Visualizing Ligand Binding to a GPCR In Vivo Using NanoBRET

Diana C Alcobia; Alexandra I Ziegler; Alexander Kondrashov; Eleonora Comeo; Sarah Mistry; Barrie Kellam; Aeson Chang; Jeanette Woolard; Stephen J Hill; Erica K Sloan

doi:10.1016/j.isci.2018.08.006

Visualizing Ligand Binding to a GPCR In Vivo Using NanoBRET

iScience. 2018 Aug 31:6:280-288. doi: 10.1016/j.isci.2018.08.006. Epub 2018 Aug 11.

Authors

Affiliations

¹ Division of Physiology, Pharmacology and Neuroscience, School of Life Sciences, University of Nottingham, Nottingham NG7 2UH, UK; Centre of Membrane Proteins and Receptors, University of Birmingham and University of Nottingham, The Midlands, UK; Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, VIC 3052, Australia.
² Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, VIC 3052, Australia.
³ Wolfson Centre for Stem Cells, Tissue Engineering & Modelling (STEM), Centre for Biomolecular Sciences, University of Nottingham, Nottingham NG7 2RD, UK.
⁴ Centre of Membrane Proteins and Receptors, University of Birmingham and University of Nottingham, The Midlands, UK; School of Pharmacy, Division of Biomolecular Science and Medicinal Chemistry, Centre for Biomolecular Sciences, University of Nottingham, Nottingham NG7 2RD, UK.
⁵ Division of Physiology, Pharmacology and Neuroscience, School of Life Sciences, University of Nottingham, Nottingham NG7 2UH, UK; Centre of Membrane Proteins and Receptors, University of Birmingham and University of Nottingham, The Midlands, UK. Electronic address: jeanette.woolard@nottingham.ac.uk.
⁶ Division of Physiology, Pharmacology and Neuroscience, School of Life Sciences, University of Nottingham, Nottingham NG7 2UH, UK; Centre of Membrane Proteins and Receptors, University of Birmingham and University of Nottingham, The Midlands, UK. Electronic address: steve.hill@nottingham.ac.uk.
⁷ Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, VIC 3052, Australia; Cousins Center for Neuroimmunology, Semel Institute for Neuroscience and Human Behavior, Jonsson Comprehensive Cancer Center, and UCLA AIDS Institute, University of California Los Angeles, Los Angeles, CA 90095, USA; Division of Surgical Oncology, Peter MacCallum Cancer Centre, Victorian Comprehensive Cancer Centre, 305 Grattan Street, Melbourne, VIC 3000, Australia. Electronic address: erica.sloan@monash.edu.

Abstract

The therapeutic action of a drug depends on its ability to engage with its molecular target in vivo. However, current drug discovery strategies quantify drug levels within organs rather than determining the binding of drugs directly to their specific molecular targets in vivo. This is a particular problem for assessing the therapeutic potential of drugs that target malignant tumors where access and binding may be impaired by disrupted vasculature and local hypoxia. Here we have used triple-negative human breast cancer cells expressing β₂-adrenoceptors tagged with the bioluminescence protein NanoLuc to provide a bioluminescence resonance energy transfer approach to directly quantify ligand binding to a G protein-coupled receptor in vivo using a mouse model of breast cancer.

Keywords: Biological Sciences Tools; Cancer; Molecular Interaction; Optical Imaging.

Abstract

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