Detection of cannabinoid receptor type 2 in native cells and zebrafish with a highly potent, cell-permeable fluorescent probe

Gazzi, Thais; Brennecke, Benjamin; Atz, Kenneth; Korn, Claudia; Sykes, David; Forn-Cuni, Gabriel; Pfaff, Patrick; Sarott, Roman C.; Westphal, Matthias V.; Mostinski, Yelena; Mach, Leonard; Wasinska-Kalwa, Malgorzata; Weise, Marie; Hoare, Bradley L.; Miljuš, Tamara; Mexi, Maira; Roth, Nicolas; Koers, Eline J.; Guba, Wolfgang; Alker, André; Rufer, Arne C.; Kusznir, Eric A.; Huber, Sylwia; Raposo, Catarina; Zirwes, Elisabeth A.; Osterwald, Anja; Pavlovic, Anto; Moes, Svenja; Beck, Jennifer; Nettekoven, Matthias; Benito-Cuesta, Irene; Grande, Teresa; Drawnel, Faye; Widmer, Gabriella; Holzer, Daniela; van der Wel, Tom; Mandhair, Harpreet; Honer, Michael; Fingerle, Jürgen; Scheffel, Jörg; Broichhagen, Johannes; Gawrisch, Klaus; Romero, Julián; Hillard, Cecilia J.; Varga, Zoltan V.; van der Stelt, Mario; Pacher, Pal; Gertsch, Jürg; Ullmer, Christoph; McCormick, Peter J.; Oddi, Sergio; Spaink, Herman P.; Maccarrone, Mauro; Veprintsev, Dmitry B.; Carreira, Erick M.; Grether, Uwe; Nazaré, Marc

Abstract

Despite its essential role in the (patho)physiology of several diseases, CB2R tissue expression profiles and signaling mechanisms are not yet fully understood. We report the development of a highly potent, fluorescent CB2R agonist probe employing structure-based reverse design. It commences with a highly potent, preclinically validated ligand, which is conjugated to a silicon-rhodamine fluorophore, enabling cell permeability. The probe is the first to preserve interspecies affinity and selectivity for both mouse and human CB2R. Extensive cross-validation (FACS, TR-FRET and confocal microscopy) set the stage for CB2R detection in endogenously expressing living cells along with zebrafish larvae. Together, these findings will benefit clinical translatability of CB2R based drugs.

Citation

Gazzi, T., Brennecke, B., Atz, K., Korn, C., Sykes, D., Forn-Cuni, G., Pfaff, P., Sarott, R. C., Westphal, M. V., Mostinski, Y., Mach, L., Wasinska-Kalwa, M., Weise, M., Hoare, B. L., Miljuš, T., Mexi, M., Roth, N., Koers, E. J., Guba, W., Alker, A., …Nazaré, M. (2022). Detection of cannabinoid receptor type 2 in native cells and zebrafish with a highly potent, cell-permeable fluorescent probe. Chemical Science, 13(19), 5539-5545. https://doi.org/10.1039/d1sc06659e

• Journal Article Type: Article
• Acceptance Date: Jan 22, 2022
• Online Publication Date: Apr 1, 2022
• Publication Date: May 21, 2022
• Deposit Date: Jun 20, 2025
• Publicly Available Date: Jun 23, 2025
• Journal: Chemical Science
• Print ISSN: 2041-6520
• Electronic ISSN: 2041-6539
• Publisher: Royal Society of Chemistry
• Peer Reviewed: Peer Reviewed
• Volume: 13
• Issue: 19
• Pages: 5539-5545
• DOI: https://doi.org/10.1039/d1sc06659e
• Public URL: https://nottingham-repository.worktribe.com/output/8956014
• Publisher URL: https://pubs.rsc.org/en/content/articlelanding/2022/sc/d1sc06659e