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

Dermot O’Sullivan; Michael Henry; Helena Joyce; Naomi Walsh; Edel Mc Auley; Paul Dowling; Niall Swan; Michael Moriarty; Paul Barnham; Martin Clynes; Annemarie Larkin

doi:10.1007/s13277-014-1857-5

Tumor Biology

July 2014, Volume 35, Issue 7, pp 6983–6997

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

Authors
Authors and affiliations

Dermot O’Sullivan
Michael Henry
Helena Joyce
Naomi Walsh
Edel Mc Auley
Paul Dowling
Niall Swan
Michael Moriarty
Paul Barnham
Martin Clynes
Annemarie LarkinEmail author

Research Article

First Online:: 18 April 2014

Received:: 28 January 2014
Accepted:: 17 March 2014

1 Citations
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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 chromatography-tandem 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 anti-invasive MAbs and potential novel targets for therapeutic antibody targeting.

Keywords

Monoclonal antibody Cancer metastasis Cancer invasion Ku70/Ku80 Target discovery

Supplementary material

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Copyright information

Authors and Affiliations

Dermot O’Sullivan
- 1
- 2
Michael Henry
- 1
Helena Joyce
- 1
- 3
Naomi Walsh
- 1
Edel Mc Auley
- 1
Paul Dowling
- 1
Niall Swan
- 4
Michael Moriarty
- 1
Paul Barnham
- 5
Martin Clynes
- 1
Annemarie Larkin
- 1
Email author

1.National Institute for Cellular Biotechnology (NICB)Dublin City UniversityDublin 9Ireland
2.Biomedical Diagnostics InstituteDublin City UniversityDublin 9Ireland
3.Molecular Therapeutics Cancer Ireland (MTCI), National Institute for Cellular Biotechnology (NICB)Dublin City UniversityDublin 9Ireland
4.National Surgical Centre for Pancreatic CancerSt Vincent’s University HospitalDublinIreland
5.Dublin City UniversityDublin 9Ireland

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Cite this article as:: O’Sullivan, D., Henry, M., Joyce, H. et al. Tumor Biol. (2014) 35: 6983. doi:10.1007/s13277-014-1857-5

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

Abstract

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