Detection of mutations in EGFR in circulating lung-cancer cells.
Detection of mutations in EGFR in circulating lung-cancer cells.
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DOI：
10.1056/nejmoa0800668
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发表时间：
2008-07-24
期刊：
影响因子：
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通讯作者：
Haber DA
中科院分区：
文献类型：
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作者：
Maheswaran S;Sequist LV;Nagrath S;Ulkus L;Brannigan B;Collura CV;Inserra E;Diederichs S;Iafrate AJ;Bell DW;Digumarthy S;Muzikansky A;Irimia D;Settleman J;Tompkins RG;Lynch TJ;Toner M;Haber DA
在非小细胞肺癌患者中使用酪氨酸激酶抑制剂靶向表皮生长因子受体基因（EGFR）是有效的，但受到耐药突变的限制。循环肿瘤细胞的分子特征可能为治疗期间肿瘤基因型的无创连续监测提供一种策略。我们使用微流控装置从非小细胞肺癌患者的血液中捕获高度纯化的循环肿瘤细胞，该装置含有涂有抗上皮细胞抗体的微柱。我们使用等位基因特异性聚合酶链反应扩增技术对循环肿瘤细胞中回收的DNA进行了EGFR突变分析，并将结果与同时分离的游离血浆DNA和原始肿瘤活检标本的结果进行了比较。我们从27例转移性非小细胞肺癌患者中分离出循环肿瘤细胞（中位数为每毫升74个细胞）。我们在12名患者中的11名（92%）和12名患者中的4名（33%）匹配的游离血浆DNA中发现了预期的EGFR激活突变（P = 0.009）。我们在接受酪氨酸激酶抑制剂治疗的EGFR突变患者收集的循环肿瘤细胞中检测到T790M突变，该突变赋予耐药性。当预处理肿瘤活检标本中检测到T790M时，突变的存在与无进展生存期降低相关（7.7个月对16.5个月，P<0.001）。循环肿瘤细胞的系列分析表明，捕获细胞数量的减少与放射学肿瘤反应有关；细胞数量的增加与肿瘤进展有关，在某些情况下会出现额外的EGFR突变。对肺癌患者血液中循环肿瘤细胞的分子分析为监测治疗过程中上皮肿瘤基因型的变化提供了可能。
The use of tyrosine kinase inhibitors to target the epidermal growth factor receptor gene (EGFR) in patients with non–small-cell lung cancer is effective but limited by the emergence of drug-resistance mutations. Molecular characterization of circulating tumor cells may provide a strategy for noninvasive serial monitoring of tumor genotypes during treatment. We captured highly purified circulating tumor cells from the blood of patients with non–small-cell lung cancer using a microfluidic device containing microposts coated with antibodies against epithelial cells. We performed EGFR mutational analysis on DNA recovered from circulating tumor cells using allele-specific polymerase-chain-reaction amplification and compared the results with those from concurrently isolated free plasma DNA and from the original tumor-biopsy specimens. We isolated circulating tumor cells from 27 patients with metastatic non–small-cell lung cancer (median number, 74 cells per milliliter). We identified the expected EGFR activating mutation in circulating tumor cells from 11 of 12 patients (92%) and in matched free plasma DNA from 4 of 12 patients (33%) (P = 0.009). We detected the T790M mutation, which confers drug resistance, in circulating tumor cells collected from patients with EGFR mutations who had received tyrosine kinase inhibitors. When T790M was detectable in pretreatment tumor-biopsy specimens, the presence of the mutation correlated with reduced progression-free survival (7.7 months vs. 16.5 months, P<0.001). Serial analysis of circulating tumor cells showed that a reduction in the number of captured cells was associated with a radiographic tumor response; an increase in the number of cells was associated with tumor progression, with the emergence of additional EGFR mutations in some cases. Molecular analysis of circulating tumor cells from the blood of patients with lung cancer offers the possibility of monitoring changes in epithelial tumor genotypes during the course of treatment.