Chimeric GPCRs mimic distinct signaling pathways and modulate microglia responses
Ontology highlight
ABSTRACT: G protein-coupled receptors (GPCRs) regulate processes ranging from immune responses to neuronal signaling. However, ligands for many GPCRs remain unknown, suffer from off-target effects or have poor bioavailability. Additionally, dissecting cell type-specific responses is challenging when the same GPCR is expressed on different cells within a tissue. Here, we overcome these limitations by engineering DREADD-based GPCR chimeras that bind their agonist clozapine-N-oxide (CNO) and mimic a GPCR-of-interest. We show that the chimeric DREADD-β2AR triggers responses comparable to levalbuterol-stimulated β2AR on second messenger and kinase activity, post-translational modifications, and protein-protein interactions. Moreover, we successfully recapitulate β2AR-mediated filopodia formation in primary microglia, an immune cell capable of driving central nervous system inflammation. When dissecting microglial inflammation, we utilized the microglia-like HMC3 cell line and included two additionally designed DREADD-based chimeras mimicking GPR65 and GPR109A. DREADD-β2AR and DREADD-GPR65 modulate the inflammatory response with high similarity to endogenous β2AR, while DREADD-GPR109A had no impact. Our DREADD-based approach allows investigation of cell type-dependent pathways without known endogenous ligands.
ORGANISM(S): Homo sapiens
PROVIDER: GSE194125 | GEO | 2022/07/06
REPOSITORIES: GEO
ACCESS DATA