De Luca Caterina, Pepe Francesco, Iaccarino Antonino, Pisapia Pasquale, Righi Luisella, Listì Angela, Greco Lorenza, Gragnano Gianluca, Campione Severo, De Dominicis Gianfranco, Pagni Fabio, Sgariglia Roberta, Nacchio Mariantonia, Tufano Rossella, Conticelli Floriana, Vigliar Elena, Bellevicine Claudio, Cortinovis Diego Luigi, Novello Silvia, Molina-Vila Miguel Angel, Rosell Rafael, Troncone Giancarlo, Malapelle Umberto
Department of Public Health, University of Naples Federico II, 80131 Naples, Italy.
Department of Oncology, San Luigi University Hospital, University of Turin, 10043 Orbassano, Italy.
Cancers (Basel). 2021 Jan 4;13(1):139. doi: 10.3390/cancers13010139.
Gene fusions represent novel predictive biomarkers for advanced non-small cell lung cancer (NSCLC). In this study, we validated a narrow NGS gene panel able to cover therapeutically-relevant gene fusions and splicing events in advanced-stage NSCLC patients. To this aim, we first assessed minimal complementary DNA (cDNA) input and the limit of detection (LoD) in different cell lines. Then, to evaluate the feasibility of applying our panel to routine clinical samples, we retrospectively selected archived lung adenocarcinoma histological and cytological (cell blocks) samples. Overall, our SiRe RNA fusion panel was able to detect all fusions and a splicing event harbored in a RNA pool diluted up to 2 ng/µL. It also successfully analyzed 46 (95.8%) out of 48 samples. Among these, 43 (93.5%) out of 46 samples reproduced the same results as those obtained with conventional techniques. Intriguingly, the three discordant results were confirmed by a CE-IVD automated real-time polymerase chain reaction (RT-PCR) analysis (Easy PGX platform, Diatech Pharmacogenetics, Jesi, Italy). Based on these findings, we conclude that our new SiRe RNA fusion panel is a valid and robust tool for the detection of clinically relevant gene fusions and splicing events in advanced NSCLC.
基因融合是晚期非小细胞肺癌（NSCLC）新的预测性生物标志物。在本研究中，我们验证了一种窄谱NGS基因检测板，其能够覆盖晚期NSCLC患者中与治疗相关的基因融合和剪接事件。为此，我们首先评估了不同细胞系中最小互补DNA（cDNA）输入量和检测限（LoD）。然后，为了评估将我们的检测板应用于常规临床样本的可行性，我们回顾性选择了存档的肺腺癌组织学和细胞学（细胞块）样本。总体而言，我们的SiRe RNA融合检测板能够检测稀释至2 ng/µL的RNA池中所含的所有融合和一个剪接事件。它还成功分析了48个样本中的46个（95.8%）。其中，46个样本中的43个（93.5%）得到的结果与传统技术相同。有趣的是，这三个不一致的结果通过CE-IVD自动化实时聚合酶链反应（RT-PCR）分析（Easy PGX平台，Diatech Pharmacogenetics，意大利杰西）得到了证实。基于这些发现，我们得出结论，我们新的SiRe RNA融合检测板是检测晚期NSCLC中临床相关基因融合和剪接事件的有效且可靠的工具。
