Author of

• 35813

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  P62 - Tumor Biology and Systems Biology - Basic and Translational Science - Metabolomics (ID 200)

  • Event: WCLC 2020
  • Type: Posters
  • Track: Tumor Biology and Systems Biology - Basic and Translational Science
  • Presentations: 1
  • Moderators:
  • Coordinates: 1/28/2021, 00:00 - 00:00, ePoster Hall

  • 35815

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    P62.02 - Identification of Essential Genes in Malignant Pleural Mesothelioma Using Genome-Wide CRISPR/Cas9-Based Negative Selection Screen (ID 2401)

    00:00 - 00:00  |  Presenting Author(s): Ece Cakiroglu
    • Abstract
    • Slides

    Introduction
    

    Malignant pleural mesothelioma (MPM) is a rare cancer with an increasing incidence and low survival rates. Existing treatment options are limited to chemotherapy and/or multimodal therapy with poor prognosis. Therefore, novel targeted therapies are needed. Although many gene mutations have been identified so far, the absence of a defined oncogene dependency limits the targeted treatment options.

    Methods
    

    In the present study, we employed genome-wide negative selection CRISPR-Cas9 screening to identify cancer cell essential genes and drugable targets in MPM. To this end, we engineered clonal sublines with permanent Cas9 nuclease expression in a normal mesothelial cell line (MeT-5A) and 3 MPM cell lines (H2052, H2452, and H28). For this aim, target cells were initially transduced with pLentiCas9-EGFP lentiviral vector and the highest eGFP expressing cells were FACS-sorted and further processed to generate single cell clones with consistent, uniform and high-level Cas9 expression. In addition, Cas9 protein expression and genome editing efficiencies were validated accordingly. Afterwards, we screened cells by utilizing Brunello genome-wide CRISPR knock-out library.

    Results
    

    Not surprisingly, sequencing results showed hits in common essential genes that take role in replication, translation, and transcription and in genes that have been previously shown to be mutated in MPM e.g. BAP1, WEE1, FGFR1. Our results demonstrate that MPM cell lines show sensitivity to depletion of RNF8 and XRCC3 DNA repair genes when compared to normal mesothelial cell line.

    Conclusion
    

    We anticipate that, further analysis and validations will reveal vulnerabilities and possible drugable targets in MPM.

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