Gregory Weitsman
Detecting intratumoral heterogeneity of EGFR activity by liposome-based in vivo transfection of a fluorescent biosensor
Weitsman, Gregory; Mitchell, Nicholas J.; Evans, Rachel; Cheung, Anthony; Kalber, Tammy L.; Bofinger, Robin; Fruhwirth, Gilbert O.; Keppler, Melanie; Wright, Zoe V. F.; Barber, Paul R.; Gordon, Peter; Koning, Tamra de; Wulaningsih, Wahyu; Sander, Kerstin; Vojnovic, Borivoj; Hailes, Helen C.; Tabor, Alethea B.; Ng, Tony
Authors
Nicholas J. Mitchell
Rachel Evans
Anthony Cheung
Tammy L. Kalber
Robin Bofinger
Gilbert O. Fruhwirth
Melanie Keppler
Zoe V. F. Wright
Paul R. Barber
Peter Gordon
Tamra de Koning
Wahyu Wulaningsih
Kerstin Sander
Borivoj Vojnovic
Helen C. Hailes
Alethea B. Tabor
Tony Ng
Abstract
Despite decades of research in the epidermal growth factor receptor (EGFR) signalling field, and many targeted anti-cancer drugs that have been tested clinically, the success rate for these agents in the clinic is low, particularly in terms of the improvement of overall survival. Intratumoral heterogeneity is proposed as a major mechanism underlying treatment failure of these molecule-targeted agents. Here we highlight the application of fluorescence lifetime microscopy (FLIM)-based biosensing to demonstrate intratumoral heterogeneity of EGFR activity. For sensing EGFR activity in cells, we used a genetically encoded CrkII-based biosensor which undergoes conformational changes upon tyrosine-221 phosphorylation by EGFR. We transfected this biosensor into EGFR-positive tumour cells using targeted lipopolyplexes bearing EGFR-binding peptides at their surfaces. In a murine model of basal-like breast cancer, we demonstrated a significant degree of intratumoral heterogeneity in EGFR activity, as well as the pharmacodynamic effect of a radionuclide-labeled EGFR inhibitor in situ. Furthermore, a significant correlation between high EGFR activity in tumour cells and macrophage-tumour cell proximity was found to in part account for the intratumoral heterogeneity in EGFR activity observed. The same effect of macrophage infiltrate on EGFR activation was also seen in a colorectal cancer xenograft. In contrast, a non-small cell lung cancer xenograft expressing a constitutively active EGFR conformational mutant exhibited macrophage proximity-independent EGFR activity. Our study validates the use of this methodology to monitor therapeutic response in terms of EGFR activity. In addition, we found iNOS gene induction in macrophages that are cultured in tumour cell-conditioned media as well as an iNOS activity-dependent increase in EGFR activity in tumour cells. These findings point towards an immune microenvironment-mediated regulation that gives rise to the observed intratumoral heterogeneity of EGFR signalling activity in tumour cells in vivo.
Citation
Weitsman, G., Mitchell, N. J., Evans, R., Cheung, A., Kalber, T. L., Bofinger, R., Fruhwirth, G. O., Keppler, M., Wright, Z. V. F., Barber, P. R., Gordon, P., Koning, T. D., Wulaningsih, W., Sander, K., Vojnovic, B., Hailes, H. C., Tabor, A. B., & Ng, T. (in press). Detecting intratumoral heterogeneity of EGFR activity by liposome-based in vivo transfection of a fluorescent biosensor. Oncogene, https://doi.org/10.1038/onc.2016.522
Journal Article Type | Article |
---|---|
Acceptance Date | Dec 21, 2016 |
Online Publication Date | Feb 6, 2017 |
Deposit Date | Mar 1, 2017 |
Publicly Available Date | Mar 1, 2017 |
Journal | Oncogene |
Print ISSN | 0950-9232 |
Electronic ISSN | 1476-5594 |
Publisher | Nature Publishing Group |
Peer Reviewed | Peer Reviewed |
DOI | https://doi.org/10.1038/onc.2016.522 |
Public URL | https://nottingham-repository.worktribe.com/output/847566 |
Publisher URL | http://www.nature.com/onc/journal/vaop/ncurrent/full/onc2016522a.html |
Contract Date | Mar 1, 2017 |
Files
onc2016522a.pdf
(2.3 Mb)
PDF
Copyright Statement
Copyright information regarding this work can be found at the following address: http://creativecommons.org/licenses/by/4.0
You might also like
Downloadable Citations
About Repository@Nottingham
Administrator e-mail: discovery-access-systems@nottingham.ac.uk
This application uses the following open-source libraries:
SheetJS Community Edition
Apache License Version 2.0 (http://www.apache.org/licenses/)
PDF.js
Apache License Version 2.0 (http://www.apache.org/licenses/)
Font Awesome
SIL OFL 1.1 (http://scripts.sil.org/OFL)
MIT License (http://opensource.org/licenses/mit-license.html)
CC BY 3.0 ( http://creativecommons.org/licenses/by/3.0/)
Powered by Worktribe © 2025
Advanced Search