Kothandan Vinoth K, Ouyang Yingshi, Sadovsky Elena, Komsky-Elbaz Alisa, Powell Juliana S, Xia Jianping, Huang Tony J, Sadovsky Yoel
Magee-Womens Research Institute, Department of Obstetrics, Gynecology and Reproductive Sciences, University of Pittsburgh, Pittsburgh, PA, USA.
Magee-Womens Research Institute, Department of Obstetrics, Gynecology and Reproductive Sciences, University of Pittsburgh, Pittsburgh, PA, USA; Department of Entomology, The Hebrew University of Jerusalem, Rehovot, Israel.
Placenta. 2025 Jun 13;166:96-102. doi: 10.1016/j.placenta.2024.12.014. Epub 2024 Dec 28.
MicroRNAs (miRNAs), packaged within extracellular vesicles (EVs), have been used to interrogate the pathogenesis of preeclampsia and to identify its biomarkers. We have previously shown that miRNA species were differentially expressed in small plasma EVs from women with preeclampsia vs healthy controls. We sought to assess the use of rapid technologies for isolation of plasma and urine EVs from parturients with preeclampsia and determine differences in the expression of selected EV miRNA species.
We collected blood and urine samples before delivery from parturients with severe preeclampsia vs healthy controls and used size exclusion chromatography (SEC) as an acceptable standard for rapid isolation of plasma EVs. We also isolated urine and plasma EVs using ExoDisc, a rapid nanofiltration technology for EV isolation. All samples were examined using a nanoparticle tracking analyzer, immunoblotting, and RT-qPCR for selected miRNA levels.
Whereas the concentration of EVs was higher in the urine from preeclampsia compared to controls, we observed the opposite change in plasma EVs, with no difference in EV size. Comparing the two patient groups for miRNA levels in EVs isolated by ExoDisc or SEC, we found that EV miR-93-5p was upregulated in the plasma and urine of parturients with preeclampsia vs healthy controls. Notably, miR-31-5p was upregulated in SEC- or ExoDisc-isolated plasma EVs, and miR-92-3p was upregulated in or ExoDisc-isolated plasma or urine EVs of parturients with preeclampsia.
Technologies for rapid analysis of plasma and urine EVs and their miRNA cargo provide complementary information that might be useful for deciphering pathways leading to preeclampsia and biomarkers for this disease.
包裹在细胞外囊泡（EVs）中的微小RNA（miRNAs）已被用于探究子痫前期的发病机制并识别其生物标志物。我们之前已经表明，子痫前期患者与健康对照者的小血浆EVs中miRNA种类存在差异表达。我们试图评估使用快速技术从子痫前期产妇中分离血浆和尿液EVs，并确定所选EV miRNA种类表达的差异。
我们在分娩前收集了重度子痫前期产妇与健康对照者的血液和尿液样本，并使用尺寸排阻色谱法（SEC）作为快速分离血浆EVs的可接受标准。我们还使用ExoDisc（一种用于EV分离的快速纳米过滤技术）分离尿液和血浆EVs。所有样本均使用纳米颗粒跟踪分析仪、免疫印迹法和RT-qPCR检测所选miRNA水平。
与对照组相比，子痫前期患者尿液中EVs的浓度更高，而我们观察到血浆EVs出现相反的变化，EV大小无差异。比较通过ExoDisc或SEC分离的EVs中两组患者的miRNA水平，我们发现子痫前期产妇与健康对照者相比，血浆和尿液中的EV miR-93-5p上调。值得注意的是，miR-31-5p在SEC或ExoDisc分离的血浆EVs中上调，miR-92-3p在子痫前期产妇的ExoDisc分离的血浆或尿液EVs中上调。
快速分析血浆和尿液EVs及其miRNA含量的技术提供了互补信息，这可能有助于解读导致子痫前期的途径以及该疾病的生物标志物。
