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Updated: Nov 22, 2025
Author Spotlight: Advancements in Molecular Biomarker Testing for Non-Squamous Non-Small Cell Lung Cancer
Published on: September 8, 2023
RNA-Based Assay for Next-Generation Sequencing of Clinically Relevant Gene Fusions in Non-Small Cell Lung Cancer
Caterina De Luca1, Francesco Pepe1, Antonino Iaccarino1
1Department of Public Health, University of Naples Federico II, 80131 Naples, Italy.
Abstract:
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.
Insights
A new RNA fusion panel effectively detects therapeutically relevant gene fusions and splicing events in non-small cell lung cancer (NSCLC) patients. This validated NGS panel shows promise for routine clinical use in advanced NSCLC diagnostics.
Area of Science:
- Oncology
- Molecular Diagnostics
- Genomics
Background:
- Gene fusions are emerging as critical predictive biomarkers in advanced non-small cell lung cancer (NSCLC).
- Accurate detection of these biomarkers is essential for guiding targeted therapies in NSCLC patients.
Purpose of the Study:
- To validate a targeted next-generation sequencing (NGS) gene panel for detecting therapeutically relevant gene fusions and splicing events.
- To assess the panel's performance using minimal RNA input and its feasibility in routine clinical settings for advanced NSCLC.
Main Methods:
- Assessed minimal complementary DNA (cDNA) input and limit of detection (LoD) using cell lines.
- Retrospectively analyzed archived lung adenocarcinoma histological and cytological samples.
- Validated the SiRe RNA fusion panel against conventional techniques and RT-PCR for discordant results.
Main Results:
- The SiRe RNA fusion panel detected all target fusions and a splicing event in RNA diluted down to 2 ng/µL.
- The panel successfully analyzed 95.8% of retrospective clinical samples (46/48).
- High concordance (93.5%) was observed with conventional methods, with discordant results confirmed by RT-PCR.
Conclusions:
- The SiRe RNA fusion panel is a validated and robust tool for detecting clinically significant gene fusions and splicing events in advanced NSCLC.
- This NGS panel demonstrates potential for routine clinical application in NSCLC diagnostics.
- The findings support the use of this panel for identifying patients who may benefit from targeted therapies.
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