Molecular & Translational Drug Discovery
Single-particle cryo-EM workflow for high resolution structure-guided drug discovery
Molecular and Cellular Pharmacology
To probe molecular mechanisms of action, we utilise a range of biochemical, pharmacological and cell-signalling assays including radioligand binding, TR-FRET, calcium mobilisation, cAMP accumulation, GTPγ35S binding, ERK1/2 phosphorylation and label-free technology. Additionally, we develop and run cellular assays in 384-well plate format to enable high-throughput and primary SAR screening.
Kinetic and selectivity differentiation of histamine H3 receptor antagonists. Adapted from Riddy et al., (2019) Neuropharmacology (doi:10.1016/j.neuropharm.2018.10.028)
Native and Human Tissue Pharmacology
To better reflect the in vivo situation, we study GPCR expression and pharmacology (using many of the assays above) in native tissue and cellular systems, including isolated human and rodent brain tissue, primary cultured neurons, and isolated primary blood cells.
Expression profiling of the orphan receptor, Gpr52 (white), in the dorsal striatum (DS) of mouse brain using RNAscope fluorescent in situ hybridisation. Gpr52 is primarily co-expressed with the dopamine D2 receptor (Drd2; red), but not the dopamine D1 receptor (Drd1; green), in medium spiny neurons (adapted from Spark et al., 2020 [doi.org/10.1021/acschemneuro.0c00197]).
Atherosclerotic plaque from the aorta of a wild-type mouse fed a 60% high fat diet for 20 weeks, as imaged using Coherent Anti-Stokes Raman Scattering (CARS) microscopy (white = macrophages / lipids; green = collagen; Sanja Bosnyak-Gladovic & Cameron Nowell).
To provide translation from recombinant and cellular studies to in vivo models and better understand drug mechanism of action, we utilise a range of translational approaches, including in vivo neurobiology, obesity and metabolism models, and patient-derived blood samples.
Phenotype of wild-type and orphan GPCR knockout mice in the object-location paired associates learning (PAL) touchscreen task (Stewart et al., 2020 [unpublished observations]).
Microinjection of the muscarinic M5 receptor negative allosteric modulator (NAM), ML375, in the dorsolateral, but nor dorsomedial striatum of alcohol-experienced iP rats reduces alcohol self-administration (adapted from Berizzi et al., (2018) Neuropsychopharmacology; doi 10.1038/s41386-017-0007-3).
For further information, please contact Chris Langmead.