Post-treatment cell-free DNA as a predictive biomarker in molecular-targeted therapy of hepatocellular carcinoma
Post-treatment cell-free DNA as a predictive biomarker in molecular-targeted therapy of hepatocellular carcinoma
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治疗后无细胞DNA作为肝细胞癌分子靶向治疗的预测性生物标志物
DOI：
10.1007/s00535-021-01773-4
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发表时间：
2021-03-12
影响因子：
6.3
通讯作者：
Koike, Kazuhiko
中科院分区：
文献类型：
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作者：
Nakatsuka, Takuma;Nakagawa, Hayato;Koike, Kazuhiko
背景液体活检，尤其是涉及循环肿瘤DNA(CtDNA)的活检，正迅速成为肿瘤活检的一种非侵入性替代方法。然而，ctDNA分析在肝细胞癌中的临床应用尚未完全阐明。方法检测100例肝癌患者治疗前(n=100)和治疗后几天(n=87)患者血浆中游离DNA(CfDNA)的含量，并前瞻性分析其与临床参数和预后的关系。用Droplet数字聚合酶链式反应分析cfDNA中TERT启动子的突变。此外，我们通过对选定患者的275个癌症相关基因进行定向超深度测序(覆盖范围22,000倍)，对治疗后的cfDNA进行了全面的突变分析。结果随着肝细胞癌临床分期的不同，血浆cfDNA水平显著升高，cfDNA水平高与预后不良独立相关。45%的病例检测到TERT启动子突变，但与任何临床特征无关。CfDNA水平在治疗后几天显著升高，治疗后cfDNA水平的更大增加与MTA的治疗反应更大相关。经cfDNA处理后，TERT突变的检出率提高到57%，提示ctDNA被富集化。在给予lenvatinib后，使用后处理cfDNA进行靶向超深测序，成功地检测到各种基因突变，并在lenvatinib敏感病例中获得了令人振奋的结果。结论治疗后cfDNA分析有助于构建预测MTA治疗效果的生物标志物。
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 (22,000x 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.