A High-Throughput Panel for Identifying Clinically Relevant Mutation Profiles in Melanoma
A High-Throughput Panel for Identifying Clinically Relevant Mutation Profiles in Melanoma
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DOI：
10.1158/1535-7163.mct-11-0676
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发表时间：
2012-04-01
影响因子：
5.7
通讯作者：
Hayward, Nicholas K.
中科院分区：
文献类型：
--
作者：
Dutton-Regester, Ken;Irwin, Darryl;Hayward, Nicholas K.
基于分子的靶向药物在癌症治疗中的成功引发了个性化治疗领域的广泛研究工作。然而，此类疗法的成功应用取决于患者肿瘤内是否存在可赋予临床疗效或耐药性的突变。基于这些发现，我们开发了一个高通量突变面板，用于识别黑色素瘤中经常发生的和临床相关的突变。对癌症体细胞突变目录数据库进行了广泛的文献检索和查询，发现了 1,000 多种黑色素瘤突变。我们采用过滤策略重点关注适合靶向药物开发的突变，最初使用 Sequenom MassARRAY 系统筛选了 271 个样本中的 120 个已知突变。 17个基因共检测到252个突变，频率最高的是BRAF（n=154，57%）、NRAS（n=55，20%）、CDK4（n=8，3%）、PTK2B（n=7，2.5%）和ERBB4（n=5，2%）。基于这一初步发现筛选，总共设计了 46 项检测，询问 20 个基因的 39 个突变，以开发黑色素瘤特异性组。这些检测采用针对灵敏突变检测而优化的严格检测设计参数，分布在 8 个孔的多重检测中。最终的黑色素瘤特异性突变面板是一种经济高效、灵敏、高通量的方法，用于识别与黑色素瘤分子疗法临床相关的突变。当用于临床研究环境时，该小组可以使用新型或现有的分子靶向药物快速准确地识别潜在有效的治疗策略。摩尔癌症治疗； 11(4)； 888-97。 (C) 2012 年 AACR。
Success with molecular-based targeted drugs in the treatment of cancer has ignited extensive research efforts within the field of personalized therapeutics. However, successful application of such therapies is dependent on the presence or absence of mutations within the patient's tumor that can confer clinical efficacy or drug resistance. Building on these findings, we developed a high-throughput mutation panel for the identification of frequently occurring and clinically relevant mutations in melanoma. An extensive literature search and interrogation of the Catalogue of Somatic Mutations in Cancer database identified more than 1,000 melanoma mutations. Applying a filtering strategy to focus on mutations amenable to the development of targeted drugs, we initially screened 120 known mutations in 271 samples using the Sequenom MassARRAY system. A total of 252 mutations were detected in 17 genes, the highest frequency occurred in BRAF (n = 154, 57%), NRAS (n = 55, 20%), CDK4 (n = 8, 3%), PTK2B (n = 7, 2.5%), and ERBB4 (n = 5, 2%). Based on this initial discovery screen, a total of 46 assays interrogating 39 mutations in 20 genes were designed to develop a melanoma-specific panel. These assays were distributed in multiplexes over 8 wells using strict assay design parameters optimized for sensitive mutation detection. The final melanoma-specific mutation panel is a cost effective, sensitive, high-throughput approach for identifying mutations of clinical relevance to molecular-based therapeutics for the treatment of melanoma. When used in a clinical research setting, the panel may rapidly and accurately identify potentially effective treatment strategies using novel or existing molecularly targeted drugs. Mol Cancer Ther; 11(4); 888-97. (C) 2012 AACR.