DAGL[beta] inhibition perturbs a lipid network involved in macrophage inflammatory responses

• Published in: Nature chemical biology Vol. 8; no. 12; p. 999
• Main Authors: Hsu, Ku-lung, Tsuboi, Katsunori, Adibekian, Alexander, Pugh, Holly, Masuda, Kim, Cravatt, Benjamin F
• Format: Journal Article
• Language: English
• Published: Cambridge Nature Publishing Group 01.12.2012
• Subjects: Biosynthesis; Enzymes; Inactivation; Inhibitors; Lipids; Molecular chemistry; Probes; Rodents; Urea
• ISSN: 1552-4450; 1552-4469
• DOI: 10.1038/nchembio.1105

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. [PUBLICATION ABSTRACT]