title: Macrophage epoxygenase determines a profibrotic transcriptome signature.
abstract: Epoxygenases belong to the cytochrome P450 family. They generate epoxyeicosatrienoic acids, which are known to have anti-inflammatory effects, but little is known about their role in macrophage function. By high-throughput sequencing of RNA in primary macrophages derived from rodents and humans, we establish the relative expression of epoxygenases in these cells. Zinc-finger nuclease-mediated targeted gene deletion of the major rat macrophage epoxygenase Cyp2j4 (ortholog of human CYP2J2) resulted in reduced epoxyeicosatrienoic acid synthesis. Cyp2j4(-/-) macrophages have relatively increased peroxisome proliferator-activated receptor-γ levels and show a profibrotic transcriptome, displaying overexpression of a specific subset of genes (260 transcripts) primarily involved in extracellular matrix, with fibronectin being the most abundantly expressed transcript. Fibronectin expression is under the control of epoxygenase activity in human and rat primary macrophages. In keeping with the in vitro findings, Cyp2j4(-/-) rats show upregulation of type I collagen following unilateral ureter obstruction of the kidney, and quantitative proteomics analysis (liquid chromatography-tandem mass spectrometry) showed increased renal type I collagen and fibronectin protein abundance resulting from experimentally induced crescentic glomerulonephritis in these rats. Taken together, these results identify the rat epoxygenase Cyp2j4 as a determinant of a profibrotic macrophage transcriptome that could have implications in various inflammatory conditions, depending on macrophage function.
authors.affiliation: Centre for Complement and Inflammation Research (CCIR), Imperial College London, W12 0NN, London, UK.
authors.forename: Jacques
authors.initials: J
authors.lastname: Behmoaras
authors.affiliation: Physiological Genomics and Medicine, MRC Clinical Sciences Centre, Imperial College London, W12 0NN, UK.
authors.forename: Ana Garcia
authors.initials: AG
authors.lastname: Diaz
authors.affiliation: Physiological Genomics and Medicine, MRC Clinical Sciences Centre, Imperial College London, W12 0NN, UK.
authors.forename: Lara
authors.initials: L
authors.lastname: Venda
authors.affiliation: Centre for Complement and Inflammation Research (CCIR), Imperial College London, W12 0NN, London, UK.
authors.forename: Jeong-Hun
authors.initials: JH
authors.lastname: Ko
authors.affiliation: Integrative Genomics and Medicine, MRC Clinical Sciences Centre, Imperial College London, W12 0NN, UK and Duke-NUS Graduate Medical School Singapore. 8 College Road, 169857 Singapore, Republic of Singapore.
authors.forename: Prashant
authors.initials: P
authors.lastname: Srivastava
authors.affiliation: Biological Mass Spectrometry and Proteomics Laboratory, MRC Clinical Sciences Centre, Imperial College London, W12 0NN, UK.
authors.forename: Alex
authors.initials: A
authors.lastname: Montoya
authors.affiliation: Biological Mass Spectrometry and Proteomics Laboratory, MRC Clinical Sciences Centre, Imperial College London, W12 0NN, UK.
authors.forename: Peter
authors.initials: P
authors.lastname: Faull
authors.affiliation: ES Cell and Transgenics Facility, MRC Clinical Sciences Centre, Imperial College London, W12 0NN, UK.
authors.forename: Zoe
authors.initials: Z
authors.lastname: Webster
authors.affiliation: ES Cell and Transgenics Facility, MRC Clinical Sciences Centre, Imperial College London, W12 0NN, UK.
authors.forename: Ben
authors.initials: B
authors.lastname: Moyon
authors.affiliation: Renal Section, Department of Medicine, Imperial College London, Hammersmith Campus, London, UK.
authors.forename: Charles D
authors.initials: CD
authors.lastname: Pusey
authors.affiliation: Centre for Rheumatology & Connective Tissue Diseases, University College London Medical School, London, UK.
authors.forename: David J
authors.initials: DJ
authors.lastname: Abraham
authors.affiliation: Integrative Genomics and Medicine, MRC Clinical Sciences Centre, Imperial College London, W12 0NN, UK and Duke-NUS Graduate Medical School Singapore. 8 College Road, 169857 Singapore, Republic of Singapore.
authors.forename: Enrico
authors.initials: E
authors.lastname: Petretto
authors.affiliation: Centre for Complement and Inflammation Research (CCIR), Imperial College London, W12 0NN, London, UK.
authors.forename: Terence H
authors.initials: TH
authors.lastname: Cook
authors.affiliation: Physiological Genomics and Medicine, MRC Clinical Sciences Centre, Imperial College London, W12 0NN, UK.|Institute of Genetics & Molecular Medicine, University of Edinburgh, EH4 2XU, UK.
