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    7B7: a novel antibody directed against the Ku70/Ku80 heterodimer blocks invasion in pancreatic and lung cancer cells


    O'Sullivan, Dermot and Henry, Michael and Joyce, Helena and Walsh, Naomi and Mc Auley, Edel and Dowling, Paul and Swan, Niall and Moriarty, Michael and Barnham, Paul and Clynes, Martin and Larkin, Annemarie (2014) 7B7: a novel antibody directed against the Ku70/Ku80 heterodimer blocks invasion in pancreatic and lung cancer cells. Tumor Biology, 35 (7). pp. 6983-6997. ISSN 1010-4283

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    Abstract

    Development of more effective therapeutic strategies for cancers of high unmet need requires the continued discovery of disease-specific protein targets for therapeutic antibody targeting. In order to identify novel proteins associated with cancer cell invasion/metastasis, we present here an alternative to antibody targeting of cell surface proteins with an established role in invasion; our functional antibody screening approach involves the isolation and selection of MAbs that are primarily screened for their ability to inhibit tumour invasion. A clonal population of the Mia PaCa-2, a pancreatic ductal adenocarcinoma (PDAC) cell line, which displays a highly invasive phenotype, was used to generate MAbs with the objective of identifying membrane targets directly involved in cancer invasion. Selected MAb 7B7 can significantly reduce invasion in a dose-responsive manner in Mia PaCa-2 clone 3 and DLKP-M squamous lung carcinoma cells. Using immunoprecipitation and liquid chromatographytandem mass spectrometry (LC-MS-MS) analysis, the target antigen of anti-invasive antibody, 7B7, was determined to be the heterodimeric Ku antigen, Ku70/80, a core protein composed of the Ku70 and Ku80 subunits which is involved in non-homologous end-joining (NHEJ) DNA repair. RNA interference-mediated knockdown of Ku70 and Ku80 resulted in a marked decrease in the invasive capacity of Mia PaCa-2 clone 3 and DLKP-M cells, indicating that Ku70/Ku80 is functionally involved in pancreatic and lung cancer invasion. Immunohistochemical analysis demonstrated Ku70/Ku80 immunoreactivity in 37 PDAC tumours, indicating that this heterodimer is highly expressed in this aggressive cancer type. This study demonstrates that a functional MAb screening approach coupled with immunoprecipitation/proteomic analyses can be successfully applied to identify functional antiinvasive MAbs and potential novel targets for therapeutic antibody targeting.

    Item Type: Article
    Additional Information: The definitive published version of this article is available at DOI: 10.1007/s13277-014-1857-5
    Keywords: Monoclonal antibody; Cancermetastasis; Cancer invasion; Ku70/Ku80; Target discovery;
    Academic Unit: Faculty of Science and Engineering > Biology
    Item ID: 6927
    Identification Number: https://doi.org/10.1007/s13277-014-1857-5
    Depositing User: Paul Dowling
    Date Deposited: 26 Jan 2016 17:31
    Journal or Publication Title: Tumor Biology
    Publisher: Springer
    Refereed: Yes
    Funders: Enterprise Ireland
    URI:

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