Human leukocytes differentially express endocannabinoid-glycerol lipases and hydrolyze 2-arachidonoyl-glycerol and its metabolites from the 15-lipoxygenase and cyclooxygenase pathways

©2019 Society for Leukocyte Biology..

2-Arachidonoyl-glycerol (2-AG) is an endocannabinoid with anti-inflammatory properties. Blocking 2-AG hydrolysis to enhance CB2 signaling has proven effective in mouse models of inflammation. However, the expression of 2-AG lipases has never been thoroughly investigated in human leukocytes. Herein, we investigated the expression of seven 2-AG hydrolases by human blood leukocytes and alveolar macrophages (AMs) and found the following protein expression pattern: monoacylglycerol (MAG lipase; eosinophils, AMs, monocytes), carboxylesterase (CES1; monocytes, AMs), palmitoyl-protein thioesterase (PPT1; AMs), α/β-hydrolase domain (ABHD6; mainly AMs), ABHD12 (all), ABHD16A (all), and LYPLA2 (lysophospholipase 2; monocytes, lymphocytes, AMs). We next found that all leukocytes could hydrolyze 2-AG and its metabolites derived from cyclooxygenase-2 (prostaglandin E2 -glycerol [PGE2 -G]) and the 15-lipoxygenase (15-hydroxy-eicosatetraenoyl-glycerol [15-HETE-G]). Neutrophils and eosinophils were consistently better at hydrolyzing 2-AG and its metabolites than monocytes and lymphocytes. Moreover, the efficacy of leukocytes to hydrolyze 2-AG and its metabolites was 2-AG  ≥ 15-HETE-G >> PGE2 -G for each leukocyte. Using the inhibitors methylarachidonoyl-fluorophosphonate (MAFP), 4-nitrophenyl-4-(dibenzo[d][1,3]dioxol-5-yl(hydroxy)methyl)piperidine-1-carboxylate (JZL184), Palmostatin B, 4'-carbamoylbiphenyl-4-yl methyl(3-(pyridin-4-yl)benzyl)carbamate, N-methyl-N-[[3-(4-pyridinyl)phenyl]methyl]-4'-(aminocarbonyl)[1,1'-biphenyl]-4-yl ester carbamic acid (WWL70), 4'-[[[methyl[[3-(4-pyridinyl)phenyl]methyl]amino]carbonyl]oxy]-[1,1'-biphenyl]-4-carboxylic acid, ethyl ester (WWL113), tetrahydrolipstatin, and ML349, we could not pinpoint a specific hydrolase responsible for the hydrolysis of 2-AG, PGE2 -G, and 15-HETE-G by these leukocytes. Furthermore, JZL184, a selective MAG lipase inhibitor, blocked the hydrolysis of 2-AG, PGE2 -G, and 15-HETE-G by neutrophils and the hydrolysis of PGE2 -G and 15-HETE-G by lymphocytes, two cell types with limited/no MAG lipase. Using an activity-based protein profiling (ABPP) probe to label hydrolases in leukocytes, we found that they express many MAFP-sensitive hydrolases and an unknown JZL184-sensitive hydrolase of ∼52 kDa. Altogether, our results indicate that human leukocytes are experts at hydrolyzing 2-AG and its metabolites via multiple lipases and probably via a yet-to-be characterized 52 kDa hydrolase. Blocking 2-AG hydrolysis in humans will likely abrogate the ability of human leukocytes to degrade 2-AG and its metabolites and increase their anti-inflammatory effects in vivo.

Medienart:	E-Artikel

Erscheinungsjahr:	2019
Erschienen:	2019

Enthalten in:	Zur Gesamtaufnahme - volume:106
Enthalten in:	Journal of leukocyte biology - 106(2019), 6 vom: 02. Dez., Seite 1337-1347

Sprache:	Englisch

Beteiligte Personen:	Turcotte, Caroline [VerfasserIn] Dumais, Élizabeth [VerfasserIn] Archambault, Anne-Sophie [VerfasserIn] Martin, Cyril [VerfasserIn] Blanchet, Marie-Renée [VerfasserIn] Bissonnette, Élyse [VerfasserIn] Boulet, Louis-Philippe [VerfasserIn] Laviolette, Michel [VerfasserIn] Di Marzo, Vincenzo [VerfasserIn] Flamand, Nicolas [VerfasserIn]

Links:	Volltext

Themen:	15-lipoxygenase 2-arachidonoyl-glycerol 8D239QDW64 Arachidonate 15-Lipoxygenase Arachidonic Acids Biomarkers EC 1.13.11.33 EC 1.14.99.1 Endocannabinoid Endocannabinoids Enzyme Inhibitors Glycerides Glyceryl 2-arachidonate Journal Article Prostaglandin Prostaglandin-Endoperoxide Synthases Research Support, Non-U.S. Gov't

Anmerkungen:	Date Completed 03.06.2020 Date Revised 03.06.2020 published: Print-Electronic Citation Status MEDLINE

doi:	10.1002/JLB.3A0919-049RRR

funding:
Förderinstitution / Projekttitel:

PPN (Katalog-ID):	NLM30164201X