Serum extracellular vesicles contain protein biomarkers for primary sclerosing cholangitis and cholangiocarcinoma

纳米粒子跟踪分析 原发性硬化性胆管炎 肝内胆管癌 肝细胞癌 外体 细胞外小泡 蛋白质组 微泡 胞外囊泡 接收机工作特性 生物 曲线下面积 内科学 胃肠病学 病理 癌症研究 化学 医学 小RNA 生物信息学 基因 生物化学 疾病 细胞生物学
作者
Ander Arbelaiz,Mikel Azkargorta,Marcin Krawczyk,Álvaro Santos‐Laso,Ainhoa Lapitz,María J. Perugorria,Oihane Erice,Esperanza González,Raúl Jiménez-Agüero,Adelaida La Casta,Cesar Ibarra,Alberto Sanchez‐Campos,J.P. Jimeno,Frank Lammert,Piotr Milkiewicz,Marco Marzioni,Rocı́o I.R. Macı́as,José J.G. Marı́n,Tushar Patel,Gregory J. Gores
出处
期刊:Hepatology [Lippincott Williams & Wilkins]
卷期号:66 (4): 1125-1143 被引量:253
标识
DOI:10.1002/hep.29291
摘要

Cholangiocarcinoma (CCA) includes a heterogeneous group of biliary cancers with poor prognosis. Several conditions, such as primary sclerosing cholangitis (PSC), are risk factors. Noninvasive differential diagnosis between intrahepatic CCA and hepatocellular carcinoma (HCC) is sometimes difficult. Accurate noninvasive biomarkers for PSC, CCA, and HCC are not available. In the search for novel biomarkers, serum extracellular vesicles (EV) were isolated from CCA (n = 43), PSC (n = 30), or HCC (n = 29) patients and healthy individuals (control, n = 32); and their protein content was characterized. By using nanoparticle tracking analysis, serum EV concentration was found to be higher in HCC than in all the other groups. Round morphology (by transmission electron microscopy), size (∼180 nm diameter by nanoparticle tracking analysis), and markers (clusters of differentiation 9, 63, and 81 by immunoblot) indicated that most serum EV were exosomes. Proteome profiles (by mass spectrometry) revealed multiple differentially expressed proteins among groups. Several of these proteins showed high diagnostic values with maximum area under the receiver operating characteristic curve of 0.878 for CCA versus control, 0.905 for CCA stage I‐II versus control, 0.789 for PSC versus control, 0.806 for noncirhottic PSC versus control, 0.796 for CCA versus PSC, 0.956 for CCA stage I‐II versus PSC, 0.904 for HCC versus control, and 0.894 for intrahepatic CCA versus HCC. Proteomic analysis of EV derived from CCA human cells in vitro revealed higher abundance of oncogenic proteins compared to EV released by normal human cholangiocytes. Orthotopic implant of CCA human cells in the liver of immunodeficient mice resulted in the release to serum of EV containing some similar human oncogenic proteins. Conclusion : Proteomic signatures found in serum EV of CCA, PSC, and HCC patients show potential usefulness as diagnostic tools. (H epatology 2017;66:1125‐1143).
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