Towards Personalized Medicine in Melanoma: Implementation of a Clinical Next-Generation Sequencing Panel.
Towards Personalized Medicine in Melanoma: Implementation of a Clinical Next-Generation Sequencing Panel.
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DOI：
10.1038/s41598-017-00606-w
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发表时间：
2017-03-29
影响因子：
4.6
通讯作者：
Botella Estrada R
中科院分区：
文献类型：
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作者：
de Unamuno Bustos B;Murria Estal R;Pérez Simó G;de Juan Jimenez I;Escutia Muñoz B;Rodríguez Serna M;Alegre de Miquel V;Llavador Ros M;Ballester Sánchez R;Nagore Enguídanos E;Palanca Suela S;Botella Estrada R
由于靶向特定基因突变的新型疗法的发展，在黑色素瘤患者的诊断和管理中越来越多地常规进行分子诊断。下一代测序（NGS）技术的发展使得能够在单个测定中对多个癌症驱动基因进行测序，并且提高了突变检测的灵敏度。本研究的主要目的是设计和实施一个黑色素瘤特异性测序面板，并在一系列原发性黑色素瘤样本中鉴定体细胞突变谱。设计了一个自定义面板，以覆盖35个黑色素瘤相关基因的编码区。样本组平均覆盖率为每个扩增子2，575.5个读数，92.8%的靶碱基覆盖率≥500×。深度覆盖使得能够以低至0.5%的突变等位基因频率灵敏地发现突变。85%（85/100）的黑色素瘤至少有一个体细胞突变。最常见的突变基因是BRAF（50%;50/199）、NRAS（15%;15/100）、PREX 2（14%;14/100）、GRIN 2A（13%;13/100）和ERBB 4（12%;12/100）。与传统的分子方法相比，基于NGS的分析的周转时间和成本降低。本研究的结果证明了定制设计的目标NGS面板的成本效益和可行性，并建议将目标NGS实施到日常实践中。
Molecular diagnostics are increasingly performed routinely in the diagnosis and management of patients with melanoma due to the development of novel therapies that target specific genetic mutations. The development of next-generation sequencing (NGS) technologies has enabled to sequence multiple cancer-driving genes in a single assay, with improved sensitivity in mutation detection. The main objective of this study was the design and implementation of a melanoma-specific sequencing panel, and the identification of the spectrum of somatic mutations in a series of primary melanoma samples. A custom panel was designed to cover the coding regions of 35 melanoma-related genes. Panel average coverage was 2,575.5 reads per amplicon, with 92,8% of targeted bases covered ≥500×. Deep coverage enabled sensitive discovery of mutations in as low as 0.5% mutant allele frequency. Eighty-five percent (85/100) of the melanomas had at least one somatic mutation. The most prevalent mutated genes were BRAF (50%;50/199), NRAS (15%;15/100), PREX2 (14%;14/100), GRIN2A (13%;13/100), and ERBB4 (12%;12/100). Turn-around-time and costs for NGS-based analysis was reduced in comparison to conventional molecular approaches. The results of this study demonstrate the cost-effectiveness and feasibility of a custom-designed targeted NGS panel, and suggest the implementation of targeted NGS into daily routine practice.