Circulating Tumor DNA Analysis for Liver Cancers and Its Usefulness as a Liquid Biopsy.
Circulating Tumor DNA Analysis for Liver Cancers and Its Usefulness as a Liquid Biopsy.
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DOI：
10.1016/j.jcmgh.2015.06.009
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发表时间：
2015-09
影响因子：
7.2
通讯作者：
Chayama K
中科院分区：
文献类型：
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作者：
Ono A;Fujimoto A;Yamamoto Y;Akamatsu S;Hiraga N;Imamura M;Kawaoka T;Tsuge M;Abe H;Hayes CN;Miki D;Furuta M;Tsunoda T;Miyano S;Kubo M;Aikata H;Ochi H;Kawakami YI;Arihiro K;Ohdan H;Nakagawa H;Chayama K
在血液的无细胞部分中发现了携带肿瘤特异性序列改变的循环肿瘤DNA（ctDNA）。肝癌肿瘤标本很难获得，需要非侵入性方法来评估癌症进展和表征潜在的基因组特征。我们分析了46例接受肝切除术或肝移植的肝细胞癌患者，他们的全基因组测序数据可用。我们设计了针对每个肿瘤的体细胞重排的个性化测定来定量血清ctDNA。使用来自经导管动脉化疗栓塞（TACE）后复发性肝癌患者的无细胞DNA配对原发性肿瘤组织DNA进行外显子组测序。我们在46名患者中的7名患者手术前成功检测到100 μL血清样本中的ctDNA，并随着疾病进展而增加。ctDNA阳性组的累积复发率和肝外转移率在统计学上显著低于ctDNA阴性组（分别为P = 0.0102和0.0386）。多变量分析确定ctDNA（OR 6.10; 95%CI，1.11-33.33，P = 0.038）是门静脉（VP）显微血管浸润的独立预测因子。我们通过外显子组测序在TACE后的无细胞DNA中鉴定出45个非同义体细胞突变，在原发性肿瘤组织中鉴定出71个非同义体细胞突变。我们在两个样本中鉴定了25个常见突变，原发性肿瘤中鉴定的83%的突变可以在无细胞DNA中检测到。ctDNA的存在反映了肿瘤的进展，ctDNA的检测可以预测VP和复发，特别是2年内的肝外转移。我们的研究证明了ctDNA检测和游离DNA测序分析对肝癌个性化治疗的有用性。
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. 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). 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. 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.