由于缺乏可靠的生物标志物，区分良性和恶性胰胆疾病具有挑战性。循环细胞外囊泡（EV）已成为细胞之间的功能介质。他们的货物，包括 microRNA (miRNA)，越来越多地被认为是潜在生物标志物的重要来源。这项多中心、前瞻性研究旨在建立诊断性血浆 EV 衍生的 miRNA 特征，以区分胰腺导管腺癌 (PDAC) 和良性胰胆疾病。使用尺寸排阻色谱 (SEC) 分离血浆 EV，并使用纳米颗粒跟踪分析、电子显微镜和蛋白质印迹法。 EV-RNA 经过小 RNA 测序，以发现 PDAC 的差异表达标记（n = 10 个良性 vs. 10 个 PDAC）。然后通过 RT-qPCR 在 61 名患者（n = 31 名良性患者与 30 名 PDAC 患者）组成的队列中验证候选 EV-miRNA。使用逻辑回归和最佳阈值（约登指数）开发 EV-miR-200 家族模型来检测癌症。该模型在 95 名患者的独立队列中进行了测试（n = 30 例良性患者、33 例 PDAC 和 32 例胆管癌）。小 RNA 测序和 RT-qPCR 显示，与良性疾病相比，EV-miR-200 家族成员在 PDAC 中显着过表达。 EV-miR-200 家族的联合表达显示 AUC 为 0.823。在一个独立的验证队列中，应用该模型显示诊断 PDAC 的敏感性、特异性和 AUC 分别为 100%、88% 和 0.97。这是第一项验证血浆 EV-miR-200 成员作为临床药物的研究-PDAC 有用的诊断生物标志物。在更大的队列和临床试验中进一步验证至关重要。这些发现还表明了监测反应和/或复发的潜在效用。© 2024。作者。

Distinguishing benign from malignant pancreaticobiliary disease is challenging because of the absence of reliable biomarkers. Circulating extracellular vesicles (EVs) have emerged as functional mediators between cells. Their cargos, including microRNAs (miRNAs), are increasingly acknowledged as an important source of potential biomarkers. This multicentric, prospective study aimed to establish a diagnostic plasma EV-derived miRNA signature to discriminate pancreatic ductal adenocarcinoma (PDAC) from benign pancreaticobiliary disease.Plasma EVs were isolated using size exclusion chromatography (SEC) and characterised using nanoparticle tracking analysis, electron microscopy and Western blotting. EV-RNAs underwent small RNA sequencing to discover differentially expressed markers for PDAC (n = 10 benign vs. 10 PDAC). Candidate EV-miRNAs were then validated in a cohort of 61 patients (n = 31 benign vs. 30 PDAC) by RT-qPCR. Logistic regression and optimal thresholds (Youden Index) were used to develop an EV-miR-200 family model to detect cancer. This model was tested in an independent cohort of 95 patients (n = 30 benign, 33 PDAC, and 32 cholangiocarcinoma).Small RNA sequencing and RT-qPCR showed that EV-miR-200 family members were significantly overexpressed in PDAC vs. benign disease. Combined expression of the EV-miR-200 family showed an AUC of 0.823. In an independent validation cohort, application of this model showed a sensitivity, specificity and AUC of 100%, 88%, and 0.97, respectively, for diagnosing PDAC.This is the first study to validate plasma EV-miR-200 members as a clinically-useful diagnostic biomarker for PDAC. Further validation in larger cohorts and clinical trials is essential. These findings also suggest the potential utility in monitoring response and/or recurrence.© 2024. The Author(s).