Annotation of Bioactive Lipid Mediators

Home , Docosapentaenoic acid, Eicosapentaenoic acid, Resolvin

Structures and Functions of Pro-Resolving Mediators

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Fig. S1. SPMs and their biosynthetic pathways.

A cluster of immunoresolvents links coagulation to innate host defense in human blood. Norris PC, Libreros S, Chiang N, Serhan CN. Sci Signal. 2017 Aug 1;10(490). 1 Specific lipid mediator signatures of human phagocytes: microparticles stimulate macrophage efferocytosis and pro-resolving mediators. Dalli J, Serhan CN. Blood. 2012 Oct 11;120(15):e60-72.

Compound Abbreviation Chemical name Bioactions Conc References

7S, 8R, 17S-trihydroxy-4Z, 9E, 11E, 13Z, 15E, Reduces vaso-obliteration and neovascularizartion 10 ng/day Connor et al. Nat. Med., 19Z-docosahexaenoic acid Protects in renal ischemic injury by limiting PMN infiltration 5 µg/mouse 2007 Duffield et al. JI, 2006 Resolvin D1 RvD1 Improves glucose tolerance and insulin sensitivity in obese-diabetic 2 µg/kg Hellmnn et al. FASEB, 2011 mice

7S, 16R, 17S-trihydroxy-4Z, 8E, 10Z, 12E, 14E, Reduces inflammatory pain 10ng/mouse Park et al. J Neurosci., Resolvin D2 RvD2 19Z-docosahexaenoic acid Enhances survival in microbial 100ng/mouse 2011 Spite et al. Nature, sepsis Protects against colitis 1µg/mouse 2009 Bento et al. JI, 2011

7S, 17S dihydroxy-4Z, 8E, 10Z, 13Z, 15E, Enhances bacterial containment synergizing with antibiotics 100ng/mouse Resolvin D5 RvD5 Chiang et al. Nature 2012 19Z-docosahexaenoic acid

10R, 17S-dihydroxy-4Z, 7Z, 11E, 13E, 15Z, Limits stroke damage and PMN entry into the brain 0.4 µg/mouse Marcheselli et al. JBC, 2003 19Z-docosahexaenoic acid Reduces vaso-obliteration and neovascularizartion Connor et al. Nat. Med., Protectin D1 PD1 10 ng/day Decreases airway eosinophil, T lymphocyte, and 2007 Levy et al. JI, 2007 proinflammatory mediators 2-200 ng/mouse

7R, 14S- Stimulates tissue regeneration in 1-100nM Maresin 1 MaR1 Serhan et al FASEB, 2012 dihydroxy-4Z,8E,10E,12Z,16Z,19Z- planaria Block inflammatory pain 10ng/ml docosahexaenoic acid 5S, 6R, 15S-trihydroxy-7E,9E,11Z,13E- Reduces inflammation; prevents connective tissue and bone loss. 6µg/tooth Serhan et al. JI, 2003 Lipoxin A4 LXA4 eicosatetraenoic acid Decrease neutrophilic inflammation, pulmonary bacterial burden 10µg/ml (drinking Karp et al. Nat. Immunol., 2004 and disease severity water)

5S, 14R, 15S-trihydroxy-6E,8Z, 10E,12E- Increases monocyte chemotaxis and reduces monocyte 0.1-100nM Maddox et al. JEM, 1996 Lipoxin B LXB 4 4 eicosatetraenoic acid adhesion Reduces PMN infiltration 26nM Takano et al. JCI, 1998

5,15-dihydroxy 5S,15S-dihydroxy-6E,8Z,11Z,13E-eicosatetraenoic 5,15-diHETE eicosatetraenoic acid acid

5S, 12R, 18R-trihydroxy-6Z, 8E, 10E, 14Z, Reduces vaso-obliteration and 10 ng/day Connor et al. Nat. Med., Resolvin E1 RvE1 16E-eicosapentaenoic acid neovascularizartion Reduces PMN infiltration 20ng/mouse 2007 Oh et al. JCI, 2010 Reduces persistent inflammatory pain 1ng/mouse Xu et al. Nat. Med., 2010

5S, 18R-trihydroxy-6E, 8Z, 11Z, 14Z, Reduces PMN infiltration 10 ng/mouse Tjonahen et al. Chem Biol, 2006 Resolvin E2 RvE2 16E-eicosapentaenoic acid

5S, 6R, 15S-trihydroxy-7E,9E,11Z,13E, Lipoxin A LXA 5 5 17Z-eicosapentaenoic acid

5S, 14R, 15S-trihydroxy-6E,8Z,10E, Lipoxin B LXB 5 5 12E,17Z-eicosapentaenoic acid

5S,12R-dihydroxy-6Z, 8E,10E, 14Z-eicosatetraenoic Induces vascular leakage 10-20n Thureson-Klein et al. Tiss. Cell, Leukotriene B LTB 4 4 acid Induces leukocyte adhesion M 4nM 1986 Dahlen et al. PNAS, 1981

5S,12R-dihydroxy-6Z, 8E,10E,14Z, 17Z- Induces endothelial cell permeability 0.1-1µM Terano et al. Prostaglandins, 1984 Leukotriene B LTB 5 5 eicosapentaenoic acid Stimulates PMN chemotaxis 0.1-1µM Moreno J Pharmacol Exp Ther., 2009 9S,15S-dihydroxy-11-oxo-5Z,13E-prostadienoic Induces vaso-relaxation Induces 0.1-1µM Braun et al. Cir Res, 1992 Prostaglandin D PGD 2 2 acid eosinophil migration 0.1-1µM Schratl et al. JI, 2007

Prostaglandin E2 PGE2 9-oxo-11R,15S-dihydroxy-5Z,13E-prostadienoic Induces vascular leakage 1µg Chan et al. J Inv Derma, 1985 acid 9S,11R,15S-trihydroxy-5Z,13E-prostadienoic Induces vasoconstriction 10µg/Kg Yu et al. PNAS, 2009 Prostaglandin F PGF 2a 2a acid Uterine contraction 1µM Hsia et al. Endocrinology, 2011

