Login (DCU Staff Only)
Login (DCU Staff Only)

DORAS | DCU Research Repository

Explore open access research and scholarly works from DCU

Advanced Search

Tyrosine kinase inhibitors as modulators of trastuzumab-mediated antibody dependent cell-mediated cytotoxicity in breast cancer cell lines

Collins, Denis orcid logoORCID: 0000-0003-1043-6169, Gately, Kathy orcid logoORCID: 0000-0002-9616-424X, Hughes, Clare, Edwards, Connla, Davies, Anthony, Madden, Stephen F. orcid logoORCID: 0000-0002-0489-4588, O'Byrne, Kenneth J., O'Donovan, Norma orcid logoORCID: 0000-0003-1113-3872 and Crown, John orcid logoORCID: 0000-0002-3125-7613 (2017) Tyrosine kinase inhibitors as modulators of trastuzumab-mediated antibody dependent cell-mediated cytotoxicity in breast cancer cell lines. Cellular Immunology, 17 (2). pp. 35-42. ISSN 0008-8749

Abstract
Background: Trastuzumab is an anti-HER2 monoclonal antibody (mAb) therapy capable of antibody-dependent cell-mediated cytotoxicity (ADCC) and used in the treatment of HER2+ breast cancer. Through interactions with FcƴR+ immune cell subsets, trastuzumab functions as a passive immunotherapy. The EGFR/HER2-targeting tyrosine kinase inhibitor (TKI) lapatinib and the next generation TKIs afatinib and neratinib, can alter HER2 levels, potentially modulating the ADCC response to trastuzumab. Using LDHrelease assays, we investigated the impact of antigen modulation, assay duration and peripheral blood mononuclear cell (PBMC) activity on trastuzumab-mediated ADCC in breast cancer models of maximal (SKBR3) and minimal (MCF-7) target antigen expression to determine if modulating the ADCC response to trastuzumab using TKIs may be a viable approach for enhancing tumor immune reactivity. Methods: HER2 levels were determined in lapatinib, afatinib and neratinib-treated SKBR3 and MCF-7 using high content analysis (HCA). Trastuzumab-mediated ADCC was assessed following treatment with TKIs utilising a colorimetric LDH release-based protocol at 4 and 12 hour timepoints. PBMC activity was assessed against non-MHCrestricted K562 cells. A flow cytometry-based method (CFSE/7-AAD) was also used to measure trastuzumab-mediated ADCC in medium-treated SKBR3 and MCF-7. Results: HER2 antigen levels were significantly altered by the three TKIs in both cell line models. The TKIs significantly reduced LDH levels directly in SKBR3 cells but not MCF-7. Lapatinib and neratinib augment trastuzumab-related ADCC in SKBR3 but the effect was not consistent with antigen expression levels and was dependent on volunteer PBMC activity (vs. K562). A 12 hour assay timepoint produced more consistent results. 3 Trastuzumab-mediated ADCC (PBMC:target cell ratio of 10:1) was measured at 7.6 ± 4.7% (T12) by LDH assay and 19 ± 3.2 % (T12) using the flow cytometry-based method in the antigen-low model MCF-7.Conclusions:In the presence of effector cells with high cytotoxic capacity, TKIs have the ability to augment the passive immunotherapeutic potential of trastuzumab in SKBR3, a model of HER2+ breast cancer. ADCC levels detected by LDH-release assays are extremely low in MCF-7; the flow cytometry-based CFSE/7-AAD method is more sensitive and consistent for the determination of ADCC in HER2-low models.
Metadata
Item Type:Article (Published)
Refereed:Yes
Uncontrolled Keywords:Monoclonal antibody therapy; ADCC; trastuzumab; HER2; tyrosine kinase inhibitors; LDH assay
Subjects:Medical Sciences > Cancer
DCU Faculties and Centres:DCU Faculties and Schools > Faculty of Science and Health > School of Biotechnology
Research Initiatives and Centres > National Institute for Cellular Biotechnology (NICB)
Publisher:Elsevier
Official URL:https://doi.org/10.1016/j.cellimm.2017.07.005
Copyright Information:© 2017 Elsevier
Funders:Irish Research Council Enterprise Partnership Scheme, Roche Products Ireland Ltd, Cancer Clinical Research Trust (CHY12210), Science Foundation Ireland funded Molecular Therapeutics for Cancer Ireland (08/SRC/B1410
ID Code:29591
Deposited On:26 Feb 2024 17:03 by Thomas Murtagh . Last Modified 26 Feb 2024 17:03
Documents

Full text available as:

[thumbnail of Collins DM et al 2017 TKI ADCC PMID 28735814 Author version Upload 2024.pdf]
Preview
PDF - Requires a PDF viewer such as GSview, Xpdf or Adobe Acrobat Reader
Creative Commons: Attribution-Noncommercial-No Derivative Works 4.0
1MB
Downloads

Downloads

Downloads per month over past year

Archive Staff Only: edit this record