Circulating Tumor DNA Analysis for Liver Cancers and Its Usefulness as a Liquid Biopsy
Atsushi Ono, Akihiro Fujimoto, Yujiro Yamamoto, Sakura Akamatsu, Nobuhiko Hiraga, Michio Imamura, Tomokazu Kawaoka, Masataka Tsuge, Hiromi Abe, C Nelson Hayes, Daiki Miki, Mayuko Furuta, Tatsuhiko Tsunoda, Satoru Miyano, Michiaki Kubo, Hiroshi Aikata, Hidenori Ochi, Yoshi-Iku Kawakami, Koji Arihiro, Hideki Ohdan, Hidewaki Nakagawa, Kazuaki Chayama, Atsushi Ono, Akihiro Fujimoto, Yujiro Yamamoto, Sakura Akamatsu, Nobuhiko Hiraga, Michio Imamura, Tomokazu Kawaoka, Masataka Tsuge, Hiromi Abe, C Nelson Hayes, Daiki Miki, Mayuko Furuta, Tatsuhiko Tsunoda, Satoru Miyano, Michiaki Kubo, Hiroshi Aikata, Hidenori Ochi, Yoshi-Iku Kawakami, Koji Arihiro, Hideki Ohdan, Hidewaki Nakagawa, Kazuaki Chayama
Abstract
Background & aims: Circulating tumor DNA (ctDNA) carrying tumor-specific sequence alterations has been found in the cell-free fraction of blood. Liver cancer tumor specimens are difficult to obtain, and noninvasive methods are required to assess cancer progression and characterize underlying genomic features.
Methods: We analyzed 46 patients with hepatocellular carcinoma who underwent hepatectomy or liver transplantation and for whom whole-genome sequencing data was available. We designed personalized assays targeting somatic rearrangements of each tumor to quantify serum ctDNA. Exome sequencing was performed using cell-free DNA paired primary tumor tissue DNA from a patient with recurrent liver cancer after transcatheter arterial chemoembolization (TACE).
Results: We successfully detected ctDNA from 100 μL of serum samples in 7 of the 46 patients before surgery, increasing with disease progression. The cumulative incidence of recurrence and extrahepatic metastasis in the ctDNA-positive group were statistically significantly worse than in the ctDNA-negative group (P = .0102 and .0386, respectively). Multivariate analysis identified ctDNA (OR 6.10; 95% CI, 1.11-33.33, P = .038) as an independent predictor of microscopic vascular invasion of the portal vein (VP). We identified 45 nonsynonymous somatic mutations in cell-free DNA after TACE and 71 nonsynonymous somatic mutations in primary tumor tissue by exome sequencing. We identified 25 common mutations in both samples, and 83% of mutations identified in the primary tumor could be detected in the cell-free DNA.
Conclusions: The presence of ctDNA reflects tumor progression, and detection of ctDNA can predict VP and recurrence, especially extrahepatic metastasis within 2 years. Our study demonstrated the usefulness of ctDNA detection and sequencing analysis of cell-free DNA for personalized treatment of liver cancer.
Keywords: AFP, α-fetoprotein; ALT, alanine aminotransferase; AST, aspartate aminotransferase; Circulating Tumor DNA; DCP, des-γ-carboxy prothrombin; Exome Sequencing; HAIC, hepatic arterial infusion chemotherapy; HBV, hepatitis B virus; HCC, hepatocellular carcinoma; HCV, hepatitis C virus; Hepatocellular Carcinoma; PCR, polymerase-chain-reaction; TACE, transcatheter arterial chemoembolization; VP, microscopic vascular invasion to portal vein; Whole-Genome Sequencing; cHCC/CC, combined hepatocellular and cholangiocarcinoma; ctDNA, circulating tumor DNA.
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