Detection and Analysis of Circulating Epithelial Cells in Liquid Biopsies From Patients With Liver Disease

Abstract Epithelial cells in the circulation (circulating epithelial cells, or CECs) are analyzed as a non-invasive method to detect cancers; we investigated whether analysis of hepatocytes in the circulation can identify patients with chronic liver disease or hepatocellular carcinoma (HCC). We previously developed a cell-sorting device to isolate CECs from patient blood samples and combined it with an mRNA analysis system to identify CECs with liver-specific markers. We tested the ability of this device to detect CECs of hepatocyte origin in blood samples from healthy individuals (n=10), patients with chronic liver disease without HCC (n=39), and patients with HCC (n=54), using immunofluorescence. We confirmed hepatocyte origin using RNA-sequencing analysis. We found a similar concentration of circulating hepatocytes in blood samples from patients with chronic liver disease and HCC but an increased concentration from patients with advanced fibrosis compared to those without advanced fibrosis. Circulating hepatocytes isolated from patients with HCC had a different gene expression profile than those from patients with chronic liver disease. This system for detecting and analyzing circulating hepatocytes might be used in the evaluation of benign and malignant liver disease.