title: High content screening for G protein-coupled receptors using cell-based protein translocation assays.
abstract: G protein-coupled receptors (GPCRs) have been one of the most productive classes of drug targets for several decades, and new technologies for GPCR-based discovery promise to keep this field active for years to come. While molecular screens for GPCR receptor agonist- and antagonist-based drugs will continue to be valuable discovery tools, the most exciting developments in the field involve cell-based assays for GPCR function. Some cell-based discovery strategies, such as the use of beta-arrestin as a surrogate marker for GPCR function, have already been reduced to practice, and have been used as valuable discovery tools for several years. The application of high content cell-based screening to GPCR discovery has opened up additional possibilities, such as direct tracking of GPCRs, G proteins and other signaling pathway components using intracellular translocation assays. These assays provide the capability to probe GPCR function at the cellular level with better resolution than has previously been possible, and offer practical strategies for more definitive selectivity evaluation and counter-screening in the early stages of drug discovery. The potential of cell-based translocation assays for GPCR discovery is described, and proof-of-concept data from a pilot screen with a CXCR4 assay are presented. This chemokine receptor is a highly relevant drug target which plays an important role in the pathogenesis of inflammatory disease and also has been shown to be a co-receptor for entry of HIV into cells as well as to play a role in metastasis of certain cancer cells.
authors.affiliation: BioImage A/S, Copenhagen, Denmark.
authors.forename: Charlotta
authors.initials: C
authors.lastname: Grånäs
authors.forename: Betina Kerstin
authors.initials: BK
authors.lastname: Lundholt
authors.forename: Arne
authors.initials: A
authors.lastname: Heydorn
authors.forename: Viggo
authors.initials: V
authors.lastname: Linde
authors.forename: Hans-Christian
authors.initials: HC
authors.lastname: Pedersen
authors.forename: Christian
authors.initials: C
authors.lastname: Krog-Jensen
authors.forename: Mette M
authors.initials: MM
authors.lastname: Rosenkilde
authors.forename: Len
authors.initials: L
authors.lastname: Pagliaro
mesh_terms: D000818:Animals; D001681:Biological Assay; D001859:Bone Neoplasms; D020126:Brefeldin A / Q000494:pharmacology; D019556:COS Cells; D054377:Chemokine CXCL12; D019743:Chemokines, CXC / Q000378:metabolism / Q000494:pharmacology; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D006056:Golgi Apparatus / Q000187:drug effects / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D012516:Osteosarcoma; D021381:Protein Transport; D019718:Receptors, CXCR4 / Q000819:agonists / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000819:agonists / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured
chemical_list: C515255:CXCL12 protein, human; D054377:Chemokine CXCL12; D019743:Chemokines, CXC; D019718:Receptors, CXCR4; D043562:Receptors, G-Protein-Coupled; D049452:Green Fluorescent Proteins; D020126:Brefeldin A