9S,11,15S-trihydroxy-thromboxa-5Z,13E-dien-1-oic Induces PMN chemotaxis 0.5µg/ml Kitchen et al Prostaglandins, Thromboxane B TXB 2 2 acid Potentiated contractile response in mesenteric arteries 1µM 1978 Okamura et al. BJP, 1988

9S,15S-dihydroxy-11-oxo-5Z,13E17Z-prostatrienoic Induces peripheral vasodilatation Modulatiuon of 0.24-1µg/Kg/min Wendling et al. Prostaglandins, 1981 Prostaglandin D PGD 3 3 acid autonomic nerve transmission 1-10µg/Kg Hemker et al. Eu J. Pharma., 1980

9-oxo-11R,15S-dihydroxy-5Z,13E,17Z-prostatrienoic Induces COX-2 and IL6 expression in macrophages Increases 0.03-3µM Bagga et al. PNAS, 2003 Shimizu Prostaglandin E PGE 3 s acid plasma adrenaline when administered in the brain 300nM et al. Eu J. Pharma., 2009

Prostaglandin F3a PGF3a 9S,11R,15S-trihydroxy-5Z,13E-prostatrienoic acid Protexts from gastric mucosal injury 16µM/Kg Faust et al. Prostaglandins, 1989

Induces COX-2 and IL6 expression in macrophages 0.03-3µM Bagga et al. PNAS, 2003 Thromboxane B TXB 9S,11,15S-trihydroxy-thromboxa-5Z,13E,17Z-dien-1-oic 3 3 acid

11 Table S1 References

Bagga D, Wang L, Farias-Eisner R, Glaspy JA, Reddy ST. Differential effects of prostaglandin derived from omega-6 and omega-3 polyunsaturated fatty acids on COX-2 expression and IL-6 secretion. Proc Natl Acad Sci U S A. 2003;100(4):1751-1756. Bento AF, Claudino RF, Dutra RC, Marcon R, Calixto JB. Omega-3 fatty acid-derived mediators 17(R)-hydroxy docosahexaenoic acid, aspirin-triggered resolvin D1 and resolvin D2 prevent experimental colitis in mice. J Immunol. 2011;187:1957-1969. Braun M, Schror K. Prostaglandin D2 relaxes bovine coronary arteries by endothelium- dependent nitric oxide-mediated cGMP formation. Circ Res. 1992;71(6):1305-1313. Chan CC, Ford-Hutchinson A. Effects of synthetic leukotrienes on local blood flow and vascular permeability in porcine skin. J Invest Dermatol. 1985;84(2):154-157. Chiang N, Fredman G, Bäckhed F, et al. Infection regulates pro-resolving mediators that lower antibiotic requirements. Nature. 2012;484:524-528. Connor KM, SanGiovanni JP, Lofqvist C, et al. Increased dietary intake of omega-3- polyunsaturated fatty acids reduces pathological retinal angiogenesis. Nat Med. 2007;13:868- 873. Dahlen SE, Bjork J, Hedqvist P, et al. Leukotrienes promote plasma leakage and leukocyte adhesion in postcapillary venules: in vivo effects with relevance to the acute inflammatory response. Proc Natl Acad Sci U S A. 1981;78(6):3887-3891. Duffield JS, Hong S, Vaidya V, et al. Resolvin D series and protectin D1 mitigate acute kidney injury. J Immunol. 2006;177:5902-5911. Faust TW, Lee E, Redfern JS, Feldman M. Effect of prostaglandin F3 alpha on gastric mucosal injury by ethanol in rats: comparison with prostaglandin F2 alpha. Prostaglandins. 1989;37(4):493-504. Hellmann J, Tang Y, Kosuri M, Bhatnagar A, Spite M. Resolvin D1 decreases adipose tissue macrophage accumulation and imiproves insulin sensitivity in obese-diabetic mice. FASEB J. 2011;25:2399-2407. Hemker DP, Aiken JW. Effects of prostaglandin D3 on nerve transmission in nictitating membrane of cats. Eur J Pharmacol. 1980;67(1):155-158. Hsia SM, Wang KL, Wang PS. Effects of resveratrol, a grape polyphenol, on uterine contraction and Ca(2)+ mobilization in rats in vivo and in vitro. Endocrinology. 2011;152(5):2090-2099. Karp CL, Flick LM, Park KW, et al. Defective lipoxin-mediated anti-inflammatory activity in the cystic fibrosis airway. Nat Immunol. 2004;5:388-392. Kitchen EA, Boot JR, Dawson W. Chemotactic activity of thromboxane B2, prostaglandins and their metabolites for polymorphonuclear leucocytes. Prostaglandins. 1978;16(2):239-244. Levy BD, Kohli P, Gotlinger K, et al. Protectin D1 is generated in asthma and dampens airway inflammation and hyper-responsiveness. J Immunol. 2007;178:496-502.

