Plasma DNA methylation marker and hepatocellular carcinoma risk prediction model for the general population

Abstract

Metastases in the later stages of hepatocellular carcinoma (HCC) cause the majority of deaths associated with the disease, making early detection crucial to patient survival. Risk models assessing HCC risk in the general population can be used for risk stratification for further HCC surveillance, however, none have been validated externally. Methylation of circulating DNA shows potential for non-invasive diagnosis of HCC. We conducted a prospective case-control study nested within a community-based cohort. We measured methylation levels in six genes (CDKN2A, RASSF1A, STEAP4, TBX2, VIM and ZNF154) which were identified in our previous work, using pre-diagnostic plasma DNA from 237 HCC cases and 257 matched controls. We found TBX2 hypermethylation was associated with increased HCC risk, with ORs (95% CI) of 3.2 (1.8-6.0). The associations were mainly among high-risk subjects; among subjects infected with HBV/HCV, the OR (95% CI) of TBX2 methylation was 5.3 (2.2-12.7). Among subjects with high risk scores, the ORs (95% CIs) were 7.8 (1.5-38.6) for Wen-HCC model ≥16, 5.8 (2.2-15.5) for Hung-HCC ≥15 and 7.5 (2.2-26.0) for Michikawa-HCC ≥8. Adding TBX2 methylation improved the accuracy of risk models for a high-risk population, with the area under the curve (AUC) of 76% for Wen-HCC score with TBX2 methylation compared with 69% with Wen-HCC alone. The AUCs were 63% for Hung-HCC score plus TBX2 methylation, and 53% for Hung-HCC alone, 65% for Michikawa-HCC score plus TBX2 methylation and 58% for Michikawa-HCC alone. Our findings suggest the potential increase in risk assessment discrimination and accuracy from incorporation of DNA methylation.

Identification of RASSF1A Promoter Hypermethylation as a Biomarker for Hepatocellular Carcinoma

“…rst reported that the promoter hypermethylation of RASSF1A was a major inactivating event in 85% (70/82) of HCC patients [18]. Since then, numerous studies with small sample sizes have demonstrated that the rate of RASSF1A promoter methylation is signi cantly increased within HCC tissues relative to noncarcinoma tissues [36,44,59], and the results of association analyses were consistent across studies [26,35,57]. Then, Zhao et al conducted a meta-analysis that involved a total of seven case-control studies, which suggested that the promoter hypermethylation of RASSF1A within body uids was signi cantly correlated with HCC risk [11].…”

Identification of RASSF1A Promoter Hypermethylation as a Biomarker for Hepatocellular Carcinoma

“…Some studies suggested that both radiotherapy and chemotherapy can alter a patient's DNA methylation status; therefore, the type of preoperative treatment performed should be clari ed [83,84]. Moreover, four studies investigated the relationship between RASSF1A hypermethylation in peripheral blood and the risk of HCC, and the diagnosis of HCC was con rmed by imaging techniques and serum AFP levels, rather than through pathological examination [56][57][58][59]. The diagnosis of HCC based on imaging techniques is recommended by guidelines; however, histopathological evaluation remains the gold standard for HCC diagnosis [85,86].…”

Impact of circulating tumor DNA in hepatocellular and pancreatic carcinomas

“…Xu et al constructed a diagnostic prediction model using a cfDNA methylation marker panel that predicted HCC survival and could effectively discriminate patients with HCC from individuals with HBV/ HCV infection, fatty liver disease as well as healthy controls superior to AFP (Xu et al 2017). Although a multitude of aberrant methylated genes could be identified as prognostic target in HCC, there is no recognized biomarker confirmed in multiple centers (Han et al 2014;Iizuka et al 2011;Ji et al 2014;Oussalah et al 2018;Sun et al 2013;Wang et al 2006;Wen et al 2015;Wu et al 2017;Zhang et al 2013).…”