Serum Extracellular Vesicles Contain Protein Biomarkers for Primary Sclerosing Cholangitis and Cholangiocarcinoma
Serum Extracellular Vesicles Contain Protein Biomarkers for Primary Sclerosing Cholangitis and Cholangiocarcinoma
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DOI：
10.1002/hep.29291
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发表时间：
2017-10-01
期刊：
影响因子：
13.5
通讯作者：
Banales, Jesus M.
中科院分区：
文献类型：
--
作者：
Arbelaiz, Ander;Azkargorta, Mikel;Banales, Jesus M.
胆管癌（CCA）包括一组异质性的胆管癌，预后不良。几种情况，如原发性硬化性胆管炎（PSC），是危险因素。肝内CCA和肝细胞癌（HCC）之间的非侵入性鉴别诊断有时很困难。PSC、CCA和HCC的准确非侵入性生物标志物不可用。在寻找新的生物标志物中，从CCA（n = 43）、PSC（n = 30）或HCC（n = 29）患者和健康个体（对照，n = 32）分离血清细胞外囊泡（EV）;并表征其蛋白质含量。通过使用纳米粒子跟踪分析，发现血清EV浓度在HCC中高于所有其他组。圆形形态（通过透射电子显微镜）、大小（通过纳米颗粒跟踪分析，直径类似于180 nm）和标记物（通过免疫印迹，分化簇9、63和81）表明大多数血清EV是外来体。蛋白质组图谱（质谱法）显示多个组间差异表达的蛋白质。这些蛋白质中的几种显示出高诊断值，其中CCA与对照相比的接收器操作特征曲线下的最大面积为0.878，CCA I-II期与对照相比的最大面积为0.905，PSC与对照相比的最大面积为0.789，非肝硬化PSC与对照相比的最大面积为0.806，CCA与PSC相比的最大面积为0.796，CCA I-II期与PSC相比的最大面积为0.956，0.904，HCC与对照组，0.894，肝内CCA与HCC。蛋白质组学分析的EV来自CCA人类细胞在体外显示较高丰度的致癌蛋白相比，EV释放的正常人胆管细胞。将CCA人细胞原位移植到免疫缺陷小鼠的肝脏中，导致EV释放到血清中，其中含有一些类似的人致癌蛋白。结论：在CCA、PSC和HCC患者的血清EV中发现的蛋白质组特征显示出作为诊断工具的潜在有用性。
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 (similar to 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.