Zacher Angela, Kaulich Kerstin, Stepanow Stefanie, Wolter Marietta, Köhrer Karl, Felsberg Jörg, Malzkorn Bastian, Reifenberger Guido
Department of Neuropathology, Heinrich Heine University Düsseldorf, Düsseldorf, Germany.
German Cancer Consortium (DKTK), German Cancer Research Center (DKFZ) Heidelberg, partner site Essen/Düsseldorf, Germany.
Brain Pathol. 2017 Mar;27(2):146-159. doi: 10.1111/bpa.12367. Epub 2016 Apr 19.
Current classification of gliomas is based on histological criteria according to the World Health Organization (WHO) classification of tumors of the central nervous system. Over the past years, characteristic genetic profiles have been identified in various glioma types. These can refine tumor diagnostics and provide important prognostic and predictive information. We report on the establishment and validation of gene panel next generation sequencing (NGS) for the molecular diagnostics of gliomas. We designed a glioma-tailored gene panel covering 660 amplicons derived from 20 genes frequently aberrant in different glioma types. Sensitivity and specificity of glioma gene panel NGS for detection of DNA sequence variants and copy number changes were validated by single gene analyses. NGS-based mutation detection was optimized for application on formalin-fixed paraffin-embedded tissue specimens including small stereotactic biopsy samples. NGS data obtained in a retrospective analysis of 121 gliomas allowed for their molecular classification into distinct biological groups, including (i) isocitrate dehydrogenase gene (IDH) 1 or 2 mutant astrocytic gliomas with frequent α-thalassemia/mental retardation syndrome X-linked (ATRX) and tumor protein p53 (TP53) gene mutations, (ii) IDH mutant oligodendroglial tumors with 1p/19q codeletion, telomerase reverse transcriptase (TERT) promoter mutation and frequent Drosophila homolog of capicua (CIC) gene mutation, as well as (iii) IDH wildtype glioblastomas with frequent TERT promoter mutation, phosphatase and tensin homolog (PTEN) mutation and/or epidermal growth factor receptor (EGFR) amplification. Oligoastrocytic gliomas were genetically assigned to either of these groups. Our findings implicate gene panel NGS as a promising diagnostic technique that may facilitate integrated histological and molecular glioma classification.
目前，胶质瘤的分类是基于世界卫生组织（WHO）中枢神经系统肿瘤分类的组织学标准。在过去几年中，已在各种类型的胶质瘤中鉴定出特征性的基因谱。这些基因谱可以完善肿瘤诊断，并提供重要的预后和预测信息。我们报告了用于胶质瘤分子诊断的基因panel二代测序（NGS）技术的建立和验证情况。我们设计了一个针对胶质瘤的基因panel，涵盖了来自20个在不同类型胶质瘤中经常发生异常的基因的660个扩增子。通过单基因分析验证了胶质瘤基因panel NGS检测DNA序列变异和拷贝数变化的敏感性和特异性。基于NGS的突变检测针对福尔马林固定石蜡包埋组织标本（包括小的立体定向活检样本）进行了优化。对121例胶质瘤的回顾性分析中获得的NGS数据，使其能够被分子分类为不同的生物学组，包括：（i）异柠檬酸脱氢酶基因（IDH）1或2突变的星形细胞胶质瘤，伴有频繁的X连锁α地中海贫血/智力发育迟缓综合征（ATRX）和肿瘤蛋白p53（TP53）基因突变；（ii）IDH突变的少突胶质细胞瘤，伴有1p/19q共缺失、端粒酶逆转录酶（TERT）启动子突变和频繁的果蝇同源基因capicua（CIC）基因突变；以及（iii）IDH野生型胶质母细胞瘤，伴有频繁的TERT启动子突变、磷酸酶和张力蛋白同源物（PTEN）突变和/或表皮生长因子受体（EGFR）扩增。少突星形细胞瘤在基因上被归为这些组中的一类。我们的研究结果表明，基因panel NGS是一种有前景的诊断技术，可能有助于整合组织学和分子胶质瘤分类。
