title: Cancer-associated circulating large extracellular vesicles in cholangiocarcinoma and hepatocellular carcinoma.
abstract: Large extracellular vesicles, specifically AnnexinV+EpCAM+CD147+tumour-associated microparticles (taMPs), facilitate the detection of colorectal carcinoma (CRC), non-small cell lung carcinoma (NSCLC) as well as pancreas carcinoma (PaCa). Here we assess the diagnostic value of taMPs for detection and monitoring of hepatocellular carcinoma (HCC) and cholangiocarcinoma (CCA). Specifically, the aim of this study was to differentiate liver taMPs from other cancer taMPs, such as CRC and NSCLC.
  
  
  
  Fluorescence-activated cell scanning (FACS) was applied to detect various taMP populations in patients' sera that were associated with the presence of a tumour (AnnexinV+EpCAM+CD147+taMPs) or could discriminate between cirrhosis (due to HCV or HBV) and liver cancers (AnnexinV+EpCAM+ASGPR1+taMPs). In total 172 patients with liver cancer (HCC or CCA), 54 with cirrhosis and no liver neoplasia, and 202 control subjects were enrolled.
  
  
  
  The results indicate that AnnexinV+EpCAM+CD147+taMPs were elevated in HCC and CCA. Furthermore, AnnexinV+EpCAM+ASGPR1+CD133+taMPs allowed the distinction of liver malignancies (HCC or CCA) and cirrhosis from tumour-free individuals and, more importantly, from patients carrying other non-liver cancers. In addition, AnnexinV+EpCAM+ASGPR1+taMPs were increased in liver cancer-bearing patients compared to patients with cirrhosis that lacked any detectable liver malignancy. The smallest sizes of successfully detected cancers were ranging between 11-15mm. AnnexinV+EpCAM+ASGPR1+taMPs decreased at 7days after curative R0 tumour resection suggesting close correlations with tumour presence. ROC values, sensitivity/specificity scores and positive/negative predictive values (>78%) indicated a potent diagnostic accuracy of AnnexinV+EpCAM+ASGPR1+taMPs.
  
  
  
  These data provide strong evidence that AnnexinV+EpCAM+ASGPR1+taMPs are a novel biomarker of HCC and CCA liquid biopsy that permit a non-invasive assessment of the presence and possible extent of these cancers in patients with advanced liver diseases.
  
  
  
