Currently available diagnostic tools for HCC lack sensitivity and specificity and deliver little value to patients. We investigated the use of circulating serum glycoproteins to identify a panel of potential prognostic markers that may be indicative of progression from healthy state to NASH and further to HCC.
Fatty liver disease progresses through stages of fat accumulation and inflammation to non-alcoholic steatohepatitis (NASH), fibrosis and cirrhosis and eventually hepatocellular carcinoma (HCC). Currently available diagnostic tools for HCC lack sensitivity and specificity and deliver little value to patients. In this study, we investigated the use of circulating serum glycoproteins to identify a panel of potential prognostic markers that may be indicative of progression from the healthy state to NASH and further to HCC. Serum samples were processed using a standard preanalytical sample preparation protocol and were analyzed using a novel high throughput glycoproteomics platform. Relative abundance of 413 glycopeptides, representing 57 abundant serum proteins were determined and compared among the three phenotypes. We used PB-net, a peak picking software built in-house, to quantify area under the peaks. Our initial dataset, containing healthy, NASH, and HCC serum samples, yielded several glycopeptides that demon-strated statistically significant differences in abundances in NASH and HCC compared to con-trols. We analyzed the relative abundance of common glycoforms and observed higher levels of core-fucosylated, sialylated and branched glycans, in NASH and HCC as compared to controls. We replicated these findings in an independent set of samples of individuals with benign liver conditions and HCC, respectively. Glycoproteomic analysis of serum proteins is a novel source of prognostic biomarkers differentially associated with absence of liver disease vs. NASH vs. HCC, respectively. Our results may be of value in the management of patients with liver disease.