title: Multiplex Serum Protein Analysis Identifies Novel Biomarkers of Advanced Fibrosis in Patients with Chronic Liver Disease with the Potential to Improve Diagnostic Accuracy of Established Biomarkers.
abstract: OBJECTIVE
  Non-invasive markers of liver fibrosis are urgently required, especially for use in non-specialist settings. The aim of this study was to identify novel serum biomarkers of advanced fibrosis.
  
  
  METHODS
  We performed an unbiased screen of 120 serum analytes including cytokines, chemokines and proteases in 70 patients (35 without fibrosis, 35 with cirrhosis on biopsy), and selected a panel of 44 candidate biomarkers, which were subsequently measured in a mixed-etiology cohort of 432 patients with known serum HA, PIIINP and TIMP1 (which comprise the validated Enhanced Liver Fibrosis (ELF) test). Multivariate logistic regression modelling was used to generate models for the prediction of advanced or significant fibrosis (METAVIR ≥F3 and ≥F2, respectively); in addition to identifying biomarkers of disease activity and steatohepatitis.
  
  
  RESULTS
  Seventeen analytes were significantly differentially expressed between patients with no advanced fibrosis and patients with advanced fibrosis, the most significant being hyaluronic acid (HA) and matrix metalloproteinase (MMP) 7 (p = 2.9E-41 and p = 1.0E-26, respectively). The optimal model for the prediction of advanced fibrosis comprised HA, MMP7, MMP1, alphafetoprotein (AFP) and the AST to platelet ratio index (APRI). We demonstrate enhanced diagnostic accuracy (AUROC = 0.938) compared to a model comprising HA, PIIINP and TIMP1 alone (ELF) (AUROC = 0.898, p<0.0001, De Long's test).
  
  
  CONCLUSIONS
  We have identified novel serum biomarkers of advanced liver fibrosis, which have the potential to enhance the diagnostic accuracy of established biomarkers. Our data suggest MMP7 is a valuable indicator of advanced fibrosis and may play a role in liver fibrogenesis.
authors.affiliation: Centre for Liver Disease Research, School of Medicine, The University of Queensland, Brisbane, Australia.
authors.forename: Katharine M
authors.initials: KM
authors.lastname: Irvine
authors.affiliation: Statistics Unit, QIMR Berghofer Medical Research Institute, Brisbane, Australia.
authors.forename: Leesa F
authors.initials: LF
authors.lastname: Wockner
authors.affiliation: Statistics Unit, QIMR Berghofer Medical Research Institute, Brisbane, Australia.
authors.forename: Isabell
authors.initials: I
authors.lastname: Hoffmann
authors.affiliation: Centre for Liver Disease Research, School of Medicine, The University of Queensland, Brisbane, Australia.|Department of Gastroenterology and Hepatology, Princess Alexandra Hospital, Brisbane, Australia.
authors.forename: Leigh U
authors.initials: LU
authors.lastname: Horsfall
authors.affiliation: Centre for Liver Disease Research, School of Medicine, The University of Queensland, Brisbane, Australia.
authors.forename: Kevin J
authors.initials: KJ
authors.lastname: Fagan
authors.affiliation: Institute of Molecular and Cell Biology, Agency for Science, Technology and Research, Singapore 138673; Singapore Immunology Network, Singapore 138648; and Institute of Biomedical Studies, Baylor University, Waco, Texas, 76798, United States of America.
authors.forename: Veonice
authors.initials: V
authors.lastname: Bijin
authors.affiliation: Institute of Molecular and Cell Biology, Agency for Science, Technology and Research, Singapore 138673; Singapore Immunology Network, Singapore 138648; and Institute of Biomedical Studies, Baylor University, Waco, Texas, 76798, United States of America.
authors.forename: Bernett
authors.initials: B
authors.lastname: Lee
authors.affiliation: Centre for Liver Disease Research, School of Medicine, The University of Queensland, Brisbane, Australia.
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Clouston
authors.affiliation: Pathology Queensland, Princess Alexandra Hospital, Brisbane, Australia.
authors.forename: Guy
authors.initials: G
authors.lastname: Lampe
authors.affiliation: Institute of Molecular and Cell Biology, Agency for Science, Technology and Research, Singapore 138673; Singapore Immunology Network, Singapore 138648; and Institute of Biomedical Studies, Baylor University, Waco, Texas, 76798, United States of America.
authors.forename: John E
authors.initials: JE
authors.lastname: Connolly
authors.affiliation: Centre for Liver Disease Research, School of Medicine, The University of Queensland, Brisbane, Australia.|Department of Gastroenterology and Hepatology, Princess Alexandra Hospital, Brisbane, Australia.
authors.forename: Elizabeth E
authors.initials: EE
authors.lastname: Powell
mesh_terms: D000328:Adult; D015415:Biomarkers / Q000097:blood*; D001798:Blood Proteins; D003430:Cross-Sectional Studies; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D008103:Liver Cirrhosis / Q000097:blood* / Q000175:diagnosis*; D016015:Logistic Models; D008297:Male; D008875:Middle Aged; D015203:Reproducibility of Results; D012189:Retrospective Studies; D012720:Severity of Illness Index
chemical_list: D015415:Biomarkers; D001798:Blood Proteins