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Post-treatment cell-free DNA as a predictive biomarker in molecular-targeted therapy of hepatocellular carcinoma

  • Original Article―Liver, Pancreas, and Biliary Tract
  • Published:
Journal of Gastroenterology Aims and scope Submit manuscript

Abstract

Background

Liquid biopsies, particularly those involving circulating tumor DNA (ctDNA), are rapidly emerging as a non-invasive alternative to tumor biopsies. However, clinical applications of ctDNA analysis in hepatocellular carcinoma (HCC) have not been fully elucidated.

Methods

We measured the amount of plasma-derived cell-free DNA (cfDNA) in HCC patients before (n = 100) and a few days after treatment (n = 87), including radiofrequency ablation, transarterial chemoembolization, and molecular-targeted agents (MTAs), and prospectively analyzed their associations with clinical parameters and prognosis. TERT promoter mutations in cfDNA were analyzed using droplet digital PCR. Furthermore, we performed a comprehensive mutational analysis of post-treatment cfDNA via targeted ultra-deep sequencing (22,000× coverage) in a panel of 275 cancer-related genes in selected patients.

Results

Plasma cfDNA levels increased significantly according to HCC clinical stage, and a high cfDNA level was independently associated with a poor prognosis. TERT promoter mutations were detected in 45% of all cases but were not associated with any clinical characteristics. cfDNA levels increased significantly a few days after treatment, and a greater increase in post-treatment cfDNA levels was associated with a greater therapeutic response to MTAs. The detection rate of TERT mutations increased to 57% using post-treatment cfDNA, suggesting that the ctDNA was enriched. Targeted ultra-deep sequencing using post-treatment cfDNA after administering lenvatinib successfully detected various gene mutations and obtained promising results in lenvatinib-responsive cases.

Conclusions

Post-treatment cfDNA analysis may facilitate the construction of biomarkers for predicting MTA treatment effects.

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Data availability

All data generated or analyzed during this study are included in this published article.

Abbreviations

HCC:

Hepatocellular carcinoma

ctDNA:

Circulating tumor DNA

cfDNA:

Cell-free DNA

RFA:

Radiofrequency ablation

TACE:

Transarterial chemoembolization

MTA:

Molecular-targeted agent

BCLC staging system:

Barcelona clinic liver cancer staging system

AFP:

Alpha-fetoprotein

AFP-L3:

Lens culinaris agglutinin-reactive fraction of AFP

DCP:

Des-gamma-carboxy prothrombin

ddPCR:

Droplet digital PCR

TERT:

Telomerase reverse transcriptase

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Funding

This research was supported by Bristol-Myers Squibb Research Grant, Takeda Science Foundation, MSD Life Science Foundation, The Naito Foundation, Life Science Foundation of Japan, The Cell Science Research Foundation (H.N.), the Charitable Trust Laboratory Medicine Research Foundation of Japan (M.S.), JSPS KAKENHI Grant Numbers JP18K15741 (T.N.), JP18H02789 (H.N.), and JP20K08352 (R.T.), and the Research Program on Hepatitis from Japan Agency for Medical Research and Development (AMED) under Grant Number JP18fk0210040, JP19fk0210040, JP20fk0210040 (H.N. and K.K.), JP19fk0210059, and JP20fk0210059 (H.N.).

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Authors and Affiliations

  1. Department of Gastroenterology, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8655, Japan

    Takuma Nakatsuka, Hayato Nakagawa, Yuki Hayata, Taijiro Wake, Tomoharu Yamada, Mizuki Nishibatake Kinoshita, Ryo Nakagomi, Masaya Sato, Tatsuya Minami, Koji Uchino, Kenichiro Enooku, Yotaro Kudo, Yasuo Tanaka, Takahiro Kishikawa, Motoyuki Otsuka, Ryosuke Tateishi & Kazuhiko Koike

  2. Department of Clinical Laboratory Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8655, Japan

    Masaya Sato

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  1. Takuma Nakatsuka
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Contributions

TN, HN: conceptualization; TN, HN: data curation; TN, HN: formal analysis; YH, TW, TY, MNK, RN, MS, TM, KU, KE, YK, YT: material support; TK, MO: technical support; TN: drafting of the manuscript; HN, RT: critical review of the manuscript; RT, KK: study supervision.

Corresponding author

Correspondence to Hayato Nakagawa.

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The authors declare no conflict of interest.

Ethics approval

The study protocol was approved by the University of Tokyo Medical Research Center Ethics Committee (Approval Number: 11839).

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All patients provided written informed consent.

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Nakatsuka, T., Nakagawa, H., Hayata, Y. et al. Post-treatment cell-free DNA as a predictive biomarker in molecular-targeted therapy of hepatocellular carcinoma. J Gastroenterol 56, 456–469 (2021). https://doi.org/10.1007/s00535-021-01773-4

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