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
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: |
|
| Use Licence: |
This item is available under a Creative Commons Attribution Non Commercial Share Alike Licence (CC BY-NC-SA). Details of this licence are available
here |
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