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  P2.06 - Mesothelioma (Not CME Accredited Session) (ID 955)

  • Event: WCLC 2018
  • Type: Poster Viewing in the Exhibit Hall
  • Track:
  • Presentations: 1
  • Moderators:
  • Coordinates: 9/25/2018, 16:45 - 18:00, Exhibit Hall

  • 27015

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    P2.06-07 - Genomic Deletion of BAP1 and CDKN2A are Better MM Diagnostic Biomarkers.  (ID 13623)

    16:45 - 18:00  |  Author(s): Kenneth Lee
    • Abstract
    • Slides

    Background
    

    Malignant mesothelioma (MM) is deadly cancer caused by asbestos exposure and has limited treatment options. More reliable MM biomarkers and more accurate detection methods are urgent requirements for early identification of this cancer. MM cell cultures are essential in vitro models for potential treatment options analysis. In this study, we established primary MM cell lines and characterised them with current biomarkers as well as utilising the latest digital droplet PCR instrument for genomic deletion analysis. We aim to discover better diagnostic biomarkers for detection of MM.

    a9ded1e5ce5d75814730bb4caaf49419 Method
    

    Pathologically confirmed MM samples from patients were used to establish MM cell lines. MM cells were grown over 20 passages to ensure homogenous MM cell populations. Original MM tissue samples and established MM cells were stained with H&E and verified with a diagnostic biomarker panel (CK-8/18, Calretinin, CK5/6, CD141, WT-1, D2-40, EMA, CEA, Tag-72, BG8, CD15, TTF-1 and BAP1). MM cells were grown in 2D and 3D conformations for biomarker staining and compared to stained tissue biomarkers. Scores and comments were provided by pathologists experienced in MM diagnosis. Laser Capture micro-dissected (LCM) MM samples and established cells were used for DNA isolation. Genomic deletions were analysed by digital droplet PCR for copy number variations of BAP1 and CDKN2A.

    4c3880bb027f159e801041b1021e88e8 Result
    

    15 MM cell lines established from original tissue were grown over 20 passages which produced homogeneous cell populations under normal cell culture conditions. Cells grown in 3D with H&E staining exhibited better tumour architecture, cell-to-cell contacts and tumour cell-like morphology when compared to cells grown in standard 2D culture. The percentage of MM biomarker levels showed variation between original tissue and cells established in culture. Genomic deletions of BAP1 and CDKN2A, however, showed consistent levels between the two.

    8eea62084ca7e541d918e823422bd82e Conclusion
    

    Conventional immunohistochemistry MM biomarkers did not show sustained correlation between tissues of origin to established cell culture, whereas genomic deletion analysis did. Genomic deletion analysis has the potential to be used as more accurate biomarkers for MM diagnosis.

    6f8b794f3246b0c1e1780bb4d4d5dc53

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