Myeloid-specific deficiency of long-chain acyl CoA synthetase 4 (Acsl4) reduces inflammation in vitro and in vivo by remodeling phospholipids and reducing production of arachidonic acid-derived proinflammatory lipid mediators

In response to infection or tissue damage, resident peritoneal macrophages (rpMACs) produce inflammatory lipid mediators from the polyunsaturated fatty acid (PUFA), arachidonic acid (AA). Long-chain acyl-CoA synthetase 4 (ACSL4) catalyzes the covalent addition of a CoA moiety to fatty acids, with a strong preference for AA and other PUFA containing three or more double bonds. PUFA-CoA can be incorporated into phospholipids, which is the source of PUFA for lipid mediator synthesis. In this study, we demonstrated that deficiency of Acsl4 in mouse rpMACs resulted in a significant reduction of AA incorporated into all phospholipid classes and a reciprocal increase in incorporation of oleic acid and linoleic acid. After stimulation with opsonized zymosan (opZym) a diverse array of AA-derived lipid mediators, including leukotrienes, prostaglandins, HETEs, and lipoxins, were produced and were significantly reduced in Acsl4 -deficient rpMACs. The Acsl4 -deficient rpMACs stimulated with opZym also demonstrated an acute reduction in mRNA expression of the inflammatory cytokines, Il6 , Ccl2 , Nos2 and Ccl5 . When Acsl4 -deficient rpMACS were incubated in vitro with the TLR4 agonist, LPS, the levels of leukotriene B 4 (LTB 4 ) and prostaglandin E 2 (PGE 2 ) were also significantly decreased. In LPS-induced peritonitis, mice with myeloid-specific Acsl4 deficiency had a significant reduction in LTB 4 and PGE 2 levels in peritoneal exudates, which was coupled with reduced infiltration of neutrophils in the peritoneal cavity as compared with WT mice. Our data demonstrate that chronic deficiency of Acsl4 in rpMACs reduces the incorporation of AA into phospholipids, which reduces lipid mediator synthesis and inflammation.