3 Maddox JF, Serhan CN. Lipoxin A4 and B4 are potent stimuli for human monocyte migration and adhesion: selective inactivation by dehydrogenation and reduction. J Exp Med. 1996;183:137- 146. Marcheselli VL, Hong S, Lukiw WJ, et al. Novel docosanoids inhibit brain ischemia- reperfusion-mediated leukocyte infiltration and pro-inflammatory gene expression. J Biol Chem. 2003;278:43807-43817. Moreno JJ. Differential effects of arachidonic and eicosapentaenoic acid-derived eicosanoids on polymorphonuclear transmigration across endothelial cell cultures. J Pharmacol Exp Ther. 2009;331:1111-1117. Oh SF, Pillai PS, Recchiuti A, Yang R, Serhan CN. Pro-resolving actions and stereoselective biosynthesis of 18S E-series resolvins in human leukocytes and murine inflammation. J Clin Invest. 2011;121:569-581. Okamura T, Nakajima M, Toda N. Neuroeffector actions of thromboxane B2 in dog isolated mesenteric arteries. Br J Pharmacol. 1988;93(2):367-374. Park CK, Lü N, Xu ZZ, Liu T, Serhan CN, Ji RR. Resolving TRPV1 and TNF-α-mediated spinal cord synaptic plasticity and inflammatory pain with neuroprotectin D1. J Neurosci. 2011;31:15072-15085. Schratl P, Royer JF, Kostenis E, et al. The role of the prostaglandin D2 receptor, DP, in eosinophil trafficking. J Immunol. 2007;179:4792-4799. Serhan CN, Dalli J, Karamnov S, et al. Macrophage pro-resolving mediator maresin 1 stimulates tissue regeneration and controls pain. FASEB J. 2012;26:1755-1765. Serhan CN, Jain A, Marleau S, et al. Reduced inflammation and tissue damage in transgenic rabbits overexpressing 15-lipoxygenase and endogenous anti-inflammatory lipid mediators. J Immunol. 2003;171:6856-6865. Shimizu T, Yokotani K. Effects of centrally administered prostaglandin E(3) and thromboxane A(3) on plasma noradrenaline and adrenaline in rats: comparison with prostaglandin E(2) and thromboxane A(2). Eur J Pharmacol. 2009;611(1-3):30-34. Spite M, Norling LV, Summers L, et al. Resolvin D2 is a potent regulator of leukocytes and controls microbial sepsis. Nature. 2009;461:1287-1291. Takano T, Clish CB, Gronert K, Petasis N, Serhan CN. Neutrophil-mediated changes in vascular permeability are inhibited by topical application of aspirin-triggered 15-epi-lipoxin A4 and novel lipoxin B4 stable analogues. J Clin Invest. 1998;101:819-826. Terano T, Salmon JA, Moncada S. Biosynthesis and biological activity of leukotriene B5. Prostaglandins. 1984;27(2):217-232. Thureson-Klein A, Hedqvist P, Lindbom L. Leukocyte diapedesis and plasma extravasation after leukotriene B4: lack of structural injury to the endothelium. Tissue Cell. 1986;18(1):1-12. Tjonahen E, Oh SF, Siegelman J, et al. Resolvin E2: Identification and anti-inflammatory actions: pivotal role of human 5-lipoxygenase in resolvin E series biosynthesis. Chem Biol. 2006;13:1193-1202.

4 Wendling MG, DuCharme DW. Cardiovascular effects of prostaglandin D3 and D2 in anesthetized dogs. Prostaglandins. 1981;22(2):235-243. Xu Z-Z, Zhang L, Liu T, et al. Resolvins RvE1 and RvD1 attenuate inflammatory pain via central and peripheral actions. Nat Med. 2010;16:592-597. Yu Y, Lucitt MB, Stubbe J, et al. Prostaglandin F2alpha elevates blood pressure and promotes atherosclerosis. Proc Natl Acad Sci U S A. 2009;106(19):7985-7990.

5 DHA bioactive metabolome (updated 2019): SPM (See reference 1)

Full name Abbreviation Chemical name Pro-inflammatory or Pro-resolving actions Organ protection Pain pro-resolving Resolvin D1 RvD1 7S,8R,17S-trihydroxy- pro-resolving Stops PMN migration Peritonitis, air pouch, CLP/sepsis, bacterial Analgesic 4Z,9E,11E,13Z,15E,19Z- Reduces pro-inflammatory cytokines infections, I/R injury, burn, diabetes, obesity, docosahexaenoic acid Enhances MF phagocytosis & efferocytosis colitis, kidney injury, airway, vascular, ocular & Inhibits TRP channels neuroinflammation, Alzheimer’s disease, Modulates T cell responses arthritis, atherosclerosis, tumor growth, Reduces IgE production vagotomy, fibromyalgia, Sjögren's syndrome, myocardial infarct Resolvin D2 RvD2 7S,16R,17S-trihydroxy- pro-resolving Stops PMN migration Peritonitis, air pouch, CLP/sepsis, bacterial Analgesic 4Z,8E,10Z,12E,14E,19Z- Reduces pro-inflammatory cytokines infections, I/R injury, burn, atherosclerosis, docosahexaenoic acid Enhances MF phagocytosis & efferocytosis periodontitis, obesity, colitis, fibromyalgia, Inhibits TRP channels reduces tumor growth. Suppresses NLRP3 inflammasome Modulates T cell responses Resolvin D3 RvD3 4S,11R,17S-trihydroxy- pro-resolving Blocks PMN transmigration Peritonitis, arthritis, bacterial infections 5Z,7E,9E,13Z,15E,19Z- Enhances MF phagocytosis & efferocytosis docosahexaenoic acid Restores epithelial barrier & function Resolvin D4 RvD4 4S,5R,17S-trihydroxy- pro-resolving Enhances fibroblast phagocytosis Peritonitis, air pouch. I/R injury 6E,8E,10Z,13Z,15E,19Z- docosahexaenoic acid Resolvin D5 RvD5 7S,17S-dihydroxy- pro-resolving Enhances MF and PMN phagocytosis Bacterial infections 4Z,8E,10Z,13Z,15Z,19E- docosahexaenoic acid Resolvin D6 RvD6 4S,17S-dihydroxy- 5E,7E,10Z,13Z,15E,19Z- docosahexaenoic acid Protectin D1 PD1 10R,17S-dihydroxy- pro-resolving Limits PMN migration Peritonitis, cerebral I/R injury, Analgesic 4Z,7Z,11E,13E,15Z,19Z- Reduces pro-inflammatory cytokines neuroinflammation, Alzheimer’s disease, docosahexaenoic acid Enhances MF phagocytosis & efferocytosis influenza Maresin 1 MaR1 7R,14S-dihydroxy- pro-resolving Reduces pro-inflammatory cytokines Peritonitis, Planaria head regeneration, spinal Analgesic 4Z,8E,10E,12Z,16Z,19Z- Enhances MF phagocytosis and efferocytosis cord injury, fibrosis, skin inflammation, docosahexaenoic acid Inhibits TRP channels atherosclerosis, obesity, CLP/sepsis, liver Enhances platelet aggregation injury, airway inflammation Regulates autophagy Modulates T cell responses Maresin 2 MaR1 13R,14S-dihydroxy- pro-resolving Enhances MF phagocytosis & efferocytosis Peritonitis 4Z,7Z,9E,11E,16Z,19Z- docosahexaenoic acid 17S-HDHA 17S-hydroxy- Pro-resolving Enhances antibody-mediated immune response Reduced adipose tissue expression of Analgesic (6) 4Z,7Z,10Z,13Z,15E,19Z- against influenza virus (2) inflammatory cytokines, increased adiponectin docosahexaenoic acid expression, and improved glucose tolerance parallel to insulin sensitivity in obese mice (3) Alleviate experimental colitis (4) and colitis (5)

