Delcorte Ophélie, Craps Julie, Mahibullah Siam, Spourquet Catherine, D'Auria Ludovic, Van Der Smissen Patrick, Dessy Chantal, Marbaix Etienne, Mourad Michel, Pierreux Christophe E
CELL Unit, de Duve Institute, Université Catholique de Louvain, Brussels, Belgium.
FATH & MORF Unit, IREC, Université Catholique de Louvain, Brussels, Belgium.
Endocr Relat Cancer. 2022 May 16;29(7):389-401. doi: 10.1530/ERC-21-0343.
Differential diagnosis of thyroid cancer and benign nodules is still one of the most challenging issues in the field of endocrinology. To overcome overdiagnosis of papillary thyroid carcinomas (PTC) and the consecutive overtreatment of multinodular diseases, the search for easily accessible, sensitive and accurate biomarkers is critical. Several micro-RNAs (miRNAs) freely circulating in peripheral blood or enclosed in extracellular vesicles (EVs) have been proposed as potential biomarkers from non-invasive liquid biopsies. However, protocols are rarely comparable and conflicting data exist in the literature. In this work, we aimed to assess the diagnostic value of six micro-RNAs by comparing their expression in thyroid tissue to their abundance in bulk plasma and in plasma-EVs, before and after thyroid surgery. Plasma-EVs were isolated using a sequential density- and size-based fractionation, followed by in-depth characterization, confirming EV purity. Micro-RNA levels were measured by RT-qPCR in thyroid tissue, plasma and plasma-EVs. Among the six candidates, only miR-146b-5p and miR-21a-5p displayed a significant differential abundance in purified plasma-derived EVs from patients with PTC and benign disease. However, no difference could be demonstrated in bulk plasma through our cohort of patients. Overall, our work supports the use of a well-defined protocol of plasma-EV miRNAs purification for biomarker discovery, rather than the use of freely circulating miRNAs in bulk plasma. Our work also demonstrates that standardized pre-analytical and analytical procedures as well as optimized EV-miRNAs detection methods are essential.
甲状腺癌与良性结节的鉴别诊断仍是内分泌领域最具挑战性的问题之一。为克服甲状腺乳头状癌（PTC）的过度诊断以及多结节疾病的后续过度治疗，寻找易于获取、敏感且准确的生物标志物至关重要。外周血中自由循环或包裹在细胞外囊泡（EVs）中的几种微小RNA（miRNAs）已被提议作为非侵入性液体活检的潜在生物标志物。然而，相关方案很少具有可比性，且文献中存在相互矛盾的数据。在这项研究中，我们旨在通过比较甲状腺手术前后甲状腺组织中六种微小RNA的表达与其在血浆总体积和血浆来源的细胞外囊泡中的丰度，来评估它们的诊断价值。使用基于密度和大小的连续分级分离法分离血浆来源的细胞外囊泡，随后进行深入表征，以确认细胞外囊泡的纯度。通过逆转录定量聚合酶链反应（RT-qPCR）测量甲状腺组织、血浆和血浆来源的细胞外囊泡中的微小RNA水平。在这六种候选物中，只有miR-146b-5p和miR-21a-5p在来自PTC患者和良性疾病患者的纯化血浆来源的细胞外囊泡中显示出显著的差异丰度。然而，通过我们的患者队列在血浆总体积中未发现差异。总体而言，我们的研究支持使用明确的血浆来源的细胞外囊泡微小RNA纯化方案来发现生物标志物，而不是使用血浆总体积中自由循环的微小RNA。我们的研究还表明，标准化的分析前和分析程序以及优化的细胞外囊泡微小RNA检测方法至关重要。
