Investigating DAGLβ expression and activity in human monocytes via proteomic and lipidomic profiling

Abstract Lipid-metabolizing enzymes are important targets for small-molecule inhibitor discovery as the regulation of inflammation responses have been linked to lipid metabolism cascades controlled by these enzymes. Non-steroidal anti-inflammatory drugs inhibit in COX pathway preventing conversion arachidonic acid (AA) into inflammation-inducing prostaglandins. An upstream precursor AA is endocannabinoid 2-arachidonoyl-glycerol (2-AG), which biosynthesized from cleavage arachidonate-containing diacylglycerols (DAGs) diacylglycerol lipase beta (DAGLβ). Previous work identify DAGLβ’s role inflammatory function led creation KT109, a DAGLβ-specific 1,2,3-triazole urea inhibitor. Primary mouse macrophages treated with KT109 showed reductions and 2-AG an increase DAGs, demonstrating that DAGLβ key proinflammatory signaling. These studies focused on effects inhibiting DAGLβ, but information regarding inhibition human still lacking. Herein, we applied activity-based protein profiling (ABPP) quantitative proteomic lipidomic methods determine effectivity monocytic cell line THP1. Potency (IC 50values) THP1 cells were comparable magnitude cells. Lipidomic analysis KT109-treated macrophage-differentiated THP1s did not demonstrate accumulation DAGs. findings show potential difference expression activity when compared additional Funding disclosures: This was supported National Institute Drug Abuse grant no. R01 DA043571.