6 DHA bioactive metabolome (updated 2019): Aspirin-triggered SPM (See reference 1)

Full name Abbreviation Chemical name Pro-inflammatory or Pro-resolving actions Organ protection Pain pro-resolving

Aspirin- AT-RvD1 7S,8R,17R-trihydroxy- Pro-resolving Stops PMN migration Peritonitis, air pouch, colitis, arthritis, Analgesic triggered- 4Z,9E,11E,13Z,15E,19Z- Reduces pro-inflammatory cytokines temporomandibular joint inflammation, Resolvin D1 docosahexaenoic acid Enhances MF phagocytosis & acute lung injury & immune complex- efferocytosis induced lung injury, asthma, surgery- Modulates T cell responses induced cognitive decline, acute kidney Accelerates keratinocyte healing injury, kidney I/R injury, fibromyalgia, Improves epithelial and endothelial Sjögren's syndrome. Influenza, barrier integrity pneumonia, stromal keratitis, Chagas Regulates conjunctival goblet cell disease secretion Attenuates vascular smooth muscle cell migration

Aspirin- AT-RvD2 7S,16R,17R-trihydroxy- Pro-resolving Reduces microglial cell cytokine Peritonitis, air pouch triggered- 4Z,8E,10Z,12E,14E,19Z- expression Resolvin D2 docosahexaenoic acid

Aspirin- AT-RvD3 4S,11R,17R-trihydroxy- Pro-resolving Blocks PMN transmigration Peritonitis, air pouch, bacterial infections triggered- 5Z,7E,9E,13Z,15E,19Z- Enhances MF phagocytosis & Resolvin D3 docosahexaenoic acid efferocytosis Restores epithelial barrier & function

Aspirin- AT-RvD4 4S,5R,17R-trihydroxy- Pro-resolving Reduces microglial cell cytokine Peritonitis, air pouch triggered- 6E,8E,10Z,13Z,15E,19Z- expression Resolvin D4 docosahexaenoic acid

Aspirin- AT-RvD5 7S,17R-dihydroxy- Pro-resolving Reduces microglial cell cytokine Peritonitis, air pouch triggered- 4Z,8E,10Z,13Z,15Z,19E- expression Resolvin D5 docosahexaenoic acid

Aspirin- AT-RvD6 4S,17R-dihydroxy- Pro-resolving Reduces microglial cell cytokine Peritonitis, air pouch triggered- 5E,7E,10Z,13Z,15E,19Z- expression Resolvin D6 docosahexaenoic acid

Aspirin- AT-PD1 10R,17R-dihydroxy- Pro-resolving Enhances MF phagocytosis & Peritonitis, cerebral I/R injury triggered- 4Z,7Z,11E,13E,15Z,19Z- efferocytosis Protectin D1 docosahexaenoic acid Limits transendothelial PMN migration

7 DHA bioactive metabolome (updated 2019): Cysteinyl conjugated SPM (See reference 7,8)

Full name Abbreviation Chemical name Pro-inflammatory or Pro-resolving actions Organ protection pro-resolving Maresin conjugates in MCTR1 13R-glutathionyl-14S-hydroxy- pro-resolving Enhances MF phagocytosis & Accelerates planarian tissue regeneration 1 4Z,7Z,9E,11E,16Z,19Z- efferocytosis regeneration, E. coli peritonitis docosahexaenoic acid

Maresin conjugates in MCTR2 13R-cysteinylglycinyl-14S-hydroxy- pro-resolving Enhances MF phagocytosis & Accelerates planarian tissue regeneration 2 4Z,7Z,9E,11E,16Z,19Z- efferocytosis regeneration, E. coli peritonitis docosahexaenoic acid

Maresin conjugates in MCTR3 13R-cysteinyl-14S-hydroxy- pro-resolving Enhances MF phagocytosis & Accelerates planarian tissue regeneration 3 4Z,7Z,9E,11E,16Z,19Z- efferocytosis regeneration, E. coli peritonitis docosahexaenoic acid

Resolvin conjugates in RCTR1 8R-glutathionyl-7S,17S-dihydroxy- pro-resolving Stop PMN chemotaxis Accelerates planarian tissue regeneration 1 4Z,9E,11E,13Z,15E,19Z- Enhances MF phagocytosis & regeneration, E. coli peritonis, docosahexaenoic acid efferocytosis, Regulates cytokines I/R injury

Resolvin conjugates in RCTR2 8R-cysteinylglycinyl-7S,17S-dihydroxy- pro-resolving Stop PMN chemotaxis Accelerates planarian tissue regeneration 2 4Z,9E,11E,13Z,15E,19Z- Enhances MF phagocytosis & regeneration, E. coli peritonis, docosahexaenoic acid efferocytosis, Regulates cytokines I/R injury

Resolvin conjugates in RCTR3 8R-cysteinyl-7S,17S-dihydroxy- pro-resolving Stop PMN chemotaxis Accelerates planarian tissue regeneration 3 4Z,9E,11E,13Z,15E,19Z- Enhances MF phagocytosis & regeneration, E. coli peritonis, docosahexaenoic acid efferocytosis, Regulates cytokines I/R injury

Protectin conjugates in PCTR1 16R-glutathionyl-17S-hydroxy- pro-resolving Promotes human monocyte and Accelerates planarian regeneration tissue regeneration 1 4Z,7Z,10Z,12E,14E,19Z- macrophage migration E. coli peritonis and vagotomy docosahexaenoic acid Enhances MF phagocytosis & efferocytosis, Increases M2 MF

