title: Mass-Selected Site-Specific Core-Fucosylation of Serum Proteins in Hepatocellular Carcinoma.
abstract: A mass spectrometry-based methodology has been developed to screen for changes in site-specific core-fucosylation (CF) of serum proteins in early stage HCC with different etiologies. The methods involve depletion of high-abundance proteins, trypsin digestion of medium-to-low-abundance proteins into peptides, iTRAQ labeling, and Lens culinaris Agglutinin (LCA) enrichment of CF peptides, followed by endoglycosidase F3 digestion before mass spectrometry analysis. 1300 CF peptides from 613 CF proteins were identified from patients sera, where 20 CF peptides were differentially expressed in alcohol (ALC)-related HCC samples compared with ALC-related cirrhosis samples and 26 CF peptides changed in hepatitis C virus (HCV)-related HCC samples compared with HCV-related cirrhosis samples. Among these, we found three CF peptides from fibronectin upregulated in ALC-related HCC samples compared with ALC-related cirrhosis samples with an AUC (area under the curve) value of 0.89 at site 1007 with a specificity of 85.7% at a sensitivity of 92.9% (generated with 10-fold cross-validation). When combined with the AFP index, the AUC value reached to 0.92 with a specificity of 92.9% at a sensitivity of 100%, significantly improved compared to that with AFP alone (LR test p < 0.001). In HCV-related samples, the CF level of cadherin-5 at site 61 showed the best AUC value of 0.75 but was not as promising as that of fibronectin site 1007 for ALC-related samples. The CF peptides of fibronectin may serve as potential biomarkers for early stage HCC screening in ALC-related cirrhosis patients.
authors.affiliation: Department of Surgery, University of Michigan Medical Center , Ann Arbor, Michigan 48109, United States.
authors.forename: Haidi
authors.initials: H
authors.lastname: Yin
authors.affiliation: Department of Surgery, University of Michigan Medical Center , Ann Arbor, Michigan 48109, United States.
authors.forename: Zhijing
authors.initials: Z
authors.lastname: Tan
authors.affiliation: Department of Surgery, University of Michigan Medical Center , Ann Arbor, Michigan 48109, United States.
authors.forename: Jing
authors.initials: J
authors.lastname: Wu
authors.affiliation: Department of Surgery, University of Michigan Medical Center , Ann Arbor, Michigan 48109, United States.
authors.forename: Jianhui
authors.initials: J
authors.lastname: Zhu
authors.affiliation: Department of Statistics, University of Michigan , Ann Arbor, Michigan 48109, United States.
authors.forename: Kerby A
authors.initials: KA
authors.lastname: Shedden
authors.affiliation: Liver Transplantation Program, University of Texas Southwestern Medical Center , Dallas, Texas 75390, United States.
authors.forename: Jorge
authors.initials: J
authors.lastname: Marrero
authors.affiliation: Department of Surgery, University of Michigan Medical Center , Ann Arbor, Michigan 48109, United States.
authors.forename: David M
authors.initials: DM
authors.lastname: Lubman
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D000437:Alcoholism / Q000097:blood* / Q000150:complications / Q000235:genetics / Q000473:pathology; D000595:Amino Acid Sequence; D015703:Antigens, CD / Q000097:blood / Q000235:genetics; D015415:Biomarkers / Q000097:blood; D001798:Blood Proteins / Q000235:genetics / Q000378:metabolism*; D015820:Cadherins / Q000097:blood / Q000235:genetics; D006528:Carcinoma, Hepatocellular / Q000097:blood* / Q000209:etiology / Q000235:genetics / Q000473:pathology; D005260:Female; D005353:Fibronectins / Q000097:blood / Q000235:genetics; D005643:Fucose / Q000378:metabolism; D006509:Hepatitis B / Q000097:blood* / Q000150:complications / Q000235:genetics / Q000473:pathology; D006526:Hepatitis C / Q000097:blood* / Q000150:complications / Q000235:genetics / Q000473:pathology; D006801:Humans; D008103:Liver Cirrhosis / Q000097:blood* / Q000150:complications / Q000235:genetics / Q000473:pathology; D008113:Liver Neoplasms / Q000097:blood* / Q000209:etiology / Q000235:genetics / Q000473:pathology; D008297:Male; D008875:Middle Aged; D058977:Molecular Sequence Annotation; D008969:Molecular Sequence Data; D009367:Neoplasm Staging; D010449:Peptide Mapping; D011499:Protein Processing, Post-Translational; D059748:Proteolysis; D014357:Trypsin / Q000737:chemistry; D000099152:Cadherin 5
chemical_list: D015703:Antigens, CD; D015415:Biomarkers; D001798:Blood Proteins; D015820:Cadherins; D005353:Fibronectins; D005643:Fucose; D014357:Trypsin; D000099152:Cadherin 5
keywords: Core-fucosylation; alcohol-related HCC; hepatitis C virus-related HCC; hepatocellular carcinoma; site-specific