  Microparticles (MPs) are small vesicles that bleb from the membrane of every cell, including cancer cells, and are released to circulate in the bloodstream. Since their surface composition is similar to the surface of their underlying parental cell, MPs from the bloodstream can be isolated and by screening their surface components, the presence of their parental cells can be identified. This way, it was possible to detect and discriminate between patients bearing liver cancer and chronic liver cirrhosis.
authors.affiliation: Department of Medicine II, Saarland University Medical Center, Saarland University, Homburg, Germany.
authors.forename: Henrike
authors.initials: H
authors.lastname: Julich-Haertel
authors.affiliation: Department of Medicine II, Saarland University Medical Center, Saarland University, Homburg, Germany.
authors.forename: Sabine K
authors.initials: SK
authors.lastname: Urban
authors.affiliation: Department of Medicine II, Saarland University Medical Center, Saarland University, Homburg, Germany; Laboratory of Metabolic Liver Diseases, Department of General, Transplant and Liver Surgery, Medical University of Warsaw, Warsaw, Poland.
authors.forename: Marcin
authors.initials: M
authors.lastname: Krawczyk
authors.affiliation: Department of General, Visceral and Thoracic Surgery, German Armed Forces Central Hospital, Koblenz, Germany.
authors.forename: Arnulf
authors.initials: A
authors.lastname: Willms
authors.affiliation: Department Internal Medicine and Cardiology, Medical University of Warsaw, Warsaw, Poland.
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Jankowski
authors.affiliation: Department of General, Transplant and Liver Surgery, Medical University of Warsaw, Warsaw, Poland.
authors.forename: Waldemar
authors.initials: W
authors.lastname: Patkowski
authors.affiliation: Laboratory of Metabolic Liver Diseases, Department of General, Transplant and Liver Surgery, Medical University of Warsaw, Warsaw, Poland.
authors.forename: Beata
authors.initials: B
authors.lastname: Kruk
authors.affiliation: Department of General, Transplant and Liver Surgery, Medical University of Warsaw, Warsaw, Poland.
authors.forename: Maciej
authors.initials: M
authors.lastname: Krasnodębski
authors.affiliation: Department of General, Transplant and Liver Surgery, Medical University of Warsaw, Warsaw, Poland.
authors.forename: Joanna
authors.initials: J
authors.lastname: Ligocka
authors.affiliation: Department of General, Visceral and Thoracic Surgery, German Armed Forces Central Hospital, Koblenz, Germany.
authors.forename: Robert
authors.initials: R
authors.lastname: Schwab
authors.affiliation: Department of General, Visceral and Thoracic Surgery, German Armed Forces Central Hospital, Koblenz, Germany.
authors.forename: Ines
authors.initials: I
authors.lastname: Richardsen
authors.affiliation: Department of General, Visceral and Thoracic Surgery, German Armed Forces Central Hospital, Koblenz, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Schaaf
authors.affiliation: Department of General, Visceral and Thoracic Surgery, German Armed Forces Central Hospital, Koblenz, Germany.
authors.forename: Angelina
authors.initials: A
authors.lastname: Klein
authors.affiliation: Department of Molecular and Cellular Sport Medicine, Institute of Cardiovascular Research and Sport Medicine, German Sport University Cologne, Cologne, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Gehlert
authors.affiliation: Department of Medicine II, Saarland University Medical Center, Saarland University, Homburg, Germany.
authors.forename: Hanna
authors.initials: H
authors.lastname: Sänger
authors.affiliation: Department of Medicine II, Saarland University Medical Center, Saarland University, Homburg, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Casper
authors.affiliation: Department of Liver and Gastrointestinal Diseases, Biodonostia Health Research Institute - Donostia University Hospital, University of the Basque Country (UPV/EHU), CIBERehd, Ikerbasque, San Sebastian, Spain.
authors.forename: Jesus M
authors.initials: JM
authors.lastname: Banales
authors.affiliation: Institute of Translational Immunology and Research Center for Immune Therapy, University Medical Center, Johannes Gutenberg University, Mainz, Germany.
authors.forename: Detlef
authors.initials: D
authors.lastname: Schuppan
authors.affiliation: Liver and Internal Medicine Unit, Department of General, Transplant and Liver Surgery, Medical University of Warsaw, Warsaw, Poland; Translational Medicine Group, Pomeranian Medical University, Szczecin, Poland.
authors.forename: Piotr
authors.initials: P
authors.lastname: Milkiewicz
authors.affiliation: Department of Medicine II, Saarland University Medical Center, Saarland University, Homburg, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Lammert
authors.affiliation: Department of General, Transplant and Liver Surgery, Medical University of Warsaw, Warsaw, Poland.
authors.forename: Marek
authors.initials: M
authors.lastname: Krawczyk
authors.affiliation: Department of Medicine II, Saarland University Medical Center, Saarland University, Homburg, Germany.
authors.forename: Veronika
authors.initials: V
authors.lastname: Lukacs-Kornek
authors.affiliation: Department of Medicine II, Saarland University Medical Center, Saarland University, Homburg, Germany; Department of General, Visceral and Thoracic Surgery, German Armed Forces Central Hospital, Koblenz, Germany. Electronic address: miroslawkornek@web.de.
authors.forename: Miroslaw
authors.initials: M
authors.lastname: Kornek
mesh_terms: D000328:Adult; D000368:Aged; D017304:Annexin A5 / Q000097:blood; D037263:Asialoglycoprotein Receptor / Q000097:blood; D051926:Basigin / Q000097:blood; D001650:Bile Duct Neoplasms / Q000097:blood* / Q000175:diagnosis; D014408:Biomarkers, Tumor / Q000097:blood; D006528:Carcinoma, Hepatocellular / Q000097:blood* / Q000175:diagnosis; D045744:Cell Line, Tumor; D055252:Cell-Derived Microparticles / Q000473:pathology*; D018281:Cholangiocarcinoma / Q000097:blood* / Q000175:diagnosis; D003937:Diagnosis, Differential; D000071858:Epithelial Cell Adhesion Molecule / Q000097:blood; D005260:Female; D056945:Hep G2 Cells; D006801:Humans; D008113:Liver Neoplasms / Q000097:blood* / Q000175:diagnosis; D008297:Male; D008875:Middle Aged; D047368:Tumor Burden; D055815:Young Adult
chemical_list: C546927:ASGR1 protein, human; D017304:Annexin A5; D037263:Asialoglycoprotein Receptor; C497886:BSG protein, human; D014408:Biomarkers, Tumor; C499379:EPCAM protein, human; D000071858:Epithelial Cell Adhesion Molecule; D051926:Basigin
keywords: ASGPR1; Biomarker; CCA; CD147; CD326; Cancer; EMMPRIN; EpCAM; Extracellular vesicles; HCC; Microparticles; Microvesicles