A specific technique of immunolabelling of urinary small extracellular vesicle biomarkers for the diagnostic of renal cancer.

Microscopy (Oxford, England) Pub Date : 2023-10-09 DOI:10.1093/jmicro/dfad007

Nora Mallouk, Sisareuth Tan, Claire Scepaniack, Arnauld Garcin, Camille Pradat, Claude Lambert, Guorong Li

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Abstract

Small extracellular vesicles (EVs) are characterized by the membrane expression of CD63, CD81 and CD9 tetraspanins. Their size is inferior to 200 nm. They share the same characteristics as the native cells and are found in human fluids. Specific membrane protein biomarkers expressed on small EV are useful for the diagnosis of tumoural pathologies. Clear cell renal cell carcinoma (CCRCC) is diagnosed by imaging examinations and/or tissue biopsy. Carbonic anhydrase IX (CAIX) is a powerful biomarker of CCRCC. The detection of CAIX on small EV from the urine of patients could constitute a liquid biopsy for CCRCC. We have set up a specific protocol for the preparation, the immunostaining characterization and the transmission electron microscopy observation of small EVs isolated from the urine of CCRCC patients. The background labelling was significantly reduced. We successfully detected biomarkers on urinary small EVs from CCRCC patients. This technique could be extended with antibodies directed against other EV biomarkers for the detection and the monitoring of cancer diseases.

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尿小细胞外囊泡生物标志物免疫标记技术在癌症诊断中的应用。

细胞外小泡（EVs）的特征是CD63、CD81和CD9四跨膜蛋白的膜表达。它们的尺寸低于200 nm。它们与天然细胞具有相同的特征，并且存在于人体体液中。在小EV上表达的特异性膜蛋白生物标志物可用于诊断肿瘤病理。肾透明细胞癌是通过影像学检查和/或组织活检来诊断的。碳酸酐酶IX（CAIX）是CCRC的一个强大的生物标志物。从患者尿液中检测小EV上的CAIX可以构成CCRCC的液体活检。我们已经为从CCRCC患者尿液中分离的小EVs的制备、免疫染色表征和透射电子显微镜观察建立了一个特定的方案。背景标记显著减少。我们成功地检测到CCRCC患者尿中小型EVs的生物标志物。这项技术可以扩展到针对其他EV生物标志物的抗体，用于检测和监测癌症疾病。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Microscopy (Oxford, England)

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