Protectin conjugates in PCTR2 16R-cysteinylglycinyl-17S-hydroxy- pro-resolving Enhances MF phagocytosis & Accelerates planarian regeneration tissue regeneration 2 4Z,7Z,10Z,12E,14E,19Z- efferocytosis, docosahexaenoic acid

Protectin conjugates in PCTR3 16R-cysteinyl-17S-hydroxy- pro-resolving Accelerates planarian regeneration tissue regeneration 3 4Z,7Z,10Z,12E,14E,19Z- docosahexaenoic acid

8 EPA bioactive metabolome (updated 2019) (See reference 9.10)

Full name Abbreviation Chemical name Pro-inflammatory or Pro-resolving actions Organ protection Pain pro-resolving

Resolvin E1 RvE1 5S,12R,18R-trihydroxy- pro-resolving Stops PMN migration Peritonitis, air pouch, CLP/sepsis, bacterial Analgesic 6Z,8E,10E,14Z,16E- Reduces pro-inflammatory & fungal infections, I/R injury, burn, diabetes, eicosapentaenoic acid cytokines obesity, colitis, lung inflammation, kidney Enhances MF phagocytosis & injury, vascular inflammation, efferocytosis neuroinflammation, Alzheimer’s disease, Inhibits TRP channels arthritis, atherosclerosis, reduces tumor Modulates T cell responses growth

18S-resolvin E1 18S-RvE1 5S,12R,18S-trihydroxy- pro-resolving Stops PMN migration Peritonitis 6Z,8E,10E,14Z,16E- Reduces pro-inflammatory eicosapentaenoic acid cytokines Enhances MF phagocytosis & efferocytosis

Resolvin E2 RvE2 5S,18R-dihydroxy- pro-resolving Stops PMN migration Peritonitis 6E,8Z,11Z,14Z,16E- Down-regulates leukocyte eicosapentaenoic acid; integrins

18R-Resolvin E3 18R-RvE3 17R,18R-dihydroxy- pro-resolving Stops PMN migration Peritonitis 5Z,8Z,11Z,13E,15E- eicosapentaenoic acid

18S-Resolvin E3 18S-RvE3 17R,18S-dihydroxy- Stops PMN migration Peritonitis 5Z,8Z,11Z,13E,15E- eicosapentaenoic acid

18R-HEPE 18R-hydroxy- pro-resolving Prevents pressure overload-induced 5Z,8Z,11Z,14Z,16E- maladaptive cardiac remodeling (11) eicosapentaenoic acid Protects against CXCR4-associated melanoma metastasis (12) Restore mitochondrial function in inflammation (13) Decreases monocyte adhesion to endothelial cells (14)

9 AA bioactive metabolome: Lipoxins (See reference 15, 16)

Full name Abbreviation Chemical name Pro-inflammatory or Actions Pain pro-resolving

AA metabolome

Lipoxin A4 LXA4 5S,6R,15S-trihydroxy-7E,9E,11Z,13E- pro-resolving Stops PMN migration, enhances MF Analgesic eicosatetraenoic acid phagocytosis & efferocytosis

Lipoxin B4 LXB4 5S,14R,15S-trihydroxy- pro-resolving Regulates antibody production, regulates NLRP3 Analgesic 6E,8Z,10E,12E-eicosatetraenoic acid inflammasome, T cell responses

Aspirin-triggered AT-LXA4 5S,6R,15R-trihydroxy-7E,9E,11Z,13E- pro-resolving Stops PMN migration, enhances phagocytosis Analgesic lipoxin A4 eicosatetraenoic acid

Aspirin-triggered AT-LXB4 5S,14R,15R-trihydroxy- pro-resolving lipoxin B4 6E,8Z,10E,12E-eicosatetraenoic acid

10 References

(1) Protectins and maresins: New pro-resolving families of mediators in acute inflammation and resolution bioactive metabolome. Serhan CN, Dalli J, Colas RA, Winkler JW, Chiang N. Biochim Biophys Acta. 2015 Apr;1851(4):397-413 (2) The specialized proresolving mediator 17-HDHA enhances the antibody-mediated immune response against influenza virus: a new class of adjuvant? Ramon S, Baker SF, Sahler JM, Kim N, Feldsott EA, Serhan CN, Martínez-Sobrido L, Topham DJ, Phipps RP. J Immunol. 2014 Dec 15;193(12):6031-40. (3) mpaired local production of proresolving lipid mediators in obesity and 17-HDHA as a potential treatment for obesity-associated inflammation. Neuhofer A, Zeyda M, Mascher D, Itariu BK, Murano I, Leitner L, Hochbrugger EE, Fraisl P, Cinti S, Serhan CN, Stulnig TM. Diabetes. 2013 Jun;62(6):1945-56. (4) Omega-6 docosapentaenoic acid-derived resolvins and 17-hydroxydocosahexaenoic acid modulate macrophage function and alleviate experimental colitis. Chiu CY, Gomolka B, Dierkes C, Huang NR, Schroeder M, Purschke M, Manstein D, Dangi B, Weylandt KH. Inflamm Res. 2012 Sep;61(9):967-76. (5) The precursor of resolvin D series and aspirin-triggered resolvin D1 display anti-hyperalgesic properties in adjuvant-induced arthritis in rats. Lima-Garcia JF, Dutra RC, da Silva K, Motta EM, Campos MM, Calixto JB. Br J Pharmacol. 2011 (6) Association of the resolvin precursor 17-HDHA, but not D- or E- series resolvins, with heat pain sensitivity and osteoarthritis pain in humans. Valdes AM, Ravipati S, Menni C, Abhishek A, Metrustry S, Harris J, Nessa A, Williams FMK, Spector TD, Doherty M, Chapman V, Barrett DA. Sci Rep. 2017 Sep 7;7(1):10748. (7) New pro-resolving n-3 mediators bridge resolution of infectious inflammation to tissue regeneration. Serhan CN, Chiang N, Dalli J. Mol Aspects Med. 2018 Dec;64:1-17. (8) Identification and Complete Stereochemical Assignments of the New Resolvin Conjugates in Tissue Regeneration in Human Tissues that Stimulate Proresolving Phagocyte Functions and Tissue Regeneration. de la Rosa X, Norris PC, Chiang N, Rodriguez AR, Spur BW, Serhan CN. Am J Pathol. 2018 Apr;188(4):950-966. (9) Resolving inflammation: dual anti-inflammatory and pro-resolution lipid mediators. Serhan CN, Chiang N, Van Dyke TE. Nat Rev Immunol. 2008 May;8(5):349- 61. (10) Omega-3 fatty acid-derived mediators that control inflammation and tissue homeostasis. Ishihara T, Yoshida M, Arita M. Int Immunol. 2019 Feb 17. (11) 18-HEPE, an n-3 fatty acid metabolite released by macrophages, prevents pressure overload-induced maladaptive cardiac remodeling. Endo J, Sano M, Isobe Y, Fukuda K, Kang JX, Arai H, Arita M. J Exp Med. 2014 Jul 28;211(8):1673-87. (12) An omega-3 polyunsaturated fatty acid derivative, 18-HEPE, protects against CXCR4-associated melanoma metastasis. Li J, Chen CY, Arita M, Kim K, Li X, Zhang H, Kang JX. Carcinogenesis. 2018 Dec 13;39(11):1380-1388. (13) Resolvin E1 and its precursor 18R-HEPE restore mitochondrial function in inflammation. Hecker M, Sommer N, Foch S, Hecker A, Hackstein H, Witzenrath M, Weissmann N, Seeger W, Mayer K. Biochim Biophys Acta Mol Cell Biol Lipids. 2018 Sep;1863(9):1016-1028. (14) Metabolic profiling of murine plasma reveals eicosapentaenoic acid metabolites protecting against endothelial activation and atherosclerosis. Liu Y, Fang X, Zhang X, Huang J, He J, Peng L, Ye C, Wang Y, Xue F, Ai D, Li D, Zhu Y. Br J Pharmacol. 2018 Apr;175(8):1190-1204. (15) Leukotrienes and lipoxins: structures, biosynthesis, and biological effects. Samuelsson B, Dahlén SE, Lindgren JA, Rouzer CA, Serhan CN. Science. 1987 Sep 4;237(4819):1171-6. (16) Lipoxins and novel aspirin-triggered 15-epi-lipoxins (ATL): a jungle of cell-cell interactions or a therapeutic opportunity? Serhan CN. Prostaglandins. 1997 Feb;53(2):107-37.

