title: Imaging the impact of chemically inducible proteins on cellular dynamics in vivo.
abstract: The analysis of dynamic events in the tumor microenvironment during cancer progression is limited by the complexity of current in vivo imaging models. This is coupled with an inability to rapidly modulate and visualize protein activity in real time and to understand the consequence of these perturbations in vivo. We developed an intravital imaging approach that allows the rapid induction and subsequent depletion of target protein levels within human cancer xenografts while assessing the impact on cell behavior and morphology in real time. A conditionally stabilized fluorescent E-cadherin chimera was expressed in metastatic breast cancer cells, and the impact of E-cadherin induction and depletion was visualized using real-time confocal microscopy in a xenograft avian embryo model. We demonstrate the assessment of protein localization, cell morphology and migration in cells undergoing epithelial-mesenchymal and mesenchymal-epithelial transitions in breast tumors. This technique allows for precise control over protein activity in vivo while permitting the temporal analysis of dynamic biophysical parameters.
authors.affiliation: Translational Prostate Cancer Research Group, London Regional Cancer Program, London, Ontario, Canada.
authors.forename: Hon S
authors.initials: HS
authors.lastname: Leong
authors.forename: Michael M
authors.initials: MM
authors.lastname: Lizardo
authors.forename: Amber
authors.initials: A
authors.lastname: Ablack
authors.forename: Victor A
authors.initials: VA
authors.lastname: McPherson
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Wandless
authors.forename: Ann F
authors.initials: AF
authors.lastname: Chambers
authors.forename: John D
authors.initials: JD
authors.lastname: Lewis
mesh_terms: D000818:Animals; D001717:Birds / Q000196:embryology; D001943:Breast Neoplasms / Q000378:metabolism*; D015820:Cadherins / Q000378:metabolism*; D045744:Cell Line, Tumor; D003952:Diagnostic Imaging; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects; D005260:Female; D006801:Humans; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D009025:Morpholines / Q000493:pharmacokinetics / Q000494:pharmacology; D014183:Transplantation, Heterologous; D014746:Vimentin / Q000378:metabolism
chemical_list: D015820:Cadherins; D009025:Morpholines; C540198:Shield-1 compound; D014746:Vimentin