Development of High-Specificity Fluorescent Probes to Enable Cannabinoid Type 2 Receptor Studies in Living Cells
Year: 2020
Type: article
Abstract: Pharmacological modulation of cannabinoid type 2 receptor (CB2R) holds promise for the treatment of numerous conditions, including inflammatory diseases, autoimmune disorders, pain, and cancer. Despite the significance of this receptor, researchers lack reliable tools to address questions concerning the expression and complex mechanism of CB2R signaling, especially in cell-type and tissue-dependent contexts. Herein, we report for the first time a versatile ligand platform for the modular design of a collection of highly specific CB2R fluorescent probes, used successfully across applications, species, and cell types. These include flow cytometry of endogenously expressing cells, real-time confocal microscopy of mouse splenocytes and human macrophages, as well as FRET-based kinetic and equilibrium binding assays. High CB2R specificity was demonstrated by competition experiments in living cells expressing CB2R at native levels. The probes were effectively applied to FACS analysis of microglial cells derived from a mouse model relevant to Alzheimer’s disease.
DOI: 10.1021/jacs.0c05587
Authors: Roman
C. Sarott, Matthias V. Westphal, Patrick Pfaff, Claudia Korn, David A. Sykes, Thais Gazzi, Benjamin Brennecke, Kenneth Atz, Marie Weise, Yelena Mostinski, Pattarin Hompluem, Eline Koers, Tamara Miljuš, Nicolas J. Roth, Hermon Asmelash, Man C. Vong, Jacopo Piovesan, Wolfgang Guba, Arne C. Rufer, Eric A. Kusznir, Sylwia Huber, Catarina Raposo, Elisabeth A. Zirwes, Anja Osterwald, Anto Pavlovic, Svenja Moes, Jennifer Beck, Irene Benito-Cuesta, Teresa Grande, Samuel Ruiz de Martı́n Esteban, Alexei Yeliseev, Faye Drawnel, Gabriella Widmer, Daniela Holzer, Tom van der Wel, Harpreet Mandhair, Cheng-Yin Yuan, William R. Drobyski, Yurii Saroz, Natasha Grimsey, Michael Honer, Jürgen Fingerle, Klaus Gawrisch, Julian Romero, Cecilia J. Hillard, Zoltan V. Varga, Mario van der Stelt, Pal Pacher, Jürg Gertsch, Peter J. McCormick, Christoph Ullmer, Sergio Oddi, Mauro Maccarrone, Dmitry B. Veprintsev, Marc Nazaré, Uwe Grether, Erick M. Carreira
Institutions: ETH Zurich, Laboratory of Organic Chemistry, F. Hoffmann-La Roche AG, University of Nottingham, University of Birmingham, Leibniz-Forschungsinstitut für Molekulare Pharmakologie, Queen Mary University of London, William Harvey Research Institute, Universidad Francisco de Vitoria, National Institutes of Health, National Institute on Alcohol Abuse and Alcoholism, Leiden University, University of Bern, Medical College of Wisconsin, University of Auckland, Semmelweis University, University of Teramo, Fondazione Santa Lucia, University of L'Aquila
Language: English
Subfield: Pharmacology
Field: Medicine
Domain: Health Sciences
Open Access status: green
Funders: National Science Foundation, Roche, F. Hoffmann-La Roche, University of Bern, Queen Mary University of London, Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung, Canada Excellence Research Chairs, Government of Canada, Eidgenössische Technische Hochschule Zürich, Ministerio de Economía y Competitividad, Ministero dell’Istruzione, dell’Università e della Ricerca, Università degli Studi dell'Aquila, Medical Research Council, Leibniz-Gemeinschaft, National Heart, Lung, and Blood Institute
Awards: Chemistry of Bioactive Agents, Structural and biophysical pharmacology of cannabinoid CB2 and vasopressin V2R G protein-coupled receptors, Genetic dissociation of GABA-B and gamma-hydroxybutyrate (GHB) receptor actions, G2241683054, Cannabinoid-mediated mitigation of graft versus host disease: Roles of CB2 receptors and adenosine signaling, G7134889682, Mechanistic Inflammatory Pathways in Graft Versus Host Disease, G8545618387
OpenAlex ID: W3083832329