INTERGLU · Dissecting mGlu5 receptor internalization pathways using genetic and pharmacological tools
Horizon 2020 — Marie Skłodowska-Curie Actions
- Duration
- 2018-11-01 → 2020-10-31
- EU contribution
- €212,195
- Participants
- 1
- Scheme
- MSCA-IF-EF-ST
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Results in brief
Dissecting mGlu5 receptor internalization pathways using genetic and pharmacological tools
G protein-coupled receptors (GPCRs) are membrane proteins that are activated by a broad range of ligands and among the most abundant gene families in the human genome. GPCRs are involved in many important physiological processes and are the target of 30-40% of all marketed drugs. The most important function of GPCRs is to activate intracellular signaling cascades that regulate cellular function. Activation of intracellular partners is regulated by receptor internalization to avoid overstimulation of the cell. The metabotropic glutamate receptor 5 (mGlu5) is a class C GPCR that plays a key role in synaptic plasticity, generally thought to be the basis of learning and memory formation. Thus, it is not surprising that mGlu5 is a promising drug target for several central nervous system disorders, including Fragile X syndrome, Parkinson’s disease, addictive disorders, anxiety and schizophrenia. The receptor internalization mechanism has been well-characterized for a few GPCRs and a generalized model has been developed. However, the receptor internalization pathway has not been studied for the majority of GPCRs and it remains unclear whether they follow the canonical pathway. In fact, there is an increasing number of studies suggesting that non-canonical receptor internalization is more prevalent than previously anticipated. The overall objectives of the INTERGLU project (grant agreement number 797497) were: (1) to develop novel genetic and pharmacological tools to study GPCR internalization and (2) to apply them to dissect the internalization mechanisms of GPCRs and in particular the mGlu5 receptor.
Data: CORDIS, © European Union
Project objective
The metabotropic glutamate receptor 5 (mGlu5) is part of the large family of G protein-coupled receptors (GPCRs) that are the target of 30-40% of marketed drugs. mGlu5 is a key regulator of synaptic plasticity in the brain and a major drug target for a wide range of diseases, such as Fragile X syndrome. Agonist-induced internalization is a major regulatory mechanism for GPCR signalling. Interestingly, mGlu5 internalizes via a mechanism that is distinct from the canonical beta-arrestin mediated pathway. However, the details of this alternative mechanism are largely unknown for mGlu5 and poorly understood for GPCRs in general. The present project has the dual aim (1) to dissect the intracellular pathways leading to mGlu5 receptor internalization and its functional implications and (2) to develop a genetic and pharmacological toolbox to study beta-arrestin independent internalization.I will generate a series of HEK293 cells where key proteins involved in a range of GPCR internalization pathways have been systematically removed by CRISPR/Cas9 genome editing. These cell lines will complement the already existing G protein alpha subunit and beta-arrestin knockout HEK293 cell lines to generate a highly efficient toolbox to study pathways and mechanisms involved in GPCR signalling, internalization and regulation. In the present project, I will employ theses cell lines to dissect pathways leading to mGlu5 internalization. To enable future studies in cells with native mGlu5 receptor expression, pharmacological tool compounds/peptides targeting key proteins identified will be developed using the genome edited HEK293 cells. This will provide essential information about the regulation of this important drug target and beta-arrestin independent internalization.Importantly, we will make the cell lines and pharmacological tool compounds available to the scientific community enabling similar studies on a much wider range of GPCRs.
Original text from CORDIS.
Participants
- KOBENHAVNS UNIVERSITET · KOBENHAVNCoordinatorDenmark
Links
- View on CORDIS
- DOI: 10.3030/797497
- https://drug.ku.dk/disciplines/molecular-and-cellular-pharmacology/molecular-pharmacology-of-g-protein-coupled-receptors/
Data: CORDIS, © European Union