Department of Medical Oncology, Princess Margaret Hospital, University Health Network, University of Toronto, Toronto, Ontario, Canada.
Int J Cancer. 2013 Apr 1;132(7):1547-55. doi: 10.1002/ijc.27817. Epub 2012 Oct 11.
The successes of targeted drugs with companion predictive biomarkers and the technological advances in gene sequencing have generated enthusiasm for evaluating personalized cancer medicine strategies using genomic profiling. We assessed the feasibility of incorporating real-time analysis of somatic mutations within exons of 19 genes into patient management. Blood, tumor biopsy and archived tumor samples were collected from 50 patients recruited from four cancer centers. Samples were analyzed using three technologies: targeted exon sequencing using Pacific Biosciences PacBio RS, multiplex somatic mutation genotyping using Sequenom MassARRAY and Sanger sequencing. An expert panel reviewed results prior to reporting to clinicians. A clinical laboratory verified actionable mutations. Fifty patients were recruited. Nineteen actionable mutations were identified in 16 (32%) patients. Across technologies, results were in agreement in 100% of biopsy specimens and 95% of archival specimens. Profiling results from paired archival/biopsy specimens were concordant in 30/34 (88%) patients. We demonstrated that the use of next generation sequencing for real-time genomic profiling in advanced cancer patients is feasible. Additionally, actionable mutations identified in this study were relatively stable between archival and biopsy samples, implying that cancer mutations that are good predictors of drug response may remain constant across clinical stages.
靶向药物与伴随预测性生物标志物的成功以及基因测序技术的进步，激发了人们对利用基因组分析评估个性化癌症药物策略的热情。我们评估了将 19 个基因外显子中的体细胞突变实时分析纳入患者管理的可行性。从四个癌症中心招募的 50 名患者采集了血液、肿瘤活检和存档的肿瘤样本。使用三种技术对样本进行了分析：使用 Pacific Biosciences PacBio RS 进行靶向外显子测序、使用 Sequenom MassARRAY 和 Sanger 测序进行多重体细胞突变基因分型。专家小组在向临床医生报告前对结果进行了审查。临床实验室验证了可采取的突变。共招募了 50 名患者。在 16 名（32%）患者中发现了 19 个可采取的突变。在所有技术中，活检标本的结果一致性为 100%，存档标本的结果一致性为 95%。34 对存档/活检标本的分析结果一致，占 30/34（88%）患者。我们证明了在晚期癌症患者中使用下一代测序进行实时基因组分析是可行的。此外，本研究中鉴定的可采取的突变在存档和活检样本之间相对稳定，这意味着对药物反应有良好预测性的癌症突变可能在整个临床阶段保持不变。