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Aitman
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002853:Chromatography, Liquid; D000090762:Cytochrome P-450 CYP2J2; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D000072467:Cytochrome P450 Family 2; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay; D005355:Fibrosis / Q000201:enzymology* / Q000235:genetics*; D055786:Gene Knockout Techniques; D005921:Glomerulonephritis / Q000201:enzymology / Q000235:genetics; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008264:Macrophages / Q000201:enzymology*; D008297:Male; D034622:RNA Interference; D051381:Rats; D011921:Rats, Inbred WKY; D060888:Real-Time Polymerase Chain Reaction; D053719:Tandem Mass Spectrometry; D059467:Transcriptome
chemical_list: C000717350:CYP2J2 protein, human; D003577:Cytochrome P-450 Enzyme System; C112496:Cyp2j4 protein, rat; D000090762:Cytochrome P-450 CYP2J2; D000072467:Cytochrome P450 Family 2
references.citation: J Cell Physiol. 2008 Jun;215(3):771-81
references.pmid: 18181170
references.citation: Kidney Int. 1990 Jan;37(1):94-9
references.pmid: 2105407
references.citation: Life Sci. 2008 Aug 29;83(9-10):339-45
references.pmid: 18675280
references.citation: Pharmacol Rev. 2014 Oct;66(4):1106-40
references.pmid: 25244930
references.citation: Nat Rev Drug Discov. 2009 Oct;8(10):794-805
references.pmid: 19794443
references.citation: N Engl J Med. 2007 Nov 1;357(18):1841-54
references.pmid: 17978293
references.citation: Physiol Rev. 2012 Jan;92(1):101-30
references.pmid: 22298653
references.citation: F1000Prime Rep. 2014 Mar 03;6:13
references.pmid: 24669294
references.citation: Proc Natl Acad Sci U S A. 1981 Nov;78(11):7147-51
references.pmid: 6947279
references.citation: Genes Dev. 2002 Jan 1;16(1):22-6
references.pmid: 11782441
references.citation: Bioinformatics. 2010 Jan 1;26(1):139-40
references.pmid: 19910308
references.citation: Hypertension. 2014 Dec;64(6):1352-61
references.pmid: 25245389
references.citation: J Immunol. 1998 Sep 1;161(5):2441-8
references.pmid: 9725242
references.citation: Proc Natl Acad Sci U S A. 2013 Aug 13;110(33):13528-33
references.pmid: 23898174
references.citation: Blood. 2012 Oct 11;120(15):e60-72
references.pmid: 22904297
references.citation: Proc Natl Acad Sci U S A. 2005 Nov 15;102(46):16747-52
references.pmid: 16267130
references.citation: Nat Rev Immunol. 2011 Nov;11(11):723-37
references.pmid: 21997792
references.citation: Am J Physiol Renal Physiol. 2009 Sep;297(3):F740-8
references.pmid: 19553349
references.citation: Cell. 2005 Dec 16;123(6):993-9
references.pmid: 16360030
references.citation: J Biol Chem. 2001 Sep 28;276(39):36059-62
references.pmid: 11451964
references.citation: J Lipid Res. 2013 May;54(5):1448-56
references.pmid: 23446230
references.citation: BMC Genomics. 2013;14:92
references.pmid: 23398888
references.citation: Genes Dev. 2008 Nov 1;22(21):2941-52
references.pmid: 18981473
references.citation: J Clin Invest. 1987 Dec;80(6):1720-7
references.pmid: 3680524
references.citation: J Immunol. 2008 Apr 15;180(8):5707-19
references.pmid: 18390756
references.citation: FASEB J. 2014 Jun;28(6):2538-50
references.pmid: 24571919
references.citation: Hypertension. 2013 Nov;62(5):905-13
references.pmid: 23980070
references.citation: Nat Rev Nephrol. 2014 Sep;10(9):493-503
references.pmid: 24981817
references.citation: Cell Rep. 2014 Aug 21;8(4):1210-24
references.pmid: 25131209
references.citation: J Biol Chem. 1996 Feb 16;271(7):3460-8
references.pmid: 8631948
references.citation: Endocrinology. 2013 Aug;154(8):2843-56
references.pmid: 23696562
references.citation: J Vis Exp. 2010;(44). pii: 2033. doi: 10.3791/2033
references.pmid: 20972406
references.citation: FASEB J. 2004 May;18(7):816-27
references.pmid: 15117886
references.citation: FASEB J. 2009 Sep;23(9):2968-77
references.pmid: 19395477
references.citation: Mol Pharmacol. 2014 Jan;85(1):105-15
references.pmid: 24145329
references.citation: Hum Gene Ther. 2012 Jul;23(7):688-99
references.pmid: 22260463
references.citation: J Leukoc Biol. 2008 Sep;84(3):623-30
references.pmid: 18467655
references.citation: Mol Cell Proteomics. 2015 Mar;14(3):484-98
references.pmid: 25532521
references.citation: Proc Natl Acad Sci U S A. 2005 Feb 8;102(6):2186-91
references.pmid: 15684051
references.citation: J Am Soc Nephrol. 2010 Jul;21(7):1136-44
references.pmid: 20488952
references.citation: Arthritis Rheum. 2004 Apr;50(4):1305-18
references.pmid: 15077315
references.citation: Nat Genet. 2008 May;40(5):553-9
references.pmid: 18443593
references.citation: Basic Res Cardiol. 2013 Jan;108(1):319
references.pmid: 23224081
references.citation: J Immunol. 2006 Nov 15;177(10):7303-11
references.pmid: 17082649
references.citation: Nephrol Dial Transplant. 1999 Jul;14(7):1658-66
references.pmid: 10435873
references.citation: Pharmacol Ther. 2014 Nov;144(2):134-61
references.pmid: 24882266
references.citation: PLoS Genet. 2013 Nov;9(11):e1003950
references.pmid: 24278032
references.citation: PLoS One. 2013;8(9):e75107
references.pmid: 24058654
references.citation: Dis Model Mech. 2013 Nov;6(6):1477-86
references.pmid: 24046355
references.citation: Science. 1999 Aug 20;285(5431):1276-9
references.pmid: 10455056
references.citation: FASEB J. 2011 Feb;25(2):703-13
references.pmid: 21059750