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

Ander Arbelaiz, Mikel Azkargorta, Marcin Krawczyk, Alvaro Santos-Laso, Ainhoa Lapitz, Maria J. Perugorria, Oihane Erice, Esperanza Gonzalez, Raul Jimenez-Agüero, Adelaida Lacasta, Cesar Ibarra, Alberto Sanchez-Campos, Juan P. Jimeno, Frank Lammert, Piotr Milkiewicz, Marco Marzioni, Rocio I.R. Macias, Jose J.G. Marin, Tushar Patel, Gregory J. GoresIbon Martinez, Félix Elortza, Juan M. Falcon-Perez, Luis Bujanda, Jesus M. Banales

Research output: Contribution to journalArticlepeer-review

102 Scopus citations

Abstract

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. (Hepatology 2017;66:1125-1143).

Original languageEnglish (US)
Pages (from-to)1125-1143
Number of pages19
JournalHepatology
Volume66
Issue number4
DOIs
StatePublished - Oct 2017

ASJC Scopus subject areas

  • Hepatology

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