Evaluation of AGP Fucosylation as a Marker for Hepatocellular Carcinoma of Three Different Etiologies

Jing Liang,Amit G Singal,Neehar D Parikh,Suyu Liu,Jianhui Zhu,Mobolaji Odewole,David M Lubman,Mengmeng Wang,Sofia Kagan,Veronica Renteria

doi:10.1038/s41598-019-48043-1

Abstract

A mass spectrometric analysis platform has been developed to determine whether glycosylation patterns of alpha-1 acid glycoprotein (AGP) could be used as a marker for early detection of hepatocellular carcinoma (HCC) in different etiologies, i.e. non-alcoholic steatohepatitis (NASH), alcoholic liver disease (ALC), and hepatitis C virus (HCV). MALDI-MS profiling of N-glycans of AGP purified from 20 μL of patient serum in HCC (n = 72) and liver cirrhosis (n = 58) showed that a unique trifucosylated tetra-antennary glycan (m/z 3490.76) was predominantly identified in HCCs but was absent in healthy subjects and the majority of cirrhosis patients. Receiver operation characteristic (ROC) curve analysis showed that the trifucosylated N-glycan of AGP (triFc_AGP) could differentiate HCC from cirrhosis with an area under the curve (AUC) of 0.707, 0.726 and 0.751 for NASH, ALC and HCV, respectively. When combining triFc_AGP with INR and AFP, the panel had the greatest benefit in detection of NASH-related HCCs, with a significantly improved AUC of 0.882 for all NASH HCCs and 0.818 for early NASH HCCs compared to AFP alone (0.761 and 0.641, respectively). Moreover, triFc_AGP could serve as a potential marker for monitoring AFP-negative HCC patients.

Highlights

Guidelines from the American Association for the Study of Liver Disease (AASLD) recommend hepatocellular carcinoma (HCC) surveillance with abdominal ultrasound every 6 months with or without alpha fetoprotein (AFP) in patients with cirrhosis[7]
Changes in serum acid glycoprotein (AGP) glycosylation are highly correlated with HCC progression[19,20,21]; it is unclear how N-glycan structures of AGP alter from liver cirrhosis to early HCC in different etiologies
We performed a comprehensive comparison of N-glycan structures of serum AGP between HCC and cirrhosis patients of the three common etiologies (i.e. non-alcoholic steatohepatitis (NASH), alcoholic liver disease (ALC), and hepatitis C virus (HCV)) in order to determine whether glycosylation patterns of AGP could be used as a marker for early detection of HCC in different etiologies

Summary

Introduction

Guidelines from the American Association for the Study of Liver Disease (AASLD) recommend HCC surveillance with abdominal ultrasound every 6 months with or without alpha fetoprotein (AFP) in patients with cirrhosis[7]. Aberrant glycosylation of AGP is highly related to the carcinoma progression of HCC19–21, especially with elevated fucosylation[22] It is still not clear how AGP glycoforms alter during the progression from liver cirrhosis to HCC of different etiologies. Zhang et al reported increased levels of fucosylation in AGP in patients with liver cirrhosis and HCC compared to healthy controls, where different degrees of fucosylation could distinguish HCC from cirrhosis[21]. These studies indicated that the fucosylation pattern of AGP is unique in patients with HCC, determination of the specific changes in glycan structures in AGP could become a sensitive biomarker for detecting early HCC. A comparison of different glycoforms of AGP characterized by MALDI-MS would provide information about its potential value as a biomarker

Methods

Results

Conclusion