Highly multiplexed bioactivity screening reveals human and microbiota metabolome-GPCRome interactions

• Published in: Cell Vol. 186; no. 14; pp. 3095 - 3110.e19
• Main Authors: Chen, Haiwei, Rosen, Connor E., González-Hernández, Jaime A., Song, Deguang, Potempa, Jan, Ring, Aaron M., Palm, Noah W.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 06.07.2023
• Subjects: Animals; bioactive properties; diet; drugs; G protein-coupled receptor (GPCR); gingipain; high-throughput screening; human metabolome; Humans; landscapes; Mammals - metabolism; metabolites; Metabolome; microbiome; Microbiota; microbiota metabolites; microorganisms; Porphyromonas gingivalis; Receptors, G-Protein-Coupled - metabolism; G protein-coupled receptor (GPCR); microbiome; microbiota metabolites; human metabolome; high-throughput screening
• ISSN: 0092-8674; 1097-4172; 1097-4172
• DOI: 10.1016/j.cell.2023.05.024

The human body contains thousands of metabolites derived from mammalian cells, the microbiota, food, and medical drugs. Many bioactive metabolites act through the engagement of G-protein-coupled receptors (GPCRs); however, technological limitations constrain current explorations of metabolite-GPCR interactions. Here, we developed a highly multiplexed screening technology called PRESTO-Salsa that enables simultaneous assessment of nearly all conventional GPCRs (>300 receptors) in a single well of a 96-well plate. Using PRESTO-Salsa, we screened 1,041 human-associated metabolites against the GPCRome and uncovered previously unreported endogenous, exogenous, and microbial GPCR agonists. Next, we leveraged PRESTO-Salsa to generate an atlas of microbiome-GPCR interactions across 435 human microbiome strains from multiple body sites, revealing conserved patterns of cross-tissue GPCR engagement and activation of CD97/ADGRE5 by the Porphyromonas gingivalis protease gingipain K. These studies thus establish a highly multiplexed bioactivity screening technology and expose a diverse landscape of human, diet, drug, and microbiota metabolome-GPCRome interactions. [Display omitted] •PRESTO-Salsa enables multiplexed bioactivity screening of >300 GPCRs via NGS•Screening of human metabolome- and microbiota secretome-GPCRome interactions•Identification of cross-tissue human microbiome-GPCRome interactions•Porphyromonas gingivalis activates CD97/ADGRE5 via site-specific proteolytic cleavage PRESTO-Salsa is a multiplexed GPCR screening platform that enables the interrogation of GPCRome-wide bioactivities across diverse sample types. This platform was used to explore microbiota-induced GPCR activation and revealed the engagement of an immune cell-associated GPCR by a periodontal pathogen protease.