Pathology Department, Hospital del Mar, Barcelona, Spain; Cancer Research Program, Hospital del Mar Medical Research Institute, Barcelona, Spain.
BIOPAT Biopatologia Molecular, Grup Assistència, Barcelona, Spain.
Clin Lung Cancer. 2019 Jul;20(4):e421-e429. doi: 10.1016/j.cllc.2019.02.008. Epub 2019 Feb 26.
Detection of ALK and ROS1 gene rearrangements in non-small-cell lung cancer is required for directing patient care. Although fluorescence in situ hybridization (FISH) and immunohistochemistry have been established as gold standard methods, next-generation sequencing (NGS) platforms are called to be at least equally successful. Comparison of these methods for translation into daily use is currently under investigation.
Forty non-small-cell lung cancer paraffin-embedded samples with previous ALK (n = 33) and ROS1 (n = 7) FISH results were examined with the Oncomine Focus Assay and tested for ALK and ROS1 immunoreactivity. Clinical implications of concurrent molecular alterations and concordance between methods were evaluated.
NGS was successful in 32 (80%) cases: 25 ALK and 7 ROS1. Few concomitant alterations were detected: 1 ALK rearranged case had an ALK p.L1196M-resistant mutation, 4 had CDK4, MYC, and/or ALK amplifications, and 1 ROS1 rearranged case showed a FGFR4 amplification. Comparison between techniques revealed 5 (16%) discordant cases that had lower progression-free survival than concordant cases: 7.6 (95% confidence interval, 2.2-13) versus 19.4 (95% confidence interval, 10.1-28.6). Remarkably, 4 of these cases had isolated 3' signal FISH pattern (P = .026).
Our data support that the identification of 3' isolated signal FISH pattern in ALK and ROS1 cases might suggest a false-positive result. NGS seems a reliable technique to assess ALK and ROS1 rearrangements, offering the advantage over immunohistochemistry of detecting other molecular alterations with potential therapeutic implications.
检测非小细胞肺癌中的 ALK 和 ROS1 基因重排对于指导患者治疗至关重要。虽然荧光原位杂交（FISH）和免疫组织化学已被确立为金标准方法，但下一代测序（NGS）平台也被认为至少同样成功。目前正在研究将这些方法进行比较，以便将其转化为日常使用。
对 40 例先前有 ALK（n=33）和 ROS1（n=7）FISH 结果的非小细胞肺癌石蜡包埋样本进行了 Oncomine Focus 检测，并进行了 ALK 和 ROS1 免疫反应性检测。评估了同时存在的分子改变的临床意义以及方法之间的一致性。
NGS 在 32 例（80%）病例中成功：25 例 ALK 和 7 例 ROS1。检测到很少的伴随改变：1 例 ALK 重排病例存在 ALK p.L1196M 耐药突变，4 例存在 CDK4、MYC 和/或 ALK 扩增，1 例 ROS1 重排病例显示 FGFR4 扩增。技术比较显示 5 例（16%）不一致病例的无进展生存期比一致病例短：7.6（95%置信区间，2.2-13）与 19.4（95%置信区间，10.1-28.6）。值得注意的是，这些病例中有 4 例存在孤立的 3'信号 FISH 模式（P=0.026）。
我们的数据支持在 ALK 和 ROS1 病例中识别出孤立的 3'信号 FISH 模式可能提示假阳性结果。NGS 似乎是一种可靠的技术，可以评估 ALK 和 ROS1 重排，与免疫组织化学相比，它具有检测其他具有潜在治疗意义的分子改变的优势。
