Detection of early-stage hepatocellular carcinoma in asymptomatic HBsAg-seropositive individuals by liquid biopsy

Abstract

Liquid biopsies, based on cell free DNA (cfDNA) and proteins, have shown the potential to detect early stage cancers of diverse tissue types. However, most of these studies were retrospective, using individuals previously diagnosed with cancer as cases and healthy individuals as controls. Here, we developed a liquid biopsy assay, named the hepatocellular carcinoma screen (HCCscreen), to identify HCC from the surface antigen of hepatitis B virus (HBsAg) positive asymptomatic individuals in the community population. The training cohort consisted of individuals who had liver nodules and/or elevated serum α-fetoprotein (AFP) levels, and the assay robustly separated those with HCC from those who were non-HCC with a sensitivity of 85% and a specificity of 93%. We further applied this assay to 331 individuals with normal liver ultrasonography and serum AFP levels. A total of 24 positive cases were identified, and a clinical follow-up for 6-8 mo confirmed four had developed HCC. No HCC cases were diagnosed from the 307 test-negative individuals in the follow-up during the same timescale. Thus, the assay showed 100% sensitivity, 94% specificity, and 17% positive predictive value in the validation cohort. Notably, each of the four HCC cases was at the early stage (<3 cm) when diagnosed. Our study provides evidence that the use of combined detection of cfDNA alterations and protein markers is a feasible approach to identify early stage HCC from asymptomatic community populations with unknown HCC status.

Keywords: HBsAg-seropositive; cell free DNA; early detection of cancer; hepatocellular carcinoma.

Conflict of interest statement

Conflict of interest statement: S.W. and H. Yan are the founders of Genetron Health (Beijing) Co. Ltd. Y.J. is one of the cofounders of Genetron Health (Beijing) Co. Ltd. B.H.D. is a consultant for Genetron Health (Beijing) Co. Ltd. Y.J., Q.C., P.W., Yuting Wang, K.C., Q.S., S.W., and H. Yan have filed patents/patent applications based on the data generated from this work. Other authors declare no conflict of interest.

Figures

Fig. 1.

Study design. The enrollment, training of the HCCscreen model, and validation in sampled AFP/US-negative individuals.

Fig. 2.

The performance of HCCscreen in the training and validation cohorts. (A) The HCCscreen scores and contributions of the cfDNA and protein biomarkers in the diagnostic model in the training cohort. (B) Binary results of the diagnostic model in the training cohort. (C) ROC of the diagnostic model of the HCCscreen in the training cohort. (D) The HCCscreen performance of the diagnostic model in the validation cohort. (E) Follow-up and diagnosis of the HCCscreen-positive cases in the validation cohort. (F) Binary results of the diagnostic model in the validation cohort. (G) Dynamic CT imaging of the four HCC cases detected with the HCCscreen among the AFP/US-negative individuals.