Endometrial gland impacts on stromal cell decidualization

Uterine glands and, by inference, their secretions impact uterine receptivity, blastocyst implantation, stromal cell decidualization, and placental development. Changes in gland function across the menstrual cycle are impacted by steroid hormones, estrogen and progesterone, as well as stroma-derived factors. Using an endometrial epithelial organoid (EEO) system, transcriptome and proteome analyses identified distinct responses of the EEO to steroid hormones and prostaglandin E2 (PGE2). Notably, steroid hormones and PGE2 modulated the basolateral secretion of EEO proteins, where cystatin C (CST3) was significantly increased by progesterone and PGE2. CST3 treatment of decidualizing stromal cells significantly decreased the decidualization markers PRL and IGFBP1. The attenuation of stromal cell decidualization via CST3 suggests a role for uterine gland-derived proteins in controlling the extent of decidualization. These findings provide evidence that uterine gland-derived factors directly impact stromal cell decidualization, which has strong implications for better understanding pregnancy establishment and female fertility in humans. Pair-end sequencing was performed on hormone treated EEO derived from three donors. FastQC (v 0.11.9) and MultiQC (v 1.10.1) were used to evaluate the quality of the sequence. Trimmomatic (v 1.10.1) was used to remove adapters and low-quality bases from the reads. Trimmed reads were aligned to the human genome (GRCh38) with STAR aligner (v 2.7.9a) (average unique mapping rate of 89.21% and read length of 200 bp). Gene quantification was calculated by using featureCounts from Subread package (v 2.0.3). Counts extracted from featureCounts were used to perform differential gene analysis in R (version 4.0.2) using DESeq2 (v 1.30.1).

子宫腺体及其分泌物（据此推测）可影响子宫容受性（uterine receptivity）、囊胚着床（blastocyst implantation）、基质细胞蜕膜化（stromal cell decidualization）以及胎盘发育（placental development）。月经周期中腺体功能的变化受类固醇激素（steroid hormones）、雌激素（estrogen）、孕激素（progesterone）以及基质来源因子的调控。本研究利用子宫内膜上皮类器官（endometrial epithelial organoid, EEO）模型，通过转录组与蛋白质组分析发现，EEO对类固醇激素及前列腺素E2（prostaglandin E2, PGE2）存在特异性应答。值得注意的是，类固醇激素与PGE2可调控EEO蛋白的基外侧分泌（basolateral secretion），其中孕激素与PGE2可显著上调胱抑素C（cystatin C, CST3）的表达。对蜕膜化基质细胞施加CST3处理后，蜕膜化标志物催乳素（PRL）与胰岛素样生长因子结合蛋白1（IGFBP1）的水平显著降低。通过CST3实现的基质细胞蜕膜化减弱现象，提示子宫腺体来源的蛋白在调控蜕膜化程度中发挥作用。上述研究结果证实，子宫腺体来源的因子可直接影响基质细胞蜕膜化，这对深入理解人类妊娠建立与女性生育能力具有重要意义。本研究对来自3名供体的激素处理EEO样本进行了双端测序（pair-end sequencing）。使用FastQC（v0.11.9）与MultiQC（v1.10.1）评估序列质量，通过Trimmomatic（v1.10.1）去除测序读段中的接头序列与低质量碱基。将质控后的测序读段通过STAR比对工具（STAR aligner, v2.7.9a）比对至人类基因组（GRCh38），最终平均唯一比对率为89.21%，测序读段平均长度为200 bp。使用Subread软件包（v2.0.3）中的featureCounts进行基因定量。将featureCounts得到的计数矩阵导入R语言（version 4.0.2），利用DESeq2（v1.30.1）进行差异基因分析。