LC/MS-Based Global Metabolomic Identification of Serum Biomarkers Differentiating Hepatocellular Carcinoma from Chronic Hepatitis B and Liver Cirrhosis

Abstract

Chronic hepatitis B virus (CHB) infection is one of the primary risk factors associated with the development of hepatocellular carcinoma (HCC). Despite having been extensively studied, diagnosing early-stage HCC remains challenging, and diagnosed patients have a poor (3–5%) survival rate. Identifying new approaches to detect changes in the serum metabolic profiles of patients with CHB and liver cirrhosis (LC) may provide a valuable approach to better detect HCC at an early stage when it is still amenable to treatment, thereby improving patient prognosis and survival. In the present study, we, therefore, employed a liquid chromatography-mass spectrometry (LC-MS)-based approach to evaluate the serum metabolic profiles of 30 CHB patients, 29 LC patients, and 30 HCC patients. We then employed appropriate statistical methods to identify those metabolites that were best able to distinguish HCC cases from LC and CHB controls. A mass-based database was then used to putatively identify these metabolites. We then confirmed the identities of a subset of these metabolites through comparisons with the MS/MS fragmentation patterns and retention times of reference standards. The serum samples were then reanalyzed to quantify the levels of these selected metabolites and of other metabolites that have previously been identified as potential HCC biomarkers. Through this approach, we observed clear differences in the metabolite profiles of the CHB, LC, and HCC patient groups in both positive- and negative-ion modes. We found that the levels of taurodeoxy cholic acid (TCA) and 1,2-diacyl-3-β-d-galactosyl-sn-glycerol rose with the progression from CHB to LC to HCC, whereas levels of 5-hydroxy-6E,8Z,11Z,14Z,17Z-eicosapentaenoic acid, and glycyrrhizic acid were gradually reduced with liver disease progression in these groups. The ROC analysis showed that taurodeoxy cholic acid (TCA), 1,2-diacyl-3-β-d-galactosyl-sn-glycerol, 5-hydroxy-6E,8Z,11Z,14Z,17Z-eicosapentaenoic acid, and glycyrrhizic acid had a diagnosis performance with liver disease progression. These four metabolites have a significant correlation with alpha fetal protein (AFP) level and age. Our results highlight novel metabolic biomarkers that have the potential to be used for differentiating between CHB, LC, and HCC patients, thereby facilitating the identification and treatment of patients with early-stage HCC.

Metabolomics in hepatocellular carcinoma: From biomarker discovery to precision medicine

“…By comparing metabolite profiling from liver cirrhosis patients, 51 differential metabolites were selected, which were altered with the progression from chronic hepatitis B to liver cirrhosis to HCC. Specifically, the levels of taurodeoxy cholic acid and 1,2-diacyl-3-β-D-galactosyl-sn-glycerol were gradually increased with liver disease progression, whereas the levels of 5-hydroxy-6E,8Z,11Z,14Z,17Z-eicosapentaenoic acid and glycyrrhizic acid were decreased ( 88 ). As for HCV-based HCC, 40 HCC patients with underlying HCV infections and 22 HCV patients were recruited for a 1 H-NMR-based study.…”

Circulating Metabolic Markers Related to the Diagnosis of Hepatocellular Carcinoma

“…Serum bile acids can afect the tumor immune microenvironment (TIME) associated with the intestinal microbiota, which may have a signifcant impact on the tumor load and adverse clinical outcomes of HBV-associated HCC patients [77]. As the disease progresses from chronic viral hepatitis B to cirrhosis and then to HCC, the taurodeoxycholic acid level continues to increase while the glycyrrhizin acid level decreases [62]. Terefore, taurodeoxycholic acid and glycyrrhizin acids have the potential to distinguish patients with CHB, LC, and HCC, thereby facilitating the identifcation of patients with early HCC and access to curative therapy.…”

Hepatocellular Carcinoma in 2021: An Exhaustive Update

“…Furthermore, the panel was also found to have prowess in detecting AFP-negative HCC as well as solitary HCC nodules, or at most two nodules less than 3 cm in diameter. In another study, the levels of taurodeoxy cholic acid and 1,2-diacyl-3-β-d-galactosyl-sn-glycerol increased with the progression from chronic HBV-related liver disease to cirrhosis to HCC transformation, whereas the levels of 5-hydroxy-6E,8Z,11Z,14Z,17Z-eicosapentaenoic acid, and glycyrrhizic acid were found to gradually decrease with liver disease progression [ 65 - 66 ].…”