Department of Pediatrics, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA.
Clin Cancer Res. 2012 Feb 1;18(3):748-57. doi: 10.1158/1078-0432.CCR-11-2056. Epub 2011 Dec 5.
In contrast to the numerous broad screens for oncogene mutations in adult cancers, few such screens have been conducted in pediatric solid tumors. To identify novel mutations and potential therapeutic targets in pediatric cancers, we conducted a high-throughput Sequenom-based analysis in large sets of several major pediatric solid cancers, including neuroblastoma, Ewing sarcoma, rhabdomyosarcoma (RMS), and desmoplastic small round cell tumor (DSRCT).
We designed a highly multiplexed Sequenom-based assay to interrogate 275 recurrent mutations across 29 genes. Genomic DNA was extracted from 192 neuroblastoma, 75 Ewing sarcoma, 89 RMS, and 24 DSRCT samples. All mutations were verified by Sanger sequencing.
Mutations were identified in 13% of neuroblastoma samples, 4% of Ewing sarcoma samples, 21.1% of RMS samples, and no DSRCT samples. ALK mutations were present in 10.4% of neuroblastoma samples. The remainder of neuroblastoma mutations involved the BRAF, RAS, and MAP2K1 genes and were absent in samples harboring ALK mutations. Mutations were more common in embryonal RMS (ERMS) samples (28.3%) than alveolar RMS (3.5%). In addition to previously identified RAS and FGFR4 mutations, we report for the first time PIK3CA and CTNNB1 (β-catenin) mutations in 5% and 3.3% of ERMS, respectively.
In ERMS, Ewing sarcoma, and neuroblastoma, we identified novel occurrences of several oncogene mutations recognized as drivers in other cancers. Overall, neuroblastoma and ERMS contain significant subsets of cases with nonoverlapping mutated genes in growth signaling pathways. Tumor profiling can identify a subset of pediatric solid tumor patients as candidates for kinase inhibitors or RAS-targeted therapies.
与大量针对成人癌症中致癌基因突变的广谱筛查相比，针对儿科实体瘤的此类筛查较少。为了在儿科癌症中发现新的突变和潜在的治疗靶点，我们对包括神经母细胞瘤、尤文肉瘤、横纹肌肉瘤（RMS）和促结缔组织增生小圆细胞肿瘤（DSRCT）在内的几种主要儿科实体瘤的大型样本进行了高通量基于Sequenom 的分析。
我们设计了一种高度多重化的基于Sequenom 的检测方法，以检测 29 个基因中的 275 个复发性突变。从 192 例神经母细胞瘤、75 例尤文肉瘤、89 例 RMS 和 24 例 DSRCT 样本中提取基因组 DNA。所有突变均通过 Sanger 测序验证。
在 13%的神经母细胞瘤样本、4%的尤文肉瘤样本、21.1%的 RMS 样本和没有 DSRCT 样本中发现了突变。ALK 突变存在于 10.4%的神经母细胞瘤样本中。其余的神经母细胞瘤突变涉及 BRAF、RAS 和 MAP2K1 基因，并且不存在 ALK 突变的样本中。在胚胎性 RMS（ERMS）样本中，突变更为常见（28.3%），而在肺泡 RMS（3.5%）中则较少。除了先前鉴定的 RAS 和 FGFR4 突变外，我们还首次在 5%的 ERMS 和 3.3%的 RMS 中报告了 PIK3CA 和 CTNNB1（β-连环蛋白）突变。
在 ERMS、尤文肉瘤和神经母细胞瘤中，我们发现了几种致癌基因突变的新发生，这些突变被认为是其他癌症的驱动因素。总体而言，神经母细胞瘤和 ERMS 包含了大量具有非重叠生长信号通路突变基因的病例。肿瘤分析可以确定一部分儿科实体瘤患者作为激酶抑制剂或 RAS 靶向治疗的候选者。
