Visualizing Ligand Binding to a GPCR In Vivo Using NanoBRET

• Published in: iScience 2018-08, Vol.6, p.280-288
• Main Authors: Alcobia, Diana C., Ziegler, Alexandra I., Kondrashov, Alexander, Comeo, Eleonora, Mistry, Sarah, Kellam, Barrie, Chang, Aeson, Woolard, Jeanette, Hill, Stephen J., Sloan, Erica K.
• Format: Article
• Language: English
• Subjects: Biological Sciences Tools; Cancer; Molecular Interaction; Optical Imaging
• ISSN: 2589-0042; 2589-0042
• DOI: 10.1016/j.isci.2018.08.006

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. [Display omitted] •NanoLuc luciferase was used to localize β2-adrenoceptors in vivo•Metastatic breast cancer cells retain expression of β2-adrenoceptors•The binding of fluorescent propranolol to β2-adrenoceptors was quantified in vivo•NanoBRET was used to monitor target engagement in a mouse model of breast cancer Biological Sciences Tools; Cancer; Molecular Interaction; Optical Imaging