标题： 胎盘滋养层细胞的单细胞测序以及BeWo b30细胞中的化学缺氧揭示了EBI3、COL17A1、miR-27a-5p和miR-193b-5p作为缺氧反应标志物 显示英文标题

标题： Single-cell sequencing of trophoblasts in preeclampsia and chemical hypoxia in BeWo b30 cells reveals EBI3, COL17A1, miR-27a-5p, and miR-193b-5p as hypoxia-response markers

摘要： 背景。 子痫前期（PE）影响2-8%的妊娠，其特征是胎盘缺氧和HIF通路激活，尤其是在早发性PE（eoPE）中。 将患者组织分析与实验模型结合可能揭示滋养层细胞缺氧的共同分子标志物。 方法。 我们分析了10例eoPE、7例晚发性PE（loPE）及其对应对照胎盘的单细胞RNA测序数据，鉴定了绒毛合体滋养细胞（VCT）、合胞体滋养细胞（SCT）和绒毛外滋养细胞（EVT）亚群。 BeWo b30细胞用CoCl2（300 $\mu$M）或氧喹啉衍生物（OD，5 $\mu$M）处理24小时以诱导缺氧。 RNA和小RNA测序量化了mRNA和微RNA的变化。 PROGENy推断了通路活性。 结果。 单细胞RNA测序显示eoPE中缺氧通路激活最高，其中EVT在滋养层细胞群体中表现出最大活性。 eoPE中所有滋养层类型均上调了9个基因：EBI3、CST6、FN1、RFK、COL17A1、LDHA、PKP2、RPS4Y1和RPS26。 体外实验中，OD比CoCl2诱导了更特异的缺氧反应，分别有1,284和3,032个差异表达基因。 关键的是，EBI3、FN1和COL17A1在胎盘组织和OD处理的细胞中显示出一致的上调，而CoCl2处理则产生了相反的表达模式。 微RNA分析确定hsa-miR-27a-5p和hsa-miR-193b-5p在两种实验条件下均持续升高，并且之前在PE胎盘囊泡中被报道过。 我们还鉴定了hsa-miR-9-5p和hsa-miR-92b-3p的异构体作为滋养层缺氧相关。 结论。 EBI3、COL17A1、hsa-miR-27a-5p和hsa-miR-193b-5p作为PE中滋养层缺氧的标志物出现。 氧喹啉衍生物比CoCl2提供了一个更生理相关的体外缺氧模型。 这种综合方法推进了对PE病理生理学的理解，并提出了候选治疗靶点。 显示英文摘要

摘要： Background. Preeclampsia (PE) complicates 2-8% of pregnancies and is marked by placental hypoxia and HIF-pathway activation, especially in early-onset PE (eoPE). Integrating patient tissue analyses with experimental models may reveal common molecular markers of trophoblast hypoxia. Methods. We analyzed scRNA-seq data from 10 eoPE, 7 late-onset PE (loPE), and corresponding control placentas, identifying villous cytotrophoblast (VCT), syncytiotrophoblast (SCT), and extravillous trophoblast (EVT) subpopulations. BeWo b30 cells were treated for 24 h with CoCl2 (300 $\mu$M) or an oxyquinoline derivative (OD, 5 $\mu$M) to induce hypoxia. RNA and small RNA sequencing quantified mRNA and microRNA changes. PROGENy inferred pathway activities. Results. ScRNA-seq revealed highest hypoxia pathway activation in eoPE, with EVT showing maximum activity among trophoblast populations. Nine genes were upregulated across all trophoblast types in eoPE: EBI3, CST6, FN1, RFK, COL17A1, LDHA, PKP2, RPS4Y1, and RPS26. In vitro, OD induced more specific hypoxia responses than CoCl2, with 1,284 versus 3,032 differentially expressed genes respectively. Critically, EBI3, FN1, and COL17A1 showed concordant upregulation in both placental tissue and OD-treated cells, while CoCl2 treatment produced opposite expression patterns. MicroRNA analysis identified hsa-miR-27a-5p and hsa-miR-193b-5p as consistently elevated in both experimental conditions and previously reported in PE placental vesicles. We also identified isoforms of hsa-miR-9-5p and hsa-miR-92b-3p as hypoxia-associated in trophoblast. Conclusions. EBI3, COL17A1, hsa-miR-27a-5p, and hsa-miR-193b-5p emerge as trophoblast hypoxia markers in PE. Oxyquinoline derivatives offer a more physiologically relevant in vitro hypoxia model than CoCl2. This integrated approach advances understanding of PE pathophysiology and suggests candidate therapeutic targets.