A broad-spectrum lipidomics screen of antiinflammatory drug combinations in human blood

Abstract

Current methods of drug screening in human blood focus on the immediate products of the affected pathway and mostly rely on approaches that lack sensitivity and the capacity for multiplex analysis. We have developed a sensitive and selective method based on ultra-performance liquid chromatography–tandem mass spectrometry to scan the effect of drugs on the bioactive eicosanoid lipidome in vitro and ex vivo. Using small sample sizes, we can reproducibly measure a broad spectrum of eicosanoids in human blood and capture drug-induced substrate rediversion and unexpected shifts in product formation. Microsomal prostaglandin E synthase-1 (mPGES-1) is an antiinflammatory drug target alternative to COX-1/-2. Contrasting effects of targeting mPGES-1 versus COX-1/-2, due to differential substrate shifts across the lipidome, were observed and can be used to rationalize and evaluate drug combinations. Finally, the in vitro results were extrapolated to ex vivo studies by administration of the COX-2 inhibitor, celecoxib, to volunteers, illustrating how this approach can be used to integrate preclinical and clinical studies during drug development.

Targeting biosynthetic networks of the proinflammatory and proresolving lipid metabolome

“…In both macrophage phenotypes, particularly in M1, the biosynthesis of the 5-LOX products LTB 4 and its trans-isomers, 5-HETE, 5-HEPE, and 5S,6R-diHETE was strongly increased by celecoxib and, to a minor degree, by ibuprofen (at least in M1, Fig. 2), probably as a result of AA substrate shunting from the COX toward the 5-LOX and FLAP pathway as reported by Mazaleuskaya et al (26). Notably, also 7-HDHA levels were increased by celecoxib in M1 ( Fig.…”

“…LMs produced via the COX‐1/2 pathway display proinflammatory but also protective actions depending on stimulus, cell type, and status that program the ability to resolve inflammation (5, 38). Although beneficial effects of NSAIDs are well established for alleviating acute inflammation and pain (26, 39), they cause severe on‐target side effects and may negatively impact inflammation resolution (35, 40). Thus, COX‐2–deficient mice failed to resolve from inflammation, and the COX inhibitor indomethacin exacerbated inflammation because of reduced proresolving PGD 2 levels (41).…”

Flipping the cyclooxygenase (Ptgs) genes reveals isoform-specific compensatory functions ,

“…The lipidomics analysis of mouse plasma and serum was performed as described previously for human plasma (35) plasma and serum samples (20 l) in 300 l of acetonitrile. Then the samples were acidified by addition of formic acid to a final concentration of 1% and incubated at room temperature for 15 min.…”

MS-based lipidomics of human blood plasma: a community-initiated position paper to develop accepted guidelines

“…We suggest that the minimum set of personal and clinical data collected along with plasma/serum samples should be subject age, gender, BMI, ethnicity, fasting status, and prescription medications, including drugs directly affecting lipid metabolism (e.g., nonsteroidal anti-inflammatory drugs, anticoagulants, and statins) and also drugs with insufficiently characterized metabolic impact (i.e., hormones, including contraceptives, steroids, and diuretics) ( 17 , 50 – 52 ). It should also include significant medical conditions (e.g., affected with chronic disease).…”