Jørgensen Malene Møller, Bæk Rikke, Sloth Jenni K, Sammour Rami, Sharabi-Nov Adi, Vatish Manu, Meiri Hamutal, Sammar Marei
Department of Clinical Immunology, Aalborg University Hospital, Aalborg, Denmark.
Department of Clinical Medicine, Aalborg University, Aalborg, Denmark.
Acta Obstet Gynecol Scand. 2025 Jan;104(1):151-163. doi: 10.1111/aogs.15020. Epub 2024 Nov 28.
Placental-derived extracellular vesicles (EVs) are nano-organelles that facilitate intercellular communication between the feto-placental unit and the mother. We evaluated a novel Multiple Microarray analyzer for identifying surface markers on plasma EVs that predict preterm delivery and preeclampsia compared to term delivery controls.
In this prospective exploratory cohort study pregnant women between 24 and 40 gestational weeks with preterm delivery (n = 16), preeclampsia (n = 19), and matched term delivery controls (n = 15) were recruited from Bnai Zion Medical Center, Haifa, Israel. Plasma samples were tested using a multiple microarray analyzer. Glass slides with 17 antibodies against EV surface receptors - were incubated with raw plasma samples, detected by biotinylated secondary antibodies specific to EVs or placental EVs (PEVs), and labeled with cyanine 5-streptavidin. PBS and whole human IgG served as controls. The fluorescent signal ratio to negative controls was log 2 transformed and analyzed for sensitivity and specificity using the area under the receiver operating characteristics curves (AUROC). Best pair ratios of general EVs/PEVs were used for univariate analysis, and top pairs were combined for multivariate analysis. Results were validated by comparison with EVs purified using standard procedures.
Heatmaps differentiated surface profiles of preeclampsia, preterm delivery, and term delivery receptors on total EVs and PEVs. Similar results were obtained with enriched EVs and EVs from raw plasma. Univariate analyses identified markers predicting preterm delivery and preeclampsia over term delivery controls with AUC >0.6 and sensitivity >50% at 80% specificity. Combining the best markers in a multivariate model, preeclampsia prediction over term delivery had an AUC of 0.89 (95% CI: 0.72-1.0) with 90% sensitivity and 90% specificity, marked by inflammation (TNF RII), relaxation (placenta protein 13 (PP13)), and immune-modulation (LFA1) receptors. Preterm delivery prediction over term delivery had an AUC of 0.97 (0.94-1.0), 84% sensitivity, and 90% specificity, marked by cell adhesion (ICAM), immune suppression, and general EV markers (CD81, CD82, and Alix). Preeclampsia prediction over preterm delivery had an AUC of 0.91 (0.79-0.99) with 80% sensitivity and 90% specificity with markers for complement activation (C1q) and autoimmunity markers.
The new, robust EV Multi-Array analyzer and methodology offer a simple, fast diagnostic tool that reveals novel surface markers for major obstetric syndromes.
胎盘来源的细胞外囊泡（EVs）是一种纳米细胞器，可促进胎儿 - 胎盘单位与母体之间的细胞间通讯。我们评估了一种新型多重微阵列分析仪，用于识别血浆EVs上的表面标志物，以预测早产和先兆子痫，并与足月分娩对照组进行比较。
在这项前瞻性探索性队列研究中，从以色列海法的Bnai Zion医疗中心招募了妊娠24至40周的孕妇，其中早产（n = 16）、先兆子痫（n = 19）以及匹配的足月分娩对照组（n = 15）。使用多重微阵列分析仪检测血浆样本。将带有17种针对EV表面受体抗体的载玻片与原始血浆样本孵育，通过针对EVs或胎盘来源的EVs（PEVs）的生物素化二抗进行检测，并用花青5 - 链霉亲和素标记。磷酸盐缓冲液（PBS）和全人免疫球蛋白G（IgG）作为对照。将荧光信号与阴性对照的比值进行log 2转换，并使用受试者工作特征曲线下面积（AUROC）分析其敏感性和特异性。一般EVs/PEVs的最佳配对比值用于单变量分析，顶级配对组合用于多变量分析。通过与使用标准程序纯化的EVs进行比较来验证结果。
热图区分了先兆子痫、早产和足月分娩受体在总EVs和PEVs上的表面特征。从富集的EVs和原始血浆中的EVs获得了相似的结果。单变量分析确定了在80%特异性时预测早产和先兆子痫高于足月分娩对照组的标志物，其AUC>0.6且敏感性>50%。在多变量模型中结合最佳标志物，预测先兆子痫高于足月分娩的AUC为0.89（95%CI：0.72 - 1.0），敏感性为90%，特异性为90%，标志物为炎症（肿瘤坏死因子受体II（TNF RII））、松弛（胎盘蛋白13（PP13））和免疫调节（淋巴细胞功能相关抗原1（LFA1））受体。预测早产高于足月分娩的AUC为0.97（0.94 - 1.0），敏感性为84%，特异性为90%，标志物为细胞黏附（细胞间黏附分子（ICAM））、免疫抑制和一般EV标志物（分化簇81（CD81）、分化簇82（CD82）和Alix蛋白）。预测先兆子痫高于早产的AUC为0.91（0.79 - 0.99），敏感性为80%，特异性为90%，标志物为补体激活（C1q）和自身免疫标志物。
新型、强大的EV多重阵列分析仪和方法提供了一种简单、快速的诊断工具，可揭示主要产科综合征的新型表面标志物。
