Hu Yu-Qin, Zhang Xin-Xian, Zhao Ting-Ting, Wu Xiao-Hua
Hebei Medical University, 361 East Zhongshan Road, Shijiazhuang, Hebei, China.
Reproductive Medicine Center, The Fourth Hospital of Shijiazhuang, Hebei Medical University, Hebei, China.
Arch Gynecol Obstet. 2025 Jun 13. doi: 10.1007/s00404-025-08074-8.
Recurrent implantation failure (RIF) affects about 10% of infertility patients and may involve mid-luteal phase endometrial natural killer (NK) cells. The pathogenesis of NK cells in RIF remains unclear.
Our study integrated proteomics data from endometrial tissues of six RIF patients and six controls, with single-cell sequencing insights.
Our proteomics analysis identified 1366 differentially expressed proteins (DEPs) between RIF and control groups, highlighting alterations in cellular processes, such as cytoplasmic translation and mRNA processing. Functional enrichment analysis revealed significant associations with pathways involved in amyotrophic lateral sclerosis and proteasomes. The DEPs were transformed into differentially expressed genes1 (DEGs1) by ID-transformation. 33 candidate genes were detected when ID-transformed 1210 DEGs1 were intersected with 752 DEGs2 from NK cells. After that, the proteomics sequencing data validation showed that the expression of AMPD3, H6PD, and PAK2 was consistent and significantly different from the GSE111974 dataset and classified as crucial genes. In addition, analysis of single-cell sequencing data annotated fibroblast-like stromal cells, NK cells, T cells, and endothelial cells, and these three essential genes showed that they were expressed in NK cells. Crossing the signaling pathways of key genes with those enriched for DEPs yielded the 'Escherichia coli infection' possibly associated with RIF. Finally, the transcription factor HR had a strong regulatory effect on the PAK2.
Finally, identifying three key genes (AMPD3, H6PD and PAK2) associated with RIF and the regulatory solid roles of HR and PAK2 provided a basis for understanding the molecular mechanism of RIF. Our findings may pave the way for developing targeted therapies to improve pregnancy outcomes in patients with RIF.
反复种植失败（RIF）影响约10%的不孕患者，可能与黄体中期子宫内膜自然杀伤（NK）细胞有关。RIF中NK细胞的发病机制尚不清楚。
我们的研究整合了6例RIF患者和6例对照者子宫内膜组织的蛋白质组学数据，并结合单细胞测序见解。
我们的蛋白质组学分析确定了RIF组和对照组之间1366种差异表达蛋白（DEP），突出了细胞过程中的改变，如细胞质翻译和mRNA加工。功能富集分析显示与肌萎缩侧索硬化和蛋白酶体相关的途径有显著关联。通过ID转换将DEP转化为差异表达基因1（DEG1）。当将1210个DEG1进行ID转换后与来自NK细胞的752个DEG2相交时，检测到33个候选基因。之后，蛋白质组学测序数据验证表明，AMPD3、H6PD和PAK2的表达一致，且与GSE111974数据集有显著差异，并被归类为关键基因。此外，单细胞测序数据分析注释了成纤维细胞样基质细胞、NK细胞、T细胞和内皮细胞，这三个关键基因在NK细胞中表达。关键基因的信号通路与DEP富集的信号通路交叉产生了可能与RIF相关的“大肠杆菌感染”。最后，转录因子HR对PAK2有很强的调控作用。
最后，确定了与RIF相关的三个关键基因（AMPD3、H6PD和PAK2）以及HR和PAK2的调控作用，为理解RIF的分子机制提供了依据。我们的发现可能为开发靶向治疗方法以改善RIF患者的妊娠结局铺平道路。
