TY - JOUR T1 - High-throughput screening of TRPV1 ligands in the light of the Bioluminescence Resonance Energy Transfer technique JF - Molecular Pharmacology JO - Mol Pharmacol DO - 10.1124/molpharm.121.000271 SP - MOLPHARM-AR-2021-000271 AU - Yann Chappe AU - Pauline Michel AU - Alexandre Joushomme AU - Solène Barbeau AU - Sandra Pierredon AU - Luc Baron AU - André Garenne AU - Florence Poulletier De Gannes AU - Annabelle Hurtier AU - Stanislas Mayer AU - Isabelle Lagroye AU - Jean-François Quignard AU - Thomas Ducret AU - Vincent Compan AU - Christelle Franchet AU - Yann Percherancier Y1 - 2021/01/01 UR - http://molpharm.aspetjournals.org/content/early/2021/06/12/molpharm.121.000271.abstract N2 - Ion channels are attractive drug targets for many therapeutic applications. However, high-throughput screening (HTS) of drug candidates is difficult and remains very expensive. We thus assessed the suitability of the Bioluminescence Resonance Energy Transfer (BRET) technique as a new HTS method for ion-channel studies by taking advantage of our recently characterized intra- and intermolecular BRET probes targeting the TRPV1 ion channel. These BRET probes monitor conformational changes during TRPV1 gating and subsequent coupling with Calmodulin, two molecular events that are intractable using reference techniques such as automated calcium assay (ACA) and automated patch-clamp (APC). We screened the small-sized Prestwick chemical library, encompassing 1200 compounds with high structural diversity, using either intra- and intermolecular BRET probes or ACA. Secondary screening of the detected hits was done using APC. Multiparametric analysis of our results shed light on the capability of calmodulin inhibitors included in the Prestwick library to inhibit TRPV1 activation by Capsaicin (CAPS). BRET was the lead technique for this identification process. Finally, we present data exemplifying the use of intramolecular BRET probes to study other TRPs and non-TRPs ion channels. Knowing the ease of use of BRET biosensors and the low cost of the BRET technique, these assays may advantageously be included for extending ion-channel drug screening. Significance Statement We screened a chemical library against TRPV1 ion channel using Bioluminescence Resonance Energy Transfer (BRET) molecular probes, and compared the results with the ones obtained using reference techniques such as automated calcium assay and automated patch-clamp. Multiparametric analysis of our results shed light on the capability of Calmodulin antagonists to inhibit chemical activation of TRPV1, and indicates that BRET probes may advantageously be included in ion channel drug screening campaigns. ER -