The development of a cSMART-based integrated model for hepatocellular carcinoma diagnosis

Abstract

Background: Hepatocellular carcinoma (HCC) generally arises from a background of liver cirrhosis (LC). Patients with cirrhosis and suspected HCC are recommended to undergo serum biomarker tests and imaging diagnostic evaluation. However, the performance of routine diagnostic methods in detecting early HCC remains unpromising.

Methods: Here, we conducted a large-scale, multicenter study of 1675 participants including 490 healthy controls, 577 LC patients, and 608 HCC patients from nine clinical centers across nine provinces of China, profiled gene mutation signatures of cell-free DNA (cfDNA) using Circulating Single-Molecule Amplification and Resequencing Technology (cSMART) through detecting 931 mutation sites across 21 genes.

Results: An integrated diagnostic model called "Combined method" was developed by combining three mutation sites and three serum biomarkers. Combined method outperformed AFP in the diagnosis of HCC, especially early HCC, with sensitivities of 81.25% for all stages and 66.67% for early HCC, respectively. Importantly, the integrated model exhibited high accuracy in differentiating AFP-negative, AFP-L3-negative, and PIVKA-II-negative HCCs from LCs.

Keywords: Biomarker; Cell-free DNA; Diagnosis; Hepatocellular carcinoma; Mutation.

Fig. 1

Combined method holds a strong value in diagnosis of HCC. a Basic information (age, gender), cirrhosis background, tumor serological biomarkers (CA199, PIVKA-II, AFP-L3, AFP), and HCC related parameters (MVI and BCLC stages) of all HCC samples with positive mutations at the top 20 high-frequency mutation sites. CA199: carbohydrate antigen199; MVI: microvascular invasion; and BCLC: Barcelona clinic liver cancer. b ROC curves of Combined method and AFP for HCC patients versus LC patients in the training, test, and validation cohorts. c Proportions of positive and negative calling by Combined method, GALAD, and AFP in all participants with different AFP, AFP-L3, and PIVKA-II levels in test cohort. d Proportions of positive and negative calling by Combined method, GALAD, and AFP in all participants with different age, gender, Child–Pugh stages, HBV infection status, and cirrhosis history in test cohort. e Proportions of positive and negative calling by Combined method, GALAD, and AFP in HCC patients with different tumor sizes and BCLC stages in test cohort