Cai Lei, Lv Mingwei, Wei Jianbo, Liu Chang, Li Yuehan, Liao Zhiqi, Li Tianhui, Zhang Hanwang, Xi Ling, Sui Cong
Reproductive Medicine Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, People's Republic of China.
Department of Gynecological Oncology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People's Republic of China.
Int J Nanomedicine. 2025 May 1;20:5661-5679. doi: 10.2147/IJN.S508491. eCollection 2025.
Recurrent implantation failure (RIF) presents a crucial obstacle to in vitro fertilization success. Previous research has shown that small extracellular vesicles (EVs) from endometrial RIF patients hinder embryo development, yet the underlying mechanism and potential solutions remain largely unexplored. In this study, we aimed to investigate the effectiveness of miR-218-5p as a molecular factor in RIF-EVs. Our findings revealed that miR-218-5p disrupted mouse embryo development, and this effect could be reversed by engineered extracellular vesicles (E-EVs) containing anti-miR-218-5p.
The percentage of blastocyst development and hatching rates, embryo morphology, and the total cell number were measured. RNA-sequencing was used to analyze transcriptional changes in embryos post miR-218-5p agomir treatment. The abnormal segregation genes of trophectoderm (TE) and inner cell mass (ICM) were visualized via qRT-PCR and immunofluorescence staining. The E-EVs were using the EVs derived from Human Umbilical Cord Mesenchymal Stem Cells (HUMSCs). Characteristics of the EVs were measured using Western blotting, nanoparticle tracking analysis, and transmission electron microscopy. EVs internalization was visualized using BODIPY TR ceramide staining.
Mouse embryos were arrested at the morula stage and demonstrated reduced blastocyst and hatching rates following miR-218-5p agomir treatment ( < 0.001). Essential transcription factors for TE and ICM, such as Cdx2, Yap1, Sox2, Nanog, Tead4, were reduced at the mRNA level in the miR-218-5p treated morula. This was accompanied by decreased Cdx2 protein levels at the 8-16-cell stage ( < 0.001) and disruption of co-localization of Yap1 and Cdx2. The blastocyte rate was increased by anti-miR-218-5p-encapsulated E-EVs compared with miR-218-5p group ( < 0.001).
This study offers valuable insights into the potential role of miR-218-5p in RIF and presents. The utilization of engineered vesicles containing anti-miR-218-5p may present a promising avenue for patients facing challenges with RIF.
反复种植失败（RIF）是体外受精成功的一个关键障碍。先前的研究表明，来自子宫内膜RIF患者的小细胞外囊泡（EVs）会阻碍胚胎发育，但其潜在机制和潜在解决方案在很大程度上仍未得到探索。在本研究中，我们旨在研究miR-218-5p作为RIF-EVs中的一种分子因子的有效性。我们的研究结果显示，miR-218-5p会破坏小鼠胚胎发育，而这种作用可以被含有抗miR-218-5p的工程化细胞外囊泡（E-EVs）逆转。
测量囊胚发育百分比、孵化率、胚胎形态和总细胞数。使用RNA测序分析miR-218-5p类似物处理后胚胎的转录变化。通过qRT-PCR和免疫荧光染色观察滋养外胚层（TE）和内细胞团（ICM）的异常分离基因。E-EVs使用源自人脐带间充质干细胞（HUMSCs）的EVs。使用蛋白质免疫印迹法、纳米颗粒跟踪分析和透射电子显微镜测量EVs的特征。使用BODIPY TR神经酰胺染色观察EVs的内化。
miR-218-5p类似物处理后，小鼠胚胎在桑椹胚阶段停滞，囊胚和孵化率降低（<0.001）。在经miR-218-5p处理的桑椹胚中，TE和ICM的关键转录因子，如Cdx2、Yap1、Sox2、Nanog、Tead4，在mRNA水平上降低。这伴随着8-16细胞阶段Cdx2蛋白水平的降低（<0.001）以及Yap1和Cdx2共定位的破坏。与miR-218-5p组相比，封装有抗miR-218-5p的E-EVs提高了囊胚率（<0.001）。
本研究为miR-218-5p在RIF中的潜在作用提供了有价值的见解。对于面临RIF挑战的患者，使用含有抗miR-218-5p的工程化囊泡可能是一条有前景的途径。
