Intra- and inter-tumor heterogeneity of BRAF(V600E))mutations in primary and metastatic melanoma.
Intra- and inter-tumor heterogeneity of BRAF(V600E))mutations in primary and metastatic melanoma.
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DOI：
10.1371/journal.pone.0029336
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Polsky D
中科院分区：
文献类型：
--
作者：
Yancovitz M;Litterman A;Yoon J;Ng E;Shapiro RL;Berman RS;Pavlick AC;Darvishian F;Christos P;Mazumdar M;Osman I;Polsky D
使用致癌蛋白的小分子抑制剂作为癌症疗法的基本原理至少部分地取决于转移性肿瘤相对于突变致癌基因主要是克隆的假设。随着BRAFV 600 E作为治疗靶点的出现，我们使用BRAFV 600 E突变作为克隆性标志物的检测来研究黑色素瘤中的肿瘤内和肿瘤间异质性。对来自73名患者的112个肿瘤进行BRAF突变体特异性PCR（MS-PCR）和常规测序，包括具有匹配的原发性和转移性标本的患者（n = 18）。 19例患者的组织来自多个转移部位。使用常规测序在36/112（32%）黑素瘤中检测到突变，使用MS-PCR在85/112（76%）黑素瘤中检测到突变。MS-PCR检测突变BRAFV 600 E等位基因的更好的灵敏度不是由于存在污染的正常组织，这表明肿瘤由不同BRAF基因型的亚克隆组成。为了确定肿瘤亚克隆是否存在于单个原发性黑色素瘤中，我们在9例病例中进行了激光显微切割和突变检测，通过测序和BRAFV 600 E特异性SNaPshot分析。这些病例中有6例在单个肿瘤内不同的显微切割区域显示了不同比例的BRAFV 600 E和BRAF野生型细胞。使用高灵敏度MS-PCR对来自个体患者的多个转移性样本进行的额外分析显示，5/19（26%）例患者的转移与BRAFV 600 E突变不一致。总之，我们使用了高度敏感的BRAF突变检测方法，并观察到个体黑色素瘤肿瘤标本和个体患者多个标本中BRAFV 600 E突变异质性的实质性证据。考虑到患者对BRAF抑制剂治疗的不同临床反应，这些数据表明，确定临床结局与肿瘤内和肿瘤间异质性之间可能相关性的额外研究可能会取得丰硕成果。
The rationale for using small molecule inhibitors of oncogenic proteins as cancer therapies depends, at least in part, on the assumption that metastatic tumors are primarily clonal with respect to mutant oncogene. With the emergence of BRAFV600E as a therapeutic target, we investigated intra- and inter-tumor heterogeneity in melanoma using detection of the BRAFV600E mutation as a marker of clonality. BRAF mutant-specific PCR (MS-PCR) and conventional sequencing were performed on 112 tumors from 73 patients, including patients with matched primary and metastatic specimens (n = 18). Nineteen patients had tissues available from multiple metastatic sites. Mutations were detected in 36/112 (32%) melanomas using conventional sequencing, and 85/112 (76%) using MS-PCR. The better sensitivity of the MS-PCR to detect the mutant BRAFV600E allele was not due to the presence of contaminating normal tissue, suggesting that the tumor was comprised of subclones of differing BRAF genotypes. To determine if tumor subclones were present in individual primary melanomas, we performed laser microdissection and mutation detection via sequencing and BRAFV600E-specific SNaPshot analysis in 9 cases. Six of these cases demonstrated differing proportions of BRAFV600Eand BRAFwild-type cells in distinct microdissected regions within individual tumors. Additional analyses of multiple metastatic samples from individual patients using the highly sensitive MS-PCR without microdissection revealed that 5/19 (26%) patients had metastases that were discordant for the BRAFV600E mutation. In conclusion, we used highly sensitive BRAF mutation detection methods and observed substantial evidence for heterogeneity of the BRAFV600E mutation within individual melanoma tumor specimens, and among multiple specimens from individual patients. Given the varied clinical responses of patients to BRAF inhibitor therapy, these data suggest that additional studies to determine possible associations between clinical outcomes and intra- and inter-tumor heterogeneity could prove fruitful.
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影响因子：
64.5
作者：
Heidorn SJ;Milagre C;Whittaker S;Nourry A;Niculescu-Duvas I;Dhomen N;Hussain J;Reis-Filho JS;Springer CJ;Pritchard C;Marais R
通讯作者：
Marais R
影响因子：
8.8
作者：
通讯作者：
--
影响因子：
8
作者：
Cooke, S. L.;Ng, C. K. Y.;Melnyk, N.;Garcia, M. J.;Hardcastle, T.;Temple, J.;Langdon, S.;Huntsman, D.;Brenton, J. D.
通讯作者：
Brenton, J. D.
影响因子：
7.5
作者：
Dias-Santagata, Dora;Quynh Lam;Hoang, Mai P.
通讯作者：
Hoang, Mai P.
影响因子：
10.3
作者：
Lin, Jingrong;Takata, Minoru;Saida, Toshiaki
通讯作者：
Saida, Toshiaki