Intravital imaging technology guides FAK-mediated priming in pancreatic cancer precision medicine according to Merlin status
Kendelle J. Murphy, Daniel A. Reed, Claire Vennin, James R. W. Conway, Max Nobis, Julia Yin, Cecilia R. Chambers, Brooke Pereira, Victoria Lee, Elysse C. Filipe, Michael Trpceski, Shona Ritchie, Morghan C. Lucas, Sean Warren, Joanna N. Skhinas, Astrid Magenau, Xanthe L. Metcalf, Janett Stoehr, Gretel Major, Ashleigh Parkin, Romain Bidanel, Ruth J. Lyons, Anaiis Zaratzian, Michael Tayao, Andrew Da Silva, Lea Abdulkhalek, Australian Pancreatic Genome Initiative (APGI), Australian Pancreatic Cancer Matrix Atlas (APMA), Anthony J. Gill, Amber L. Johns, Andrew V. Biankin, Jaswinder S. Samra, Sean M. Grimmond, Angela Chou, Jacky G. Goetz, Michael S. Samuel, J. Guy Lyons, Andrew Burgess, C. Elizabeth Caldon, Lisa G. Horvath, Roger J. Daly, Nikolaj Gadegaard, Yingxiao Wang, Owen J. Sansom, Jennifer P. Morton, Thomas R. Cox, Marina Pajic, David Herrmann, Paul Timpson
Abstract
Pancreatic ductal adenocarcinoma (PDAC) is a highly metastatic, chemoresistant malignancy and is characterized by a dense, desmoplastic stroma that modulates PDAC progression. Here, we visualized transient manipulation of focal adhesion kinase (FAK), which integrates bidirectional cell-environment signaling, using intravital fluorescence lifetime imaging microscopy of the FAK-based Förster resonance energy transfer biosensor in mouse and patient-derived PDAC models. Parallel real-time quantification of the FUCCI cell cycle reporter guided us to improve PDAC response to standard-of-care chemotherapy at primary and secondary sites. Critically, micropatterned pillar plates and stiffness-tunable matrices were used to pinpoint the contribution of environmental cues to chemosensitization, while fluid flow–induced shear stress assessment, patient-derived matrices, and personalized in vivo models allowed us to deconstruct how FAK inhibition can reduce PDAC spread. Last, stratification of PDAC patient samples via Merlin status revealed a patient subset with poor prognosis that are likely to respond to FAK priming before chemotherapy.