Complex Tumor Genomes Inferred from Single Circulating Tumor Cells by Array-CGH and Next-Generation Sequencing
Complex Tumor Genomes Inferred from Single Circulating Tumor Cells by Array-CGH and Next-Generation Sequencing
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DOI：
10.1158/0008-5472.can-12-4140
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发表时间：
2013-05-15
期刊：
影响因子：
11.2
通讯作者：
Speicher, Michael R.
中科院分区：
文献类型：
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作者：
Heitzer, Ellen;Auer, Martina;Speicher, Michael R.
从原发性癌症和转移瘤释放到血液中的循环肿瘤细胞（CTC）反映了肿瘤基因型的现状，这些基因型容易发生变化。在这里，我们使用阵列比较基因组杂交（CGH）和下一代测序进行了第一次全面的CTC基因组分析。我们使用了U。S.食品和药物管理局批准的CellSearch系统，该系统在37例IV期结直肠癌患者中的21例（范围，1-202/7.5 mL样品）中检测到CTC。总的来说，我们能够从6名患者中分离出37个完整的CTC，并在这些多个结直肠癌相关的拷贝数变化中进行鉴定，其中许多也存在于相应的原发性肿瘤中。然后，我们对一组68个结直肠癌相关基因进行了大规模平行测序，以比较其中两名患者的原发性肿瘤，转移灶和相应CTC的突变谱。已知驱动基因的突变[ e]。例如，在一个实施例中，在原发性肿瘤中发现的大肠腺瘤性息肉病（APC）、KRAS或PIK 3CA]和在相应的CTC中也检测到转移。然而，我们也观察到仅在CTC中的突变。为了确定这些突变是否来源于原发性肿瘤中的小亚克隆或代表转移性细胞的新变体，我们对原发性肿瘤和转移进行了额外的深度测序，并应用定制的统计算法进行分析。我们发现，最初仅在CTC中发现的大多数突变在同一患者的原发肿瘤和转移瘤中也以亚克隆水平存在。这项研究为在癌症患者中使用CTC作为液体活检铺平了道路，为监测在进展，治疗和复发过程中容易发生变化的肿瘤基因组提供了更有效的选择。Cancer Res; 73（10）; 2965-75. (C)2013年AACR。
Circulating tumor cells (CTC) released into blood from primary cancers and metastases reflect the current status of tumor genotypes, which are prone to changes. Here, we conducted the first comprehensive genomic profiling of CTCs using array-comparative genomic hybridization (CGH) and next-generation sequencing. We used the U. S. Food and Drug Administration-cleared CellSearch system, which detected CTCs in 21 of 37 patients (range, 1-202/7.5 mL sample) with stage IV colorectal carcinoma. In total, we were able to isolate 37 intact CTCs from six patients and identified in those multiple colorectal cancer-associated copy number changes, many of which were also present in the respective primary tumor. We then used massive parallel sequencing of a panel of 68 colorectal cancer-associated genes to compare the mutation spectrum in the primary tumors, metastases, and the corresponding CTCs from two of these patients. Mutations in known driver genes [ e. g., adenomatous polyposis coli (APC), KRAS, or PIK3CA] found in the primary tumor and metastasis were also detected in corresponding CTCs. However, we also observed mutations exclusively in CTCs. To address whether these mutations were derived from a small subclone in the primary tumor or represented new variants of metastatic cells, we conducted additional deep sequencing of the primary tumor and metastasis and applied a customized statistical algorithm for analysis. We found that most mutations initially found only in CTCs were also present at subclonal level in the primary tumors andmetastases from the same patient. This study paves the way to use CTCs as a liquid biopsy in patients with cancer, providing more effective options to monitor tumor genomes that are prone to change during progression, treatment, and relapse. Cancer Res; 73(10); 2965-75. (C) 2013 AACR.