Data_Sheet_3_Exometabolomic Analysis of Decidualizing Human Endometrial Stromal and Perivascular Cells.PDF

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分化为特化蜕膜细胞的过程，可调控胚胎植入，并将周期性增殖的子宫内膜转化为半永久性、具有免疫保护作用的基质，以在整个妊娠过程中容纳胎盘。该过程始于月经周期的黄体中期，表现为终端螺旋动脉周围的血管周细胞（perivascular cells, PVC）以及腔上皮下方的子宫内膜基质细胞（endometrial stromal cells, EnSC）发生蜕膜化转化。蜕膜化过程涉及广泛的细胞重编程与分泌表型的获得，这是协同调控胎盘滋养层侵袭的关键要素。分泌型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（liquid chromatography-mass spectrometry），对原代PVC与EnSC在8天蜕膜化过程中的代谢物分泌（外代谢组）的时间动态变化进行表征与分析。研究人员通过细胞表面标志物SUSD2的阳性分选分离得到PVC。本研究共鉴定出79种在蜕膜化过程中差异分泌的注释型代谢物，包括前列腺素、鞘脂类及透明质酸代谢物。分泌型代谢物涵盖21条代谢通路，其中以甘油脂代谢与嘧啶代谢最为显著。尽管蜕膜化的PVC与EnSC的外代谢组时间动态变化趋势相近，但在整个蜕膜化时间进程中，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（targeted metabolomics）分析显示，蜕膜化的PVC与EnSC之间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特征携带有时空信息，这可能对优化胚胎植入过程具有重要意义。