Type

Journal Article

Authors

Annemarie Larkin
Martin Clynes
Paul Barnham
Michael Moriarty
Niall Swan
Paul Dowling
Edel Mc Auley
Naomi Walsh
Helena Joyce
Michael Henry
and 1 others

Subjects

Microbiology

Topics
antigens nuclear mass spectrometry lc ms novel the administration dosage lung neoplasms cell line dna binding proteins ku autoantigen immunotherapy cancer invasion therapy pancreatic cancer humans immunology chemistry genetics antibodies monoclonal proteomics pathology protein multimerization pancreatic neoplasms lung cancer cells rna interference neoplasm invasiveness lung carcinoma cells cell line tumor

7B7: a novel antibody directed against the Ku70/Ku80 heterodimer blocks invasion in pancreatic and lung cancer cells. (2014)

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.
Collections Ireland -> Maynooth University -> Academic Unit = Faculty of Science and Engineering: Biology
Ireland -> Maynooth University -> Type = Article
Ireland -> Dublin City University -> PubMed
Ireland -> Maynooth University -> Academic Unit = Faculty of Science and Engineering
Ireland -> Maynooth University -> Status = Published
Ireland -> Maynooth University -> Open Access DRIVERset

Full list of authors on original publication

Annemarie Larkin, Martin Clynes, Paul Barnham, Michael Moriarty, Niall Swan, Paul Dowling, Edel Mc Auley, Naomi Walsh, Helena Joyce, Michael Henry and 1 others

Experts in our system

1
Annemarie Larkin
Dublin City University
Total Publications: 25
 
2
Martin Clynes
Dublin City University
Total Publications: 209
 
3
Michael Moriarty
Dublin City University
Total Publications: 8
 
4
N Swan
IT Tallaght
Total Publications: 19
 
5
Paul Dowling
Dublin City University
Total Publications: 79
 
6
Naomi Walsh
Dublin City University
Total Publications: 18
 
7
Helena Joyce
Dublin City University
Total Publications: 10
 
8
Michael Henry
Dublin City University
Total Publications: 78