11 402 C.N. Serhan et al. / Biochimica et Biophysica Acta 1851 (2015) 397–413

Table 1 Production and actions of Protectins and Maresins.

Mediator Species/system Concentration/amount Action(s) Function In vivo production investigated

(A) Protectins NPD1/PD1 Mouse/peritonitis 300 ng/mouse Inhibits neutrophil recruitment; regulates chemokine/cytokine production [43,50] 300 ng/mouse Promotes lymphatic removal of phagocytes [76] 100 ng/mouse Regulates T-cell migration [48] Mouse/asthma 2–200 ng/mouse Protects from lung damage, airway inflammation and hyperresponsiveness [137] 100 ng/mouse Accelerates catabasis of Th2-mediated inflammation [138] 11 pg/mouse Toll-like receptor 7 triggers production of PD1 [138] Human/asthma ~2 ng/mL of exhaled PD1 is reduced in human breath condensate asthma [137] from healthy subjects Human 10 ng/106 cells in Dysregulation of PD1 in eosinophils/ healthy subjects eosinophils from asthma asthma patients [139] Mouse/ 5 μg/mouse Rescues delayed resolution in eosinophil eosinophils depleted mice [140] Mouse/kidney 3.5–35 μg/mouse Protects from ischemia–reperfusion-induced ischemia– kidney damage and loss of function; regulates reperfusion macrophages [73,141] 160 pg/kidney Endogenous formation in kidney increased for ω − 3 PUFA [73] Mouse/ 10 ng/mouse Protects against neovascularization [71] retinopathy Ischemic stroke 100 ng/mouse Inhibits leukocyte infiltration, NFκB and cyclooxygenase-2 induction [14,44] Human/ 5 pmol/mg protein Diminished PD1 Alzheimer's production in human disease Alzheimer's disease [142] Mouse/liver injury 100 nM Protects from necro-inflammatory liver injury [143] Rabbit/corneal 100 ng/rabbit Promotes regeneration of corneal nerves injury following surgery [144] Mouse/Stem cells 50 nM Protects stem cells from oxidative stress [75] Mouse/Lyme 0.1 pg/mg of tissue PD1 produced during the disease resolution of Lyme disease [145] Rat/optic nerve 5 μg/rat Inhibits cell ganglion death [146] injury Mouse/obesity 100 nM Regulates adiponectin in adipose tissue of obese (ob/ob) mice [147] ~150 pg/mouse Generated in adipose tissue [147] Mouse/neuronal 1 ng/mL Reduces TRPV1-mediated inflammatory pain plasticity [70] Mouse/neuronal 300 ng/mouse Antinociceptive without associated tolerance plasticity [148] Mouse/herpes 300 ng/mouse Reduction of stromal keratitis and simplex virus 1 neovascularization [149] Rats & mouse/ 50 nM Attenuates evoked hippocampal seizures [150] epilepsy Fat-1 mouse/ 0.2 ng/mg colon Physiologically active colitis tissue levels produced in colon of transgenic mice [151] Human/ARPE-19 50 nM Anti-apoptotic for ARPE-19 cell survival [152] cells Human MΦ and 0.1–10 nM Promotes bacterial PMN/phagocytosis clearance [57] Mouse/E. coli 50 ng/mouse Protects from hypothermia [57] and reduces peritonitis bacterial titers 1 ng/mouse Lower levels in higher bacterial challenge delayed resolution [57] 10S, 17S diHDHA (PDX) Mouse/peritonitis 1–100 ng/mouse Inhibits neutrophil recruitment; regulates chemokine/ cytokine production [50] Mouse/H1N1 1.0 μg/mouse Antiviral [153] 2.7 μM Attenuates translocation [153] Human/platelets 0.3–1.0 μM Inhibits platelet aggregation [64,136] Mouse/acute lung 200 μg/mouse Enhances neutrophil apoptosis [154] injury

