Plasma methylated GNB4 and Riplet as a novel dual-marker panel for the detection of hepatocellular carcinoma

Abstract

Early detection of hepatocellular carcinoma (HCC) can greatly improve the survival rate of patients. We aimed to develop a novel marker panel based on cell-free DNA (cfDNA) methylation for the detection of HCC. The differentially methylated CpG sites (DMCs) specific for HCC blood diagnosis were selected from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases, then validated by the whole genome bisulphite sequencing (WGBS) of 12 paired HCC and paracancerous tissues. The clinical performance of the panel was evaluated using tissue samples [32 HCC, chronic liver disease (CLD), and healthy individuals] and plasma cohorts (173 HCC, 199 CLD, and 98 healthy individuals). The combination of G protein subunit beta 4 (GNB4) and Riplet had the optimal area under the curve (AUC) in seven candidates through TCGA, GEO, and WGBS analyses. In tissue validation, the GNB4 and Riplet showed an AUC of 100% with a sensitivity and specificity of 100% for detecting any-stage HCC. In plasma, it demonstrated a high sensitivity of 84.39% at 91.92% specificity, with an AUC of 92.51% for detecting any-stage HCC. The dual-marker panel had a higher sensitivity of 78.26% for stage I HCC than alpha-fetoprotein (AFP) of 47.83%, and a high sensitivity of 70.27% for detecting a single tumour (size ≤3 cm). In conclusion, we developed a novel dual-marker panel that demonstrates high accuracy in detecting HCC, surpassing the performance of AFP testing.

A dual-gene panel of two fragments of methylated IRF4 and one of ZEB2 in plasma cell-free DNA for gastric cancer detection

“…Therefore, healthy white blood cells were used as controls in tissue validation for this study. They were also used as controls in other studies [ 42 , 46 , 47 ].…”

Research Progress on the Role of Epigenetic Methylation Modification in Hepatocellular Carcinoma

“…Several studies have shown that combining Cell-free DNA (cfDNA) methylation-specific high-throughput or MSP testing with serology enhances the accuracy of serological assays of alpha-fetoprotein (AFP) alone during early screening for HCC. 40 …”

“…Several studies have shown that combining Cell-free DNA (cfDNA) methylation-specific highthroughput or MSP testing with serology enhances the accuracy of serological assays of alpha-fetoprotein (AFP) alone during early screening for HCC. 40 In targeted therapy of HCC, DNA methyltransferases (DNMT) and DNA demethylases (ten-eleven translocation, TET) in methylation modification may play an important role. In cancer stem-like cells (CSCs), coordinated activation of DNMT3a and TET2 triggers and maintains drug resistance, which would be a therapeutic target for refractory HCC.…”

An AgPd NP-based lateral flow immunoassay for simultaneous detection of glycocholic acid and alpha-fetoprotein

“…1 New cases of HCC in China, a high-risk country for HCC, accounted for 45.3% of all new HCC cases worldwide in 2022. 2 Early detection of liver diseases is crucial, as it correlates with an enhanced 5 year relative survival rate and prognosis. 3 In clinical practice, serum biomarkers such as aspartate aminotransferase (AST), alkaline phosphatase (ALP), and alanine aminotransferase (ALT) are commonly employed to reflect the pathological status of the liver, which is the main method for liver function detection at present.…”