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Updated: Aug 9, 2025
Characterizing Exon Skipping Efficiency in DMD Patient Samples in Clinical Trials of Antisense Oligonucleotides
Published on: May 7, 2020
An Italian Multicenter Perspective Harmonization Trial for the Assessment of MET Exon 14 Skipping Mutations in
Paolo Bironzo1, Francesco Pepe2, Gianluca Russo2
1Department of Oncology, S. Luigi Gonzaga Hospital, University of Turin, 10043 Orbassano, Italy.
Abstract:
Lung cancer remains the leading cause of cancer deaths worldwide. International societies have promoted the molecular analysis of MET proto-oncogene, receptor tyrosine kinase (MET) exon 14 skipping for the clinical stratification of non-small cell lung cancer (NSCLC) patients. Different technical approaches are available to detect MET exon 14 skipping in routine practice. Here, the technical performance and reproducibility of testing strategies for MET exon 14 skipping carried out in various centers were evaluated. In this retrospective study, each institution received a set (n = 10) of a customized artificial formalin-fixed paraffin-embedded (FFPE) cell line (Custom METex14 skipping FFPE block) that harbored the MET exon 14 skipping mutation (Seracare Life Sciences, Milford, MA, USA), which was previously validated by the Predictive Molecular Pathology Laboratory at the University of Naples Federico II. Each participating institution managed the reference slides according to their internal routine workflow. MET exon 14 skipping was successfully detected by all participating institutions. Molecular analysis highlighted a median Cq cut off of 29.3 (ranging from 27.1 to 30.7) and 2514 (ranging from 160 to 7526) read counts for real-time polymerase chain reaction (RT-PCR) and NGS-based analyses, respectively. Artificial reference slides were a valid tool to harmonize technical workflows in the evaluation of MET exon 14 skipping molecular alterations in routine practice.
Insights
Molecular testing for MET exon 14 skipping in non-small cell lung cancer (NSCLC) is crucial. Artificial reference slides effectively harmonized testing workflows across institutions, ensuring accurate detection of this important biomarker.
Area of Science:
- Oncology
- Molecular Pathology
- Genetics
Background:
- Lung cancer is a leading global cause of cancer mortality.
- Molecular analysis of MET exon 14 skipping mutations is vital for stratifying non-small cell lung cancer (NSCLC) patients.
- Standardized methods are needed to reliably detect MET exon 14 skipping in clinical practice.
Purpose of the Study:
- To evaluate the technical performance and reproducibility of different testing strategies for MET exon 14 skipping.
- To assess the utility of artificial formalin-fixed paraffin-embedded (FFPE) cell lines for harmonizing molecular testing workflows.
- To validate the detection of MET exon 14 skipping mutations across various laboratories.
Main Methods:
- A retrospective study involving multiple institutions.
- Distribution of customized artificial FFPE cell blocks harboring MET exon 14 skipping mutations.
- Analysis of MET exon 14 skipping using routine laboratory workflows, including real-time polymerase chain reaction (RT-PCR) and next-generation sequencing (NGS).
Main Results:
- All participating institutions successfully detected the MET exon 14 skipping mutation.
- Median Cq cutoff for RT-PCR was 29.3, and median read counts for NGS were 2514.
- Artificial reference slides proved effective in standardizing technical workflows for MET exon 14 skipping evaluation.
Conclusions:
- Artificial reference slides are a valuable tool for harmonizing molecular testing of MET exon 14 skipping in routine practice.
- Standardized detection of MET exon 14 skipping mutations across different centers is achievable.
- This approach supports consistent clinical stratification of NSCLC patients based on MET alterations.