12 C.N. Serhan et al. / Biochimica et Biophysica Acta 1851 (2015) 397–413 403

Table 1 (continued)

Mediator Species/system Concentration/amount Action(s) Function In vivo production investigated

PDX Mouse/diabetes 1 μg/mouse Stimulates skeletal muscle IL-6 release; improves insulin sensitivity [67] AT-PD1 Mouse/peritonitis 0.01–100 ng/mouse Regulates PMN inﬁltration [49] Human MΦ 0.01–100 nM Promotes efferocytosis [49] Rat/stroke 333 μg/kg Reduces brain infarction and edema in stroke [155]

(B) Maresins MaR1 Mouse/peritonitis 0.1 ng/mouse Inhibits neutrophil recruitment; enhances phagocyte clearance [55] Planaria/ 100 nM Stimulates tissue regeneration regeneration in planaria [54] Mouse/pain 10 ng/mouse Antinociceptive [54] human/rheuma- 7 pg/50 μLSF Identified in synovial fluid toid arthritis extracts [113] Mouse/peritonitis 26 pg/mouse Elevated levels with CO inhalation [134] Human MΦ 10 nM Phenotype switch [115] 24 pg/2 × 105 MΦ Macrophage phagocytosis of apoptotic PMN [112] Mouse/colitis 300 ng/mouse Reduced PMN infiltration and cytokine expression as well as NF-κB activation [156] human MΦ 0.1–100 nM Promotes efferocytosis [114,157] human bronchial 100–200 nM Reduces cytokine release with organic dust epithelial cells [158]

Protectins and maresins: New pro-resolving families of mediators in acute inflammation and resolution bioactive metabolome. Serhan CN, Dalli J, Colas RA, Winkler JW, Chiang N. Biochim Biophys Acta. 2015 Apr;1851(4):397-413

13 236 LIBREROS

TABLE 1 Specialized pro-resolving lipid mediator families biosynthesized from their parent polyunsaturated fatty acids

Full name Abbreviation Chemical name Structure Pro-resolving actions DHA derived SPMs Protectin D1 PD1 10R,17S-dihydroxy- Reduces pro-inflammatory cytokines 4Z,7Z,11E,13E,15Z,19Z- Limits PMN migration docosahexaenoic acid OH COOH Enhances MΦ phagocytosis & efferocytosis

10S,17S- PDX 10S,17(S)-dihydroxy- Limits PMN infiltration diHDHA 4Z,7Z,11E,13Z,15E,19Z- Reduces platelets aggregation docosahexaenoic OH acid COOH

Maresin 1 MaR1 7R,14S-dihydroxy- Reduces pro-inflammatory cytokines OH 4Z,8E,10E,12Z,16Z,19Z- OH Enhances MΦ phagocytosis and efferocytosis docosahexaenoic acid Inhibits TRP channels Enhances platelet aggregation Regulates autophagy COOH Modulates T cell responses Maresin 2 MaR2 13R,14S-dihydroxy- Enhances MΦ phagocytosis & efferocytosis 4Z,7Z,9E,11E,16Z,19Z- HO

docosahexaenoic acid OH COOH

Resolvin D1 RvD1 7S,8R,17S-trihydroxy- HO Stops PMN migration 4Z,9E,11E,13Z,15E,19Z- Reduces pro-inflammatory cytokines COOH docosahexaenoic OH Enhances MΦ phagocytosis & efferocytosis acid Inhibits TRP channels

HO Modulates T cell responses Reduces IgE production

Resolvin D2 RvD2 7S,16R,17S-trihydroxy- COOH Stops PMN migration 4Z,8E,10Z,12E,14E,19Z- OH Reduces pro-inflammatory cytokines docosahexaenoic Enhances MΦ phagocytosis & efferocytosis acid OH Inhibits TRP channels

HO Suppresses NLRP3 inflammasome Modulates T cell responses

Resolvin D3 RvD3 4S,11R,17S-trihydroxy- OH Blocks PMN transmigration 5Z,7E,9E,13Z,15E,19Z- HO COOH Enhances MΦ phagocytosis & efferocytosis docosahexaenoic Restores epithelial barrier & function acid HO

Resolvin D4 RvD4 4S,5R,17S-trihydroxy- HO COOH Enhances fibroblast phagocytosis 6E,8E,10Z,13Z,15E,19Z- Enhances E. coli phagocytosis by neutrophils and OH docosahexaenoic monocytes acid HO

Resolvin D5 RvD5 7S,17S-dihydroxy- COOH Enhances MΦ and PMN phagocytosis 4Z,8E,10Z,13Z,15Z,19E- HO docosahexaenoic HO acid

Resolvin D6 RvD6 4S,17S-dihydroxy- HO COOH 5E,7E,10Z,13Z,15E,19Z- docosahexaenoic acid OH

EPA derived SPMs

Resolvin E1 RvE1 5S,12R,18R-trihydroxy- OH Stops PMN migration 6Z,8E,10E,14Z,16E- Reduces pro-inflammatory cytokines HO eicosapentaenoic OH Enhances MΦ phagocytosis & efferocytosis acid COOH Inhibits TRP channels Modulates T cell responses (Continues)

14 LIBREROS 237

TABLE 1 (Continued)

Full name Abbreviation Chemical name Structure Pro-resolving actions

Resolvin E2 RvE2 5S,18R-dihydroxy- OH Stops PMN migration 6E,8Z,11Z,14Z,16E- Down-regulates leukocyte integrins eicosapentaenoic COOH acid; HO

AA derived SPMs Φ Lipoxin A4 LXA4 5S,6R,15S-trihydroxy- HO OH Stops PMN migration, enhances M 7E,9E,11Z,13E- COOH phagocytosis & efferocytosis eicosatetraenoic acid OH

Lipoxin B4 LXB4 5S,14R,15S-trihydroxy- OH Regulates Ab production, regulates NLRP3 6E,8Z,10E,12E- COOH inflammasome, T cell responses eicosatetraenoic acid HO OH

