EHD1 was conducted to identify EHD1-related changes in HESCs, and the potential molecular mechanisms were further confirmed through immunofluorescence and coimmunoprecipitation analyses.
FINDINGS
An RNA-seq analysis demonstrated that EHD1 expression was significantly higher in the mid-secretory endometrium of the RIF group than in that of the fertile control group. The analysis of the menstrual cycle showed that expression of EHD1 increased in the mid-proliferative phase and showed a gradual decrease in the mid-secretory and decidual phases. Furthermore, EHD1 overexpression impaired decidualization by suppressing the expression of prolactin and insulin-like growth factor binding protein-1 and the formation of the cytoskeleton. The mechanistic analysis revealed the EHD1 regulated LRP5/6 protein function through the endocytic pathway, and subsequently suppressed the Wnt4/β-catenin pathway during decidualization. In addition, a Wnt4 agonist improved an impaired decidualization process.
INTERPRETATION
Regulation of the EHD1-Wnt4 pathway might serve as a promising therapeutic strategy for improving endometrial receptivity in RIF women.
背景
复发性植入失败（RIF）仍然是辅助生殖技术中的一个关键和具有挑战性的问题，主要是由于蜕膜化受损。子宫内膜的内吞和转胞作用对于蜕膜化至关重要。最具代表性的内吞基因是 C 末端 Eps15 同源结构域包含蛋白 1（EHD1），但 EHD1 介导的内吞功能是否负责胚胎在蜕膜化过程中的植入仍不清楚。
方法
通过转录组分析评估了肥沃对照组和 RIF 组之间差异表达的基因。进一步通过免疫组化、qRT-PCR 和 Western blot 检测 EHD1 在子宫内膜组织中的表达和位置。通过将 EHD1 重组腺病毒转导到人子宫内膜基质细胞中，研究相关的蜕膜化标记基因。此外，通过腺病毒介导的 EHD1 过表达进行微阵列分析，以鉴定 HESCs 中与 EHD1 相关的变化，并通过免疫荧光和共免疫沉淀分析进一步证实潜在的分子机制。
发现
RNA-seq 分析表明，RIF 组的中分泌期子宫内膜中 EHD1 的表达明显高于肥沃对照组。对月经周期的分析表明，EHD1 的表达在中增殖期增加，并在中分泌期和蜕膜期逐渐下降。此外，EHD1 的过表达通过抑制催乳素和胰岛素样生长因子结合蛋白-1 的表达以及细胞骨架的形成，损害了蜕膜化。机制分析表明，EHD1 通过内吞途径调节 LRP5/6 蛋白功能，随后在蜕膜化过程中抑制 Wnt4/β-连环蛋白通路。此外，Wnt4 激动剂改善了受损的蜕膜化过程。
结论
EHD1-Wnt4 通路的调节可能成为改善 RIF 妇女子宫内膜容受性的有前途的治疗策略。
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