Table_4_Exometabolomic Analysis of Decidualizing Human Endometrial Stromal and Perivascular Cells.XLSX

Differentiation of endometrial fibroblasts into specialized decidual cells controls embryo implantation and transforms the cycling endometrium into a semi-permanent, immune-protective matrix that accommodates the placenta throughout pregnancy. This process starts during the midluteal phase of the menstrual cycle with decidual transformation of perivascular cells (PVC) surrounding the terminal spiral arterioles and endometrial stromal cells (EnSC) underlying the luminal epithelium. Decidualization involves extensive cellular reprogramming and acquisition of a secretory phenotype, essential for coordinated placental trophoblast invasion. Secreted metabolites are an emerging class of signaling molecules, collectively known as the exometabolome. Here, we used liquid chromatography-mass spectrometry to characterize and analyze time-resolved changes in metabolite secretion (exometabolome) of primary PVC and EnSC decidualized over 8 days. PVC were isolated using positive selection of the cell surface marker SUSD2. We identified 79 annotated metabolites differentially secreted upon decidualization, including prostaglandin, sphingolipid, and hyaluronic acid metabolites. Secreted metabolites encompassed 21 metabolic pathways, most prominently glycerolipid and pyrimidine metabolism. Although temporal exometabolome changes were comparable between decidualizing PVC and EnSC, 32 metabolites were differentially secreted across the decidualization time-course. Further, targeted metabolomics demonstrated significant differences in secretion of purine pathway metabolites between decidualized PVC and EnSC. Taken together, our findings indicate that the metabolic footprints generated by different decidual subpopulations encode spatiotemporal information that may be important for optimal embryo implantation.

子宫内膜成纤维细胞向特化蜕膜细胞（decidual cell）的分化，可调控胚胎植入过程，并将周期性子宫内膜转化为可在整个妊娠期间容纳胎盘的半永久性免疫保护基质。该过程始于月经周期的黄体中期，表现为末端螺旋小动脉周围的血管周围细胞（perivascular cell, PVC）以及腔上皮下的子宫内膜基质细胞（endometrial stromal cell, EnSC）发生蜕膜化转变。蜕膜化涉及广泛的细胞重编程与分泌表型的获得，这对于协调胎盘滋养层侵袭至关重要。分泌代谢物是一类新兴的信号分子，统称为外代谢组（exometabolome）。本研究采用液相色谱-质谱联用技术，对原代PVC与EnSC在8天蜕膜化培养过程中分泌代谢物（外代谢组）的时间序列变化进行表征与分析。研究通过细胞表面标志物SUSD2的阳性分选法分离得到PVC。本研究共鉴定出79种在蜕膜化过程中差异分泌的注释代谢物，涵盖前列腺素、鞘脂及透明质酸代谢物等类别。这些分泌代谢物涉及21条代谢通路，其中以甘油脂代谢和嘧啶代谢最为突出。尽管正在进行蜕膜化的PVC与EnSC的时间序列外代谢组变化趋势相近，但在整个蜕膜化时间进程中，共有32种代谢物的分泌水平存在显著差异。此外，靶向代谢组学分析显示，完全蜕膜化的PVC与EnSC的嘌呤通路代谢物分泌水平存在显著差异。综上，本研究结果表明，不同蜕膜亚群产生的代谢印迹携带着时空信息，这可能对实现最优胚胎植入具有重要意义。