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Donald Green



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    P3.09 - Pathology (Not CME Accredited Session) (ID 975)

    • Event: WCLC 2018
    • Type: Poster Viewing in the Exhibit Hall
    • Track:
    • Presentations: 1
    • Moderators:
    • Coordinates: 9/26/2018, 12:00 - 13:30, Exhibit Hall
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      P3.09-18 - Identification of MET exon 14 Skipping Mutations by FusionPlex<sup>TM</sup> Solid Tumor Panel (ID 14204)

      12:00 - 13:30  |  Author(s): Donald Green

      • Abstract

      Background

      MET exon 14 skipping (METex14), or splice site, mutations result in the deletion of the juxtamembrane domain of MET, leading to increased MET receptor pathway signaling. Non-small cell lung cancer (NSCLC) patients with METex14 mutations are eligible for therapy with MET inhibitors like crizotinib. METex14 mutations are diverse and complex and can be challenging to detect by targeted DNA NGS assays alone. The Archer FusionPlexTM Solid Tumor panel use anchored multiplex PCR chemistry to detect fusions and other mutations including oncogenic isoforms in 53 genes associated with solid tumors.

      a9ded1e5ce5d75814730bb4caaf49419 Method

      In July 2016, we implemented the FusionPlex panel in our clinical laboratory for routine testing of NSCLC. RNA is extracted from FFPE tissue using the AllPrep DNA/RNA FFPE Kit and RNA quantity is assessed using the Qubit HS RNA Assay, and RNA quality is assessed using the PreSeq RNA QC Kit. cDNA synthesis and library preparation are performed according to the manufacturer’s recommendations. Libraries are quantified using the KAPA Library Quantification Kit, diluted to 4 nM, pooled and sequenced on the Illumina MiSeq System. Data analysis is performed using the Archer Analysis platform (v4.0.11). MET exon 14 isoforms were confirmed by an orthogonal method (DNA NGS assay and/or melt curve analysis).

      4c3880bb027f159e801041b1021e88e8 Result

      Since July 2016, samples from 275 NSCLC patients were successfully sequenced. Six (2.2%) MET oncogenic isoforms, consistent with MET exon 14 skipping mutations, were identified and subsequently confirmed. All six patients were female with an age range of 61-82 years. Two presented with stage I disease, two stage II, one stage III and one stage IV. To date, none of the patients have been treated with MET inhibitor therapy.

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      8eea62084ca7e541d918e823422bd82e Conclusion

      The FusionPlexTM Solid Tumor NGS panel can be routinely performed in the clinical laboratory to optimize detection of MET exon 14 mutations as well as other clinically actionable gene fusions.

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