title: DAGLβ inhibition perturbs a lipid network involved in macrophage inflammatory responses.
abstract: The endocannabinoid 2-arachidonoylglycerol (2-AG) is biosynthesized by diacylglycerol lipases DAGLα and DAGLβ. Chemical probes to perturb DAGLs are needed to characterize endocannabinoid function in biological processes. Here we report a series of 1,2,3-triazole urea inhibitors, along with paired negative-control and activity-based probes, for the functional analysis of DAGLβ in living systems. Optimized inhibitors showed high selectivity for DAGLβ over other serine hydrolases, including DAGLα (∼60-fold selectivity), and the limited off-targets, such as ABHD6, were also inhibited by the negative-control probe. Using these agents and Daglb(-/-) mice, we show that DAGLβ inactivation lowers 2-AG, as well as arachidonic acid and eicosanoids, in mouse peritoneal macrophages in a manner that is distinct and complementary to disruption of cytosolic phospholipase-A2. We observed a corresponding reduction in lipopolysaccharide-induced tumor necrosis factor-α release. These findings indicate that DAGLβ is a key metabolic hub within a lipid network that regulates proinflammatory responses in macrophages.
authors.affiliation: The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, California, USA.
authors.forename: Ku-Lung
authors.initials: KL
authors.lastname: Hsu
authors.forename: Katsunori
authors.initials: K
authors.lastname: Tsuboi
authors.forename: Alexander
authors.initials: A
authors.lastname: Adibekian
authors.forename: Holly
authors.initials: H
authors.lastname: Pugh
authors.forename: Kim
authors.initials: K
authors.lastname: Masuda
authors.forename: Benjamin F
authors.initials: BF
authors.lastname: Cravatt
mesh_terms: D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism; D001095:Arachidonic Acids / Q000096:biosynthesis; D002460:Cell Line; D016207:Cytokines / Q000378:metabolism; D055808:Drug Discovery; D063388:Endocannabinoids / Q000096:biosynthesis; D005989:Glycerides / Q000096:biosynthesis; D007249:Inflammation / Q000378:metabolism*; D050356:Lipid Metabolism / Q000187:drug effects*; D008071:Lipoprotein Lipase / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000502:physiology; D017737:Macrophages, Peritoneal / Q000187:drug effects* / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D011453:Prostaglandins / Q000378:metabolism; D020033:Protein Isoforms; D020543:Proteome / Q000187:drug effects; D021281:Quantitative Structure-Activity Relationship; D015398:Signal Transduction / Q000187:drug effects; D014230:Triazoles / Q000138:chemical synthesis / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D001095:Arachidonic Acids; D016207:Cytokines; D063388:Endocannabinoids; D005989:Glycerides; D011453:Prostaglandins; D020033:Protein Isoforms; D020543:Proteome; D014230:Triazoles; D014409:Tumor Necrosis Factor-alpha; D016718:Arachidonic Acid; C094503:glyceryl 2-arachidonate; D008071:Lipoprotein Lipase; C577904:diacylglycerol lipase beta, mouse