Frisk Nanna Lond Skov, Jørgensen Malene Møller, Bæk Rikke, Atic Amila Iriskic, Brodersen Thorsten Rejkjær, Ostrowski Sisse Rye, Larsen Margit Hørup, Posselt Dorthe, Høgdall Estrid, Høgdall Claus, Pedersen Ole Birger Vesterager, Dalgaard Louise Torp
Department of Science and Environment, Roskilde University, Roskilde, Denmark.
Department of Clinical Immunology, Zealand University Hospital, Køge, Denmark.
PLoS One. 2025 May 15;20(5):e0323529. doi: 10.1371/journal.pone.0323529. eCollection 2025.
Ovarian cancer (OC) is the leading cause of gynecological cancer deaths. Current biomarkers of OC are not specific or sensitive enough. Extracellular vesicles (EVs), EV surface proteins and their cargo microRNA (miRNA) show potential as biomarkers. This study aimed to characterize the ability of EVs to identify early OC-biomarkers among blood donors six months before their diagnosis.
Study groups of OC patients, benign tumor patients (B), healthy blood donors (Control), and blood donors with incident OC diagnosis within six months of the last blood draw (Pre-diagnostic; PD) were established. Small EVs were enriched from plasma using ultracentrifugation. EVs were characterized by Dynamic Light Scattering (DLS), EV Array, NanoFlow Cytometry, Nanoparticle Tracking Analysis, and Western blots. RNA from EVs was isolated. A discovery study was performed on OC and B patients using the TaqMan Array Human MicroRNA A card. A validation study of 9 specific miRNAs was performed using RT-qPCR.
With DLS, it was identified that the OC patients' EVs were more heterogeneous in size compared to the other groups. Western blot identified CD63 and TSG101 in the EV enrichments. EV Array assessed 22 known protein biomarkers. TaqMan MicroRNA Array cards indicated a differential miRNA abundance between OC and B; however, technical replication and validation could not validate this pattern.
This study has analyzed EVs in OC, B, Control, and PD women. More extensive investigations of EV CD9, CD151, and CD81 in conjunction with other risk factors and well-known biomarkers like CA125 or HE4 should be the main objectives of future research.
卵巢癌（OC）是妇科癌症死亡的主要原因。目前的OC生物标志物特异性和敏感性不足。细胞外囊泡（EVs）、EV表面蛋白及其携带的微小RNA（miRNA）显示出作为生物标志物的潜力。本研究旨在表征EVs在OC患者诊断前六个月的献血者中识别早期OC生物标志物的能力。
建立OC患者、良性肿瘤患者（B）、健康献血者（对照）以及在最后一次献血后六个月内被诊断为OC的献血者（诊断前；PD）的研究组。使用超速离心从血浆中富集小EVs。通过动态光散射（DLS）、EV阵列、纳米流式细胞术、纳米颗粒跟踪分析和蛋白质印迹对EVs进行表征。分离EVs中的RNA。使用TaqMan Array Human MicroRNA A卡对OC和B患者进行探索性研究。使用RT-qPCR对9种特定miRNA进行验证研究。
通过DLS发现，与其他组相比，OC患者的EVs在大小上更具异质性。蛋白质印迹在EV富集物中鉴定出CD63和TSG101。EV阵列评估了22种已知的蛋白质生物标志物。TaqMan MicroRNA阵列卡显示OC和B之间miRNA丰度存在差异；然而，技术重复和验证未能证实这种模式。
本研究分析了OC、B、对照和PD女性中的EVs。未来研究的主要目标应该是更广泛地研究EV CD9、CD151和CD81，并结合其他风险因素以及CA125或HE4等知名生物标志物。
