MS-based targeted metabolomics of eicosanoids and other oxylipins: Analytical and inter-individual variabilities Cécile Gladine, A. I. Ostermann, J. W. Newman, N. H. Schebb To cite this version: Cécile Gladine, A. I. Ostermann, J. W. Newman, N. H. Schebb. MS-based targeted metabolomics of eicosanoids and other oxylipins: Analytical and inter-individual variabilities. Free Radical Biology and Medicine, Elsevier, 2019, 144 (SI), 10.1016/j.freeradbiomed.2019.05.012. hal-02154450 HAL Id: hal-02154450 https://hal.archives-ouvertes.fr/hal-02154450 Submitted on 3 Dec 2019 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. ACCEPTED MANUSCRIPT 1 MS-based targeted metabolomics of eicosanoids and other oxylipins: analytical and inter- 2 individual variabilities. 3 4 Cécile Gladine 1*, Annika I. Ostermann 2, John W Newman 3,4 , Nils Helge Schebb 2 5 1Université Clermont Auvergne, INRA, UNH, Unité de Nutrition Humaine, CRNH Auvergne, Clermont- 6 Ferrand, France. 7 2 Chair of Food Chemistry, Faculty of Mathematics and Natural Sciences, Gaußstraße 20, University of 8 Wuppertal, 42119, Wuppertal Germany 9 3 United States Department of Agriculture, Agricultural Researh Service, Western Human Nutrition 10 Research Center, Davis CA, USA. 11 4 University of California Davis, Department of Nutrition, Davis CA, USA 12 *Correspondance : Dr Cécile Gladine, [email protected] , +33 4 73 62 42 30 13 1 1 Abbreviations : (U)HPLC-MS, (ultra) high performance liquid chromatography-mass spectrometry ; AA, arachidonic acid ; PUFAs, polyunsaturated fatty acids ; COX, cycloxygenase ; PG, prostaglandin ; Tx, thromboxanes ; LOX, lipoxygenase ; LT, leukotrienes ; CYP, cytochrome P450 ; EPA, eicosapentaenoic acid ; GPCR, G protein-coupled receptor ; TRPs, transient receptor potential channels ; PPARs, peroxisome proliferator-activated receptors ; NF κB, nuclear factor kappa B ; DHA, docosahexaenoic acid ; EpOME, epoxyoctadecenoicACCEPTED acid ; HODE, hydroxyoctadecadienoic acid; KODE, ketooctadecadienoic acid ; CRP, C- reactive protein ; HETE, hydroxyeicosatetraenoic acid ; HEPE, hydroxyeicosapentaenoic acid ; HDHA, hydroxydocosahexaenoic acid ; EDTA, ethylenediaminetetraacetic acid ; TriHOME, trihydroxyoctadecenoic acid ; BHT, butylated hydroxytoluene ; SPE, solid phase extraction ; ESI-MS, electrospray ionization-mass spectrometry ; GLC-MS, gas liquid chromatography mass spectrometry ; QqQ-MS, triple quadrupole mass spectrometry ; TOF, time of flight ; EpETrE, epoxyeicosatrienoic acid ; SPM, specialized pro-resolving mediators ; LX, lipoxin ; MaR, maresin ; PD, protectin ; LLOQ, lower limit of quantification ; LOD, limit of detection ; HMG- CoA, 3-Hydroxy-3-Methyl-Glutaryl-CoA reductase ; FLAP, 5-lipoxygenase-activating protein ; CVD, cardiovascular diseases ; sEH, soluble epoxyhydroalse ; CAD, coronary artery diseases. 1 ACCEPTED MANUSCRIPT 14 Abstract (150 words). 15 Oxylipins, including the well-known eicosanoids, are potent lipid mediators involved in numerous 16 physiological and pathological processes. Therefore, their quantitative profiling has gained a lot of 17 attention during the last years notably in the active field of health biomarker discovery. Oxylipins 18 include hundreds of structurally and stereochemically distinct lipid species which today are most 19 commonly analyzed by (ultra) high performance liquid chromatography-mass spectrometry based 20 ((U)HPLC-MS) methods. To maximize the utility of oxylipin profiling in clinical research, it is crucial to 21 understand and assess the factors contributing to the analytical and biological variability of oxylipin 22 profiles in humans. In this review, these factors and their impacts are summarized and discussed, 23 providing a framework for recommendations expected to enhance the interlaboratory comparability 24 and biological interpretation of oxylipin profiling in clinical research. 25 26 MANUSCRIPT 27 28 ACCEPTED 2 ACCEPTED MANUSCRIPT 29 1. Introduction. 