Intravital Imaging to Monitor Therapeutic Response in Moving Hypoxic Regions Resistant to PI3K Pathway Targeting in Pancreatic Cancer

James R W Conway; Sean C Warren; David Herrmann; Kendelle J Murphy; Aurélie S Cazet; Claire Vennin; Robert F Shearer; Monica J Killen; Astrid Magenau; Pauline Mélénec; Mark Pinese; Max Nobis; Anaiis Zaratzian; Alice Boulghourjian; Andrew M Da Silva; Gonzalo Del Monte-Nieto; Arne S A Adam; Richard P Harvey; Jody J Haigh; Yingxiao Wang; David R Croucher; Owen J Sansom; Marina Pajic; C Elizabeth Caldon; Jennifer P Morton; Paul Timpson

doi:10.1016/j.celrep.2018.05.038

Intravital Imaging to Monitor Therapeutic Response in Moving Hypoxic Regions Resistant to PI3K Pathway Targeting in Pancreatic Cancer

Cell Rep. 2018 Jun 12;23(11):3312-3326. doi: 10.1016/j.celrep.2018.05.038.

Authors

James R W Conway¹, Sean C Warren¹, David Herrmann², Kendelle J Murphy¹, Aurélie S Cazet¹, Claire Vennin¹, Robert F Shearer¹, Monica J Killen¹, Astrid Magenau², Pauline Mélénec¹, Mark Pinese², Max Nobis¹, Anaiis Zaratzian¹, Alice Boulghourjian¹, Andrew M Da Silva¹, Gonzalo Del Monte-Nieto³, Arne S A Adam⁴, Richard P Harvey⁵, Jody J Haigh⁶, Yingxiao Wang⁷, David R Croucher⁸, Owen J Sansom⁹, Marina Pajic², C Elizabeth Caldon², Jennifer P Morton¹⁰, Paul Timpson¹¹

Affiliations

¹ Garvan Institute of Medical Research and The Kinghorn Cancer Centre, Cancer Division, Sydney, NSW 2010, Australia.
² Garvan Institute of Medical Research and The Kinghorn Cancer Centre, Cancer Division, Sydney, NSW 2010, Australia; St Vincent's Clinical School, Faculty of Medicine, University of NSW, Sydney, NSW 2010, Australia.
³ St Vincent's Clinical School, Faculty of Medicine, University of NSW, Sydney, NSW 2010, Australia; Developmental and Stem Cell Biology Division, Victor Chang Cardiac Research Institute, Sydney, NSW 2010, Australia.
⁴ Developmental and Stem Cell Biology Division, Victor Chang Cardiac Research Institute, Sydney, NSW 2010, Australia.
⁵ St Vincent's Clinical School, Faculty of Medicine, University of NSW, Sydney, NSW 2010, Australia; Developmental and Stem Cell Biology Division, Victor Chang Cardiac Research Institute, Sydney, NSW 2010, Australia; School of Biotechnology and Biomolecular Science, University of New South Wales, Sydney, NSW 2033, Australia.
⁶ Australian Centre for Blood Diseases, Monash University, Melbourne, VIC 3004, Australia.
⁷ Department of Bioengineering, University of Illinois, Urbana-Champaign, Urbana, IL 61801, USA.
⁸ Garvan Institute of Medical Research and The Kinghorn Cancer Centre, Cancer Division, Sydney, NSW 2010, Australia; St Vincent's Clinical School, Faculty of Medicine, University of NSW, Sydney, NSW 2010, Australia; School of Medicine and Medical Science, University College Dublin, Belfield, Dublin 4, Ireland.
⁹ Cancer Research UK Beatson Institute, Switchback Road, Bearsden, Glasgow G61 1BD, UK; Institute of Cancer Sciences, University of Glasgow, Glasgow G61 1QH, UK.
¹⁰ Cancer Research UK Beatson Institute, Switchback Road, Bearsden, Glasgow G61 1BD, UK; Institute of Cancer Sciences, University of Glasgow, Glasgow G61 1QH, UK. Electronic address: j.morton@beatson.gla.ac.uk.
¹¹ Garvan Institute of Medical Research and The Kinghorn Cancer Centre, Cancer Division, Sydney, NSW 2010, Australia; St Vincent's Clinical School, Faculty of Medicine, University of NSW, Sydney, NSW 2010, Australia. Electronic address: p.timpson@garvan.org.au.

Abstract

Application of advanced intravital imaging facilitates dynamic monitoring of pathway activity upon therapeutic inhibition. Here, we assess resistance to therapeutic inhibition of the PI3K pathway within the hypoxic microenvironment of pancreatic ductal adenocarcinoma (PDAC) and identify a phenomenon whereby pronounced hypoxia-induced resistance is observed for three clinically relevant inhibitors. To address this clinical problem, we have mapped tumor hypoxia by both immunofluorescence and phosphorescence lifetime imaging of oxygen-sensitive nanoparticles and demonstrate that these hypoxic regions move transiently around the tumor. To overlay this microenvironmental information with drug response, we applied a FRET biosensor for Akt activity, which is a key effector of the PI3K pathway. Performing dual intravital imaging of drug response in different tumor compartments, we demonstrate an improved drug response to a combination therapy using the dual mTORC1/2 inhibitor AZD2014 with the hypoxia-activated pro-drug TH-302.

Keywords: AZD2014; Akt; FRET; PI3K pathway; PLIM; hypoxia; intravital imaging; nanoparticles; pancreatic cancer; pro-drug.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Benzamides
Cell Line, Tumor
Drug Resistance, Neoplasm* / drug effects
Drug Therapy, Combination
Female
Fluorescence Resonance Energy Transfer
Humans
Hypoxia
Intravital Microscopy / methods
Mice
Mice, Inbred BALB C
Morpholines / pharmacology
Morpholines / therapeutic use
Nanoparticles / chemistry
Nitroimidazoles / pharmacology
Nitroimidazoles / therapeutic use
Pancreatic Neoplasms / drug therapy
Pancreatic Neoplasms / metabolism
Pancreatic Neoplasms / pathology
Phosphatidylinositol 3-Kinases / metabolism*
Phosphoramide Mustards / pharmacology
Phosphoramide Mustards / therapeutic use
Proto-Oncogene Proteins c-akt / metabolism
Pyrimidines
Signal Transduction* / drug effects
Transplantation, Heterologous
Tumor Microenvironment

Substances

Benzamides
Morpholines
Nitroimidazoles
Phosphoramide Mustards
Pyrimidines
TH 302
vistusertib
Proto-Oncogene Proteins c-akt

Grants and funding

21139/CRUK_/Cancer Research UK/United Kingdom