Biomarker discovery for early detection of hepatocellular carcinoma in hepatitis C-Infected patients

Mehnaz G. Mustafa, John R. Petersen, Hyunsu Ju, Luca Cicalese, Ned Snyder, Sigmund J. Haidacher, Larry Denner, Cornelis Elferink

Research output: Contribution to journalArticle

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Abstract

Chronic hepatic disease damages the liver, and the resulting wound-healing process leads to liver fibrosis and the subsequent development of cirrhosis. The leading cause of hepatic fibrosis and cirrhosis is infection with hepatitis C virus (HCV), and of the patients with HCVinduced cirrhosis, 2% to 5% develop hepatocellular carcinoma (HCC), with a survival rate of 7%. HCC is one of the leading causes of cancer-related death worldwide, and the poor survival rate is largely due to late-stage diagnosis, which makes successful intervention difficult, if not impossible. The lack of sensitive and specific diagnostic tools and the urgent need for early-stage diagnosis prompted us to discover new candidate biomarkers for HCV and HCC. We used aptamer-based fractionation technology to reduce serum complexity, differentially labeled samples (six HCV and six HCC) with fluorescent dyes, and resolved proteins in pairwise two-dimensional difference gel electrophoresis. DeCyder software was used to identify differentially expressed proteins and spots picked, and MALDI-MS/MS was used to determine that ApoA1 was down-regulated by 22% (p < 0.004) in HCC relative to HCV. Differential expression quantified via two-dimensional difference gel electrophoresis was confirmed by means of 18O/ 16O stable isotope differential labeling with LC-MS/MS zoom scans. Technically independent confirmation was demonstrated by triple quadrupole LC-MS/MS selected reaction monitoring (SRM) assays with three peptides specific to human ApoA1 (DLATVYVDVLK, WQEEMELYR, and VSFLSALEEYTK) using 18O/16O-labeled samples and further verified with AQUA peptides as internal standards for quantification. In 50 patient samples (24 HCV and 26 HCC), all three SRM assays yielded highly similar differential expression of ApoA1 in HCC and HCV patients. These results validated the SRM assays, which were independently confirmed by Western blotting. Thus, ApoA1 is a candidate member of an SRM biomarker panel for early diagnosis, prognosis, and monitoring of HCC. Future multiplexing of SRM assays for other candidate biomarkers is envisioned to develop a biomarker panel for subsequent verification and validation studies. Molecular & Cellular Proteomics 12: 10.1074/mcp.M113.031252, 3640-3652, 2013.

Original languageEnglish (US)
Pages (from-to)3640-3652
Number of pages13
JournalMolecular and Cellular Proteomics
Volume12
Issue number12
DOIs
StatePublished - Dec 2013

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Biomarkers
Hepatitis C
Viruses
Hepatocellular Carcinoma
Hepacivirus
Monitoring
Assays
Two-Dimensional Difference Gel Electrophoresis
Electrophoresis
Liver
Fibrosis
Gels
Liver Cirrhosis
Early Diagnosis
Peptides
Survival Rate
Fractionation
Isotope Labeling
Fluorescent Dyes
Multiplexing

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Analytical Chemistry

Cite this

Biomarker discovery for early detection of hepatocellular carcinoma in hepatitis C-Infected patients. / Mustafa, Mehnaz G.; Petersen, John R.; Ju, Hyunsu; Cicalese, Luca; Snyder, Ned; Haidacher, Sigmund J.; Denner, Larry; Elferink, Cornelis.

In: Molecular and Cellular Proteomics, Vol. 12, No. 12, 12.2013, p. 3640-3652.

Research output: Contribution to journalArticle

Mustafa, Mehnaz G. ; Petersen, John R. ; Ju, Hyunsu ; Cicalese, Luca ; Snyder, Ned ; Haidacher, Sigmund J. ; Denner, Larry ; Elferink, Cornelis. / Biomarker discovery for early detection of hepatocellular carcinoma in hepatitis C-Infected patients. In: Molecular and Cellular Proteomics. 2013 ; Vol. 12, No. 12. pp. 3640-3652.
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