PLoS ONE: Discovery of Novel Biomarker Candidates for Liver Fibrosis in Hepatitis C Patients: A Preliminary Study

Posted 7/5/12 on PLoS ONE.org. UK researchers utilized methodology within the discipline of Proteomics to identify plasma-based proteins that could possibly be used clinically to discriminate between the intermediate stages of fibrosis. Twenty biomarkers were validated in comparison to the commonly used markers in FibroTest, ELF, Hepascore and FIBROSpect by Western blotting. These markers will be further validated using a large clinical cohort. Any advance in this area that would possibly be used to avoid liver biopsy would greatly benefit patients. 


Discovery of Novel Biomarker Candidates for Liver Fibrosis in Hepatitis C Patients: A Preliminary Study

Bevin Gangadharan1*, Manisha Bapat1#, Jan Rossa1#, Robin Antrobus1, David Chittenden1, Bettina Kampa1, Eleanor Barnes2,3, Paul Klenerman2, Raymond A. Dwek1, Nicole Zitzmann1
1 Oxford Antiviral Drug Discovery Unit, Oxford Glycobiology Institute, Department of Biochemistry, University of Oxford, Oxford, United Kingdom, 2 Nuffield Department of Clinical Medicine, University of Oxford, Oxford, United Kingdom, 3 Oxford NIHR Biomedical Research Centre, The John Radcliffe Hospital, Headington, Oxford, United Kingdom

Abstract

Background
Liver biopsy is the reference standard for assessing liver fibrosis and no reliable non-invasive diagnostic approach is available to discriminate between the intermediate stages of fibrosis. Therefore suitable serological biomarkers of liver fibrosis are urgently needed. We used proteomics to identify novel fibrosis biomarkers in hepatitis C patients with different degrees of liver fibrosis.

Methodology/Principal Findings
Proteins in plasma samples from healthy control individuals and patients with hepatitis C virus (HCV) induced cirrhosis were analysed using a proteomics technique: two dimensional gel electrophoresis (2-DE). This technique separated the proteins in plasma samples of control and cirrhotic patients and by visualizing the separated proteins we were able to identify proteins which were increasing or decreasing in hepatic cirrhosis. Identified markers were validated across all Ishak fibrosis stages and compared to the markers used in FibroTest, Enhanced Liver Fibrosis (ELF) test, Hepascore and FIBROSpect by Western blotting. Forty four candidate biomarkers for hepatic fibrosis were identified of which 20 were novel biomarkers of liver fibrosis. Western blot validation of all candidate markers using plasma samples from patients across all Ishak fibrosis scores showed that the markers which changed with increasing fibrosis most consistently included lipid transfer inhibitor protein, complement C3d, corticosteroid-binding globulin, apolipoprotein J and apolipoprotein L1. These five novel fibrosis markers which are secreted in blood showed a promising consistent change with increasing fibrosis stage when compared to the markers used for the FibroTest, ELF test, Hepascore and FIBROSpect. These markers will be further validated using a large clinical cohort.

Conclusions/Significance
This study identifies 20 novel fibrosis biomarker candidates. The proteins identified may help to assess hepatic fibrosis and eliminate the need for invasive liver biopsies.

Citation: Gangadharan B, Bapat M, Rossa J, Antrobus R, Chittenden D, et al. (2012) Discovery of Novel Biomarker Candidates for Liver Fibrosis in Hepatitis C Patients: A Preliminary Study. PLoS ONE 7(6): e39603. doi:10.1371/journal.pone.0039603

Editor: Ravi Jhaveri, Duke University School of Medicine, United States of America

Received: November 30, 2011; Accepted: May 22, 2012; Published: June 26, 2012

Copyright: © 2012 Gangadharan et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Funding: This work was supported by the Oxford Glycobiology Endowment and a ‘Blue Skies’ research grant from United Therapeutics Corp. NZ is a Senior Research Fellow of Linacre College, Oxford. PK was supported by the Wellcome Trust, The James Martin School for the 21st Century and the NIHR Biomedical Research Centre Programme (Oxford). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Competing interests: Professor Raymond Dwek is a Director and Member of the Scientific Board of United Therapeutics Corp., which supported this work in part through a “Blue Skies” research grant. A patent covering the biomarkers in this study has been filed. The patent number is 61178334. This does not alter the authors' adherence to all the PLoS ONE policies on sharing data and materials.

* E-mail: Bevin.Gangadharan@bioch.ox.ac.uk

# These authors contributed equally to this work.