30 Eicosanoids and other oxylipins represent a superfamily of signalling lipids generated from 31 arachidonic acid (AA) and related polyunsaturated fatty acids (PUFAs) through a complex network of 32 biochemical reactions involving over 50 unique and cell-specific enzymes (1). To regulate a wide array 33 of biological processes, PUFAs are converted to oxylipins via four maJor pathways (Figure 1): the 34 cyclooxygenase (COX) pathway producing prostanoids such as prostaglandins (PG) and 35 thromboxanes (Tx); the lipoxygenase (LOX) pathway producing hydroperoxy-PUFAs which are 36 rearranged into monohydroxy-PUFAs or further converted by LOX catalyzed reactions to leukotrienes 37 (LT) and numerous dihydroxy- and trihydroxy-PUFAs, including specialized pro-resolving mediators 38 (e.g. lipoxins, resolvins, protectins); the cytochrome P450 pathway (CYP), primarily producing ω-/ω-1 39 hydroxy- and epoxy-PUFAs, with the epoxides being further transformed to vicinal (i.e. adJacent or 40 1,2-)dihydroxy-PUFAs; and the nonenzymatic pathway producing various hydro(peroxy) PUFAs, 41 epoxy-PUFAs (2) as well as iso- and neuroprostanes (3, 4). In response to external stimuli 42 (e.g. bradykinin, thrombin, inflammatory insult), PUFAsMANUSCRIPT (5) and oxylipins (6) are released from 43 membrane phospholipids by phospholipases including phospholipase A2 (PLA2). Of note, hundreds 44 of structurally and strereochemically distinct oxylipins can be produced from AA and other PUFAs. 45 For instance, depending on the stimuli and the cell type, AA and EPA can be converted into PGE 2 and 46 PGE 3 respectiveley through the COX pathway or structurally distinct oxylipins (e.g. LTB 4 and LTB 5) via 47 the 5-LOX pathway (Figure 1 ). 48 The structural specificity of oxylipins leads to specificity in their biological activities, many of 49 which are still beingACCEPTED elucidated. Important to clinical research, they are notably involved in the 50 regulation of inflammation, thrombosis, endothelial function, vascular tone and insulin secretion, 51 each of these systems being either stimulated or inhibited by the different oxylipin types as 52 simplified in Figure 2 . Many oxylipins exert their biological effects by binding to cognate receptors, 53 which are members of the G protein-coupled receptor (GPCR). However, for several oxylipins such as 3 ACCEPTED MANUSCRIPT 54 epoxy-PUFAs, the receptors characterized to date cannot explain all of the biological effects elicited 55 by these compounds (7). Other known routes of oxylipin elicited effects include directly influencing 56 the open-state probability of membrane ion channels including the calcium sensitive potassium 57 channels (K Ca ) and transient receptor potential channels (TRPs) (8-10), activation of intracellular 58 transcription factors such as PPARs (11, 12), and interference with intracellular signalling pathways 59 such as NF κB (13-15). 60 Oxylipins derived from omega-3 fatty acids can be either more or less potent than or antagonistic 61 to their omega-6-derived analogs (16), e.g epoxy-omega-3-PUFA are more potent antihypertensive 62 compounds than their arachidonic acid-derived counterparts (17). Even two oxylipins derived from 63 the same PUFA can be antogonists. For instance, TxA2 and PGI 2, both derived from the oxygenation 64 of AA through the COX-dependent metabolism, respectively activate or inhibit thrombosis. Such 65 regulatory cross talk among metabolic cascades is common (18-20). Finally, depending on the 66 receptor, the tissue or the dose, a single oxylipin can also have opposite effects. For example, PGE 2 67 can exert either pro- or anti-aggregatory effects dependingMANUSCRIPT on its dose or the type of EP receptor it 68 binds to (21). Similarly, PGE 2 mediates lung inflammation in human cells (22) whereas it inhibits 69 inflammatory signalling in murine peritoneal macrophages (23) . Moreover, while PGD 2 synthesis can 70 have both pro- and anti-inflammatory impacts, and is synthesized by two convergent gene products 71 (24), regulation of this system controls the onset and resolution of inflammation in some models 72 (25). 73 While oxylipins are found in all tissues, cells are highly selective as to the type of oxylipin they 74 synthesize. For instance,ACCEPTED TxA2 is mainly produced by platelets, while the endothelium is a maJor 75 source of PGI 2. Of note, TxA synthase is also expressed in lung and macrophages and significant levels 76 of PGI synthase is found in smooth muscle cells (26, 27). PGF 2α is mainly produced by uterus, PGE 2 is 77 the maJor oxylipin generated in kidney (5) and skin (28), and the hematopeietic form of PGD 78 synthases is highly expressed in immune and inflammatory cells, but also identified in brain and 4 ACCEPTED MANUSCRIPT 79 ovary (24, 29). Some LOXs also have preferential cell distribution with LOX-5 being mainly expressed 80 in leukocytes, macrophages and dendritic cells 12/15-LOX (ALOX15 ) has a broad tissue distribution 81 (30) but is notably abundant in eosinophils and bronchial epithelium, 15-LOX2 (ALOX15B ) is highly 82 expressed in the skin and prostate, while