Endris Volker, Stenzinger Albrecht, Pfarr Nicole, Penzel Roland, Möbs Markus, Lenze Dido, Darb-Esfahani Silvia, Hummel Michael, Jung Andreas, Lehmann Ulrich, Kreipe Hans, Kirchner Thomas, Büttner Reinhard, Jochum Wolfram, Höfler Gerald, Dietel Manfred, Weichert Wilko, Schirmacher Peter
Institute of Pathology, University Hospital Heidelberg, Im Neuenheimer Feld, 224, Heidelberg, Germany.
Institute of Pathology, Technical University of Munich (TUM), Munich, Germany.
Virchows Arch. 2016 Jun;468(6):697-705. doi: 10.1007/s00428-016-1919-8. Epub 2016 Mar 22.
With the approval of olaparib as monotherapy treatment in platinum-sensitive, relapsed high-grade serous ovarian cancer by the European Medical Agency (EMA), comprehensive genotyping of BRCA1 and BRCA2 in tumor tissue has become a mandatory pre-therapeutic test. This requires significant advances in routine tumor test methodologies due to the large size of both genes and the lack of mutational hot spots. Classical focused screening approaches, like Sanger sequencing, do not allow for a sensitive, rapid, and economic analysis of tumor tissue. Next-generation sequencing (NGS) approaches employing targeted panels for BRCA1/2 to interrogate formalin-fixed and paraffin-embedded tumor samples from either surgical resection or biopsy specimens can overcome these limitations. Although focused NGS methods have been implemented by few centers in routine molecular diagnostics for the analysis of some druggable oncogenic mutations, the reliable diagnostic testing of the entire coding regions of BRCA1 and BRCA2 was a new challenge requiring extensive technological improvement and quality management. Here, we describe the implementation and results of the first round robin trial for BRCA1/2 mutation testing in tumor tissue that was conducted in central Europe on May 2015, shortly after the approval and prior to the official release of olaparib. The high success rate of 81 % (21/26 test centers) demonstrates that BRCA1/2 multicenter mutation testing is well feasible in FFPE tumor tissue, extending to other tumor entities beyond ovarian cancer. The high number of test centers passing the trial demonstrates the success of the concerted efforts by German, Swiss, and Austrian pathology centers to ensure quality-controlled NGS-based testing and proves the potential of this technology in routine molecular pathology. On the basis of our results, we provide recommendations for predictive testing of tumor tissue for BRCA1/2 to clinical decision making in ovarian cancer patients.
随着欧洲药品管理局（EMA）批准奥拉帕利作为铂敏感、复发高级别浆液性卵巢癌的单药治疗药物，肿瘤组织中BRCA1和BRCA2的全面基因分型已成为强制性的治疗前检测。由于这两个基因的规模较大且缺乏突变热点，这就要求常规肿瘤检测方法有重大进展。传统的聚焦筛查方法，如桑格测序，无法对肿瘤组织进行灵敏、快速且经济的分析。采用针对BRCA1/2的靶向panel来检测手术切除或活检标本中福尔马林固定、石蜡包埋肿瘤样本的新一代测序（NGS）方法可以克服这些局限性。尽管少数中心已在常规分子诊断中采用聚焦NGS方法来分析一些可靶向致癌突变，但对BRCA1和BRCA2整个编码区进行可靠的诊断检测仍是一项新挑战，需要广泛的技术改进和质量管理。在此，我们描述了2015年5月在中欧进行的肿瘤组织中BRCA1/2突变检测首轮轮值试验的实施情况和结果，该试验在奥拉帕利获批后不久且正式发布前进行。81%（21/26个检测中心）的高成功率表明，BRCA1/2多中心突变检测在FFPE肿瘤组织中切实可行，并且可扩展到卵巢癌以外的其他肿瘤实体。大量检测中心通过该试验证明了德国、瑞士和奥地利病理中心为确保基于NGS的质量控制检测而共同努力的成功，并证明了该技术在常规分子病理学中的潜力。基于我们的结果，我们为卵巢癌患者肿瘤组织BRCA1/2预测检测到临床决策提供了建议。
