Memorial Sloan Kettering-Integrated Mutation Profiling of Actionable Cancer Targets (MSK-IMPACT) A Hybridization Capture-Based Next-Generation Sequencing Clinical Assay for Solid Tumor Molecular Oncology
Memorial Sloan Kettering-Integrated Mutation Profiling of Actionable Cancer Targets (MSK-IMPACT) A Hybridization Capture-Based Next-Generation Sequencing Clinical Assay for Solid Tumor Molecular Oncology
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DOI：
10.1016/j.jmoldx.2014.12.006
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发表时间：
2015-05-01
影响因子：
4.1
通讯作者：
Berger, Michael F.
中科院分区：
文献类型：
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作者：
Cheng, Donavan T.;Mitchell, Talia N.;Berger, Michael F.
特定遗传改变作为关键致癌驱动因素的鉴定和靶向治疗的开发共同改变了临床肿瘤学，并迫切需要增加临床基因分型的广度和通量。下一代测序检测允许有效和无偏倚地检测临床上可操作的突变。为了实现实体瘤患者的精确肿瘤学，我们开发了Memorial Sloan Kettering-可操作癌症靶点的集成突变分析（MSK-IMPACT），这是一种基于杂交捕获的下一代测序分析，用于所有外显子和选定内含子的靶向深度测序。福尔马林固定石蜡包埋肿瘤中的341个关键癌症基因。捕获来自患者匹配的肿瘤和正常样品的条形码文库，测序，并进行定制分析管道以鉴定体细胞突变。通过广泛的分析验证，MSK-IMPACT的灵敏度，特异性，重现性进行了评估。我们测试了284个肿瘤样本，这些样本在19个癌症基因的47个外显子中具有先前已知的点突变和插入/缺失。所有已知的变异体都被准确地检测到，并且运行间和运行内重复的重现性很高。热点突变的低频变异的检测限约为2%，非热点突变的检测限约为5%。拷贝数改变和结构重排也被可靠地检测到。MSK-IMPACT在临床实体瘤样本中具有高准确性和灵敏度的致癌DNA改变。肿瘤和患者匹配的正常样本的配对分析能够明确检测体细胞突变，以指导治疗决策。
The identification of specific genetic alterations as key oncogenic drivers and the development of targeted therapies are together transforming clinical oncology and creating a pressing need for increased breadth and throughput of clinical genotyping. Next-generation sequencing assays allow the efficient and unbiased detection of clinically actionable mutations. To enable precision oncology in patients with solid tumors, we developed Memorial Sloan Kettering-Integrated Mutation Profiling of Actionable Cancer Targets (MSK-IMPACT), a hybridization capture-based next-generation sequencing assay for targeted deep sequencing of all exons and selected introns. of 341 key cancer genes in formalin-fixed, paraffin-embedded tumors. Barcoded Libraries from patient-matched tumor and normal samples were captured, sequenced, and subjected to a custom analysis pipeline to identify somatic mutations. Sensitivity, specificity, reproducibility of MSK-IMPACT were assessed through extensive analytical validation. We tested 284 tumor samples with previously known point mutations and insertions/deletions in 47 exons of 19 cancer genes. All known variants were accurately detected, and there was high reproducibility of inter- and intrarun replicates. The detection Limit for Low-frequency variants was approximately 2% for hotspot mutations and 5% for nonhotspot mutations. Copy number alterations and structural rearrangements were also reliably detected. MSK-IMPACT profiles oncogenic DNA alterations in clinical solid tumor samples with high accuracy and sensitivity. Paired analysis of tumors and patient-matched normal samples enables unambiguous detection of somatic mutations to guide treatment decisions.