Related Experiment Video
Updated: Jun 11, 2025
Author Spotlight: Advancements in Molecular Biomarker Testing for Non-Squamous Non-Small Cell Lung Cancer
Published on: September 8, 2023
Clinical utility of ctDNA by amplicon based next generation sequencing in first line non small cell lung cancer
Valerio Gristina1, Tancredi Didier Bazan Russo1, Nadia Barraco1
1Department of Precision Medicine in Medical, Surgical and Critical Care (Me.Pre.C.C.), University of Palermo, Palermo, Italy.
Abstract:
The assessment of ctDNA has emerged as a minimally invasive avenue for molecular diagnosis and real-time tracking of tumor progression in NSCLC. However, the evaluation of ctDNA by amplicon-based NGS has been not endorsed by all the healthcare systems and remains to be fully integrated into clinical routine practice. To compare tissue single-gene with plasma multiplexed testing, we retrospectively evaluated 120 plasma samples from 12 consecutive patients with advanced non-squamous NSCLC who were part of a prospective study enrolling treatment-naïve patients and in which tissue samples were evaluated using a single-gene testing approach. While the plasma ctDNA detection of EGFR and BRAF mutations had an acceptable level of concordance with the archival tissue (85%), discordance was seen in all the patients in whom ALK alterations were only detected in tissue samples. Among six responders and six non-responders, early ctDNA mutant allelic frequency (MAF) reduction seemed to predict radiologic responses and longer survival, whereas increasing MAF values with the emergence of co-mutations like BRAFV600E, KRASG12V or TP53M237I seemed to be an early indicator of molecular and radiologic progression. This report using an amplicon-based NGS assay on ctDNA underscores the real-life need for plasma and tissue genotyping as complementary tools in the diagnostic and therapeutic decision-making process.
Insights
Circulating tumor DNA (ctDNA) testing shows promise for non-small cell lung cancer (NSCLC) management. Plasma and tissue genotyping are complementary tools for diagnosis and treatment decisions.
Area of Science:
- Oncology
- Molecular Diagnostics
- Genetics
Background:
- Circulating tumor DNA (ctDNA) offers a minimally invasive method for molecular diagnosis and monitoring of non-small cell lung cancer (NSCLC).
- Amplicon-based next-generation sequencing (NGS) for ctDNA evaluation is not universally adopted in healthcare systems and requires further clinical integration.
Purpose of the Study:
- To compare the concordance of plasma ctDNA multiplexed testing with tissue single-gene testing in advanced non-squamous NSCLC patients.
- To evaluate the utility of ctDNA mutant allelic frequency (MAF) dynamics in predicting treatment response and survival.
Main Methods:
- Retrospective analysis of 120 plasma samples from 12 treatment-naïve advanced non-squamous NSCLC patients.
- Comparison of plasma ctDNA results with archival tissue samples tested via single-gene assays.
- Assessment of ctDNA MAF changes in relation to treatment response and survival outcomes.
Main Results:
- Plasma ctDNA detection of EGFR and BRAF mutations showed 85% concordance with tissue samples.
- Complete discordance was observed for ALK alterations, exclusively detected in tissue samples.
- Early reduction in ctDNA MAF correlated with radiologic response and improved survival.
- Increasing ctDNA MAF and emergence of co-mutations indicated molecular and radiologic progression.
Conclusions:
- Plasma and tissue genotyping serve as complementary tools in the diagnostic and therapeutic decision-making process for NSCLC.
- Amplicon-based NGS assays on ctDNA highlight the clinical need for integrated plasma and tissue molecular profiling.
