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Pharmacological Perturbations of Eicosanoids in vitro and ex vivo in a Novel, Broad‐spectrum, Plasma Lipidomics Assay
Author(s) -
Mazaleuskaya Liudmila L,
Ricciotti Emanuela,
Lawson John A,
Li Xuanwen,
Grant Gregory,
Grosser Tilo,
FitzGerald Garret A
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.1198.1
Subject(s) - ex vivo , lipidomics , cyclooxygenase , prostacyclin , pharmacology , eicosanoid , prostaglandin , in vivo , chemistry , arachidonic acid , lipid signaling , prostaglandin e2 , arachidonate 5 lipoxygenase , in vitro , biochemistry , medicine , enzyme , biology , microbiology and biotechnology
Eicosanoids are a family of bioactive lipids formed from arachidonic acid and include prostaglandins, leukotrienes and related compounds. Nonsteroidal anti‐inflammatory drugs (NSAIDs) suppress prostaglandins and are widely used for anti‐pyretic, analgesic and anti‐inflammatory effects. NSAIDs selective for inhibition of cyclooxygenase (COX)‐2 have fewer gastro‐intestinal side effects, but predispose patients to increased cardiovascular risk. The cardiovascular complications prompted interest in microsomal prostaglandin synthase 1 (mPGES‐1), downstream of COX‐2, as a novel drug target. Unlike COX‐2, global deletion of mPGES‐1 in mice suppresses prostaglandin E 2 (PGE 2 ) and augments prostacyclin (PGI 2 ) through substrate rediversion, and does not predispose to thrombogenesis and hypertension. However, cell‐specific deletions of mPGES‐1 reveal that the predominant prostaglandin product of substrate rediversion varies by cell type. We hypothesized that inhibition of mPGES‐1 leads to changes in bioactive lipids beyond the prostaglandin pathway and aimed 1) to develop a mass spectrometry‐based, broad‐spectrum lipidomics approach to measure arachidonate‐derived products additional to prostanoids and 2) to develop a human whole blood assay (hWBA) to study substrate rediversion upon mPGES‐1 vs COX‐2 inhibition. We have developed a novel, reproducible, analytical method to scan the impact of drugs on the bioactive lipidome in human whole blood in vitro and ex vivo . We observed substrate rediversion in both COX and LOX (lipoxygenase) pathways after mPGES‐1 inhibition, which was distinct from inhibiting COX‐1/2. We validated our in vitro assay by its application ex vivo after administration of a COX‐2 inhibitor to human subjects. Overall, we have shown that blockade, alone and in combination, of diverse targets in the eicosanoid synthesis/response cascade results in both expected and unanticipated signals in the circulating human lipidome. Such novel aspects of drug action prompt reconsideration of how drugs in these pathways might be utilized in the alleviation of disease. Support or Funding Information This study was supported by The Personalized NSAID Therapeutics Consortium (PENTACON: HL117798) and by T32 H107971.