Nomad Biosensors: A New Multiplexed Technology for the Screening of GPCR Ligands

Abstract

Nomad Technology (Innoprot [Innovative Technologies in Biological Systems], Derio, Spain), a novel tool for multiplexing high-throughput cell-based G protein-coupled receptor (GPCR) assays, is described in this work. This new technology comprises a family of fluorescent biosensors called Nomad Biosensors that allow for the measurement of responses mediated by G proteins through their interactions with second-messenger transduction proteins. GPCRs are one of the largest protein families of receptors in eukaryotes, and their signaling mediates important physiological processes within cells. Thus, GPCRs are associated with a wide variety of diseases, and considered major targets in therapeutic research. Nomad constitutes a novel tool for unraveling the mechanism of GPCR signal transduction by simultaneously tracing different pathways. GPCR activation changes the structural folding of the biosensor and promotes its vesicularization, as well as an increase in the fluorescence intensity. Based on this technology, the Nomad cellular model was developed to discriminate between the Ca-mediated pathway and the cyclic adenosine monophosphate (cAMP)-mediated pathway. To validate this model, endothelin receptor B (ETR) was coexpressed into the Nomad cell line and assessed with a specific agonist, an antagonist, and a chemical library of compounds. Nomad Technology optimizes the identification of novel GPCR ligands and enables the testing of large numbers of compounds.

Protein-protein interaction modulators: advances, successes and remaining challenges

“…These new technologies, combined with cellular screening assays (including high-content screening), help to discover and design PPI modulators. Cellular assays help to assess the activity of whole intracellular pathways or portions of pathways to identify compounds that inhibit these pathways via disruption of key PPIs (Mella et al 2018;Booij et al 2019). Two examples of cellular screening assays are ligand signal transducers and activators of transcription (STATs) technology and BioImage Redistribution technology.…”

Δ9‐Tetrahydrocannabinolic acid alleviates collagen‐induced arthritis: Role of PPARγ and CB1 receptors

“…Based on Nomad Biosensor technology, the cell line was created to identify β‐arrestin recruitment following CB 1 receptor activation. To accomplish this, β‐arrestin was coupled to reporters engineered with a single fluorescence protein that transduce the signal upon ligand binding to CB 1 receptors (Mella et al, 2018). Thus, in HEK293‐CB 1 ‐β‐arrestin Nomad cells, the green β‐arrestin biosensor become fluorescent upon activation of CB 1 receptors.…”

Quantitative confocal microscopy for grouping of dose–response data: Deciphering calcium sequestration and subsequent cell death in presence of excess norepinephrine

“…Additionally, Fluo-4 has high dynamic range, brightness, and high signalto-noise ration among various Ca 2+ dyes. 16 It is known that 34% of drugs on the market are based on G protein-coupled receptor (GPCR)-targeting drugs 17,18 that are used to treat chronic heart disease, neurodisease, and cancer. It has also been shown that Ca 2+ imaging can be used for drug screening and to identify the operating range of GPCR-targeting drug doses.…”