(S) Pathway COOH Markers AB14S-HDHA RvD1 17-HDHA LXA4 14-HDHA PDX OOH 12-LOX RvE1 DHA (S) COOH O 2 ALX/FPR2 14S-HpDHA O2 Resolvins ERV1 Obesity 15-LOX (D Series) (S) OOH 5-LOX Autommunity

COOH Maresin 1 (MaR1) Viral 17S-HpDHA B Cell Infections Enzymatic Reduction Epoxidation GPCR37 (S) (S) OH (S) O Markers PD1 COOH Pro-inflammatory COOH cytokines 16S, 17S-epoxy-protectin 17S-HDHA Enhance antibody IgG2c Pathogenic mediated responses IgE class switching Enzymatic Enzymatic lgG/M IL-6 hydrolysis conversion IL-10

(S) OH (E) (S) OH COOH COOH (Z) (R) (Z) (E)(E) (E) (S) OH OH PDX Protectin D1/Neuroprotectin D1 (PD1/NPD1)

FIGURE 1 Biosynthesis and biological functions of SPM in B cells. (A) Biosynthetic pathway of PD1 and PDX. PD1 (10(R),17(S)-dihydroxy- 4Z,7Z,11E,13E,15Z,19Z-docosahexaenoic acid) produced enzymatically from the 16S, 17S epoxy protectin was confirmed by epoxide trap- ping experiments. PDX (10(S),17(S)-dihydroxy-4Z,7Z,11E,13Z,15E,19Z-docosahexaenoic acid) is a product of double dioxygenation pathway and occurs by sequential actions of 2 lipoxigenations. (B) SPM receptors on B cells and functions. LXA4 and RvD1 both activate ALX/FPR2 receptor present in B cells. SPMs enhance Ab-mediated responses and decreases pro-inflammatory cytokines

Resolution of inflammation: Role of B cells. Libreros S. J Leukoc Biol. 2019 Aug;106(2):235-239.

15 ..Egihe al. et English J.T.

Table 4 SPM and Eicosanoid Ocular Functions.

LM Main Bioactions Dose References Concentration within Human Tearsa

RvD1 Reduces vaso-obliteration and neovascularization 10 ng/day Connor et al. [13] 1.1 nM Reduction of inflammatory cytokines and mediators; reduction of PMN transmigration 50 nM; 50–200 nM Tian et al. [20] − − Block LTD₄ stimulated goblet cell secretions 10 9−10 8M Dartt et al. [11] − Regulation of histamine-stimulated goblet cell secretions 10 9 MLiet al. [22]; Hodges et al. [35,36] Block angiogenesis in inflamed cornea 100 ng/10 µL/mouse Reduction of inflammation in rat eye following uveitis 10–100-1000 ng/kg Hua et al. [14] Settimio et al. [17] RvE1 Reduces vaso-obliteration and neovascularization 10 ng/day Connor et al. [13] N/A Reduction of inflammatory cytokines and mediators; reduction of PMN transmigration 50 nM; 50–200 nM Tian et al. [20] − − Block LTD₄ stimulated goblet cell secretions 10 9−10 8M Dartt et al. [11] Block angiogenesis in inflamed cornea 100 ng/10 µL/mouse Jin et al. [15] Inhibition of corneal inflammation induced by LPS and bacteria 2 µg/2 µL PBS Lee et al. [16] PD1 Reduces vaso-obliteration and neovascularization 10 ng/day Connor et al. [13] 5.8 nM Increased corneal re-epithelialization and attenuation of thermal injury 1 µg Gronert et al. [10] Provides protection from oxidative-stress induced apoptotic damage in the retina 50 nM Mukherjee et al. [37] Restores corneal nerve integrity and function after surgery 100 ng, dropwise (topical) Cortina et al. [38] LXA₄ Decrease in inflammation, increase in wound healing; overall corneal & stroma protection 1 µg/eye in 10 µL PBS (mice) Kakazu et al. [19] 935 pM Effector T cell inhibition, regulatory T cell amplification, dry eye pathogenesis reduction 100 ng topically (3x/day), 1 µg systemically (daily) Gao et al. [12]; Hodges et al. [35,36] Reduction of VEGF-A and FLT4 expression and inflammatory angiogenesis 100 ng (3x daily) Leedom et al. [39] –

Attenuation of chemokine formation 1 1000 ng (3x daily) Biteman et al. [18] Prostaglandins, LeukotrienesandEssentialFattyAcids117(2017)17–27 Increased wound healing/re-epithelialization in cornea 1 µg (3x daily) Biteman et al. [18] AT-LXA₄ Suppression of VEGF-A induced hemoangiogenesis; control innate immune cell 100 ng/mouse He et al. [40] 1.1 nM infiltration Enhancement of endothelial cell wound closure and healing in cornea; 100 nM He et al. [40] Protection of corneal integrity during wound healing 100 nM He et al. [41]; Hodges et al. [35] − LTB₄ Stimulation of conjunctival goblet cell mucous secretion 10 10 M Dartt et al. [11] 16.4 nM PGE₂ Increase in inflammatory cytokines (IL-1β & IL-6) and intraoperative miosis following 25.6 pg/mL- 64.2 pg/m Wang et al. [42] 3.5 nM cataract surgery Signiﬁcantly higher presence in Dry Eye (DE) disease patient tears, compared to healthy 2.72 ± 3.42 ng/mL (average; n=23) Shim et al. [43] control tears Correlation with low tear osmolarity, meibomian gland plugging, and corneal staining 13.7 ± 3.1 pg/tear sample Walter et al. [24] − − PGF₂α Reduction in proliferation and adipogenesis 10 8−10 6 M Draman et al. [44] 3.8 nM Reduction in intraocular pressure (IOP) 50 µg/mL topical Lusky et al. [45]

a values from a representative donor

Identification and Profiling of Specialized Pro-Resolving Mediators in Human Tears by Lipid Mediator Metabolomics. English JT, Norris PC, Hodges RR, Dartt DA, Serhan CN. Prostaglandins Leukot Essent Fatty Acids. 2017 Feb;117:17-27.