title: Leukotriene E4 is a full functional agonist for human cysteinyl leukotriene type 1 receptor-dependent gene expression.
abstract: Leukotriene E4 (LTE4) the most stable of the cysteinyl leukotrienes (cysLTs) binds poorly to classical type 1 (CysLT1) and 2 (CysLT2) receptors although it induces potent responses in human airways in vivo, such as bronchoconstriction, airway hyperresponsiveness and inflammatory cell influx suggesting the presence of a novel receptor that preferentially responds to LTE4. To identify such a receptor two human mast cell lines, LAD2 and LUVA, were selected that differentially responded to LTE4 when analysed by intracellular signalling and gene expression. Comparative transcriptome analysis and recombinant gene overexpression experiments revealed CysLT1 as a receptor responsible for potent LTE4-induced response in LAD2 but not in LUVA cells, an observation confirmed further by gene knockdown and selective inhibitors. Lentiviral overexpression of CysLT1 in LUVA cells augmented intracellular calcium signalling induced by LTE4 but did not restore full agonist responses at the gene expression level. Our data support a model where both an increased expression of Gαq-coupled CysLT1, and sustained intracellular calcium mobilisation and extracellular signal-regulated kinase (Erk) activation, are required for LTE4-mediated regulation of gene expression in human cells. Our study shows for the first time that CysLT1 expression is critically important for responsiveness to LTE4 within a human cell system.
authors.affiliation: Division of Asthma, Allergy and Lung Biology, King's College London, UK.|MRC &Asthma UK Centre in Allergic Mechanisms of Asthma, UK.
authors.forename: Holly R
authors.initials: HR
authors.lastname: Foster
authors.affiliation: Division of Asthma, Allergy and Lung Biology, King's College London, UK.|MRC &Asthma UK Centre in Allergic Mechanisms of Asthma, UK.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Fuerst
authors.affiliation: Division of Asthma, Allergy and Lung Biology, King's College London, UK.|MRC &Asthma UK Centre in Allergic Mechanisms of Asthma, UK.
authors.forename: William
authors.initials: W
authors.lastname: Branchett
authors.affiliation: Division of Asthma, Allergy and Lung Biology, King's College London, UK.|MRC &Asthma UK Centre in Allergic Mechanisms of Asthma, UK.
authors.forename: Tak H
authors.initials: TH
authors.lastname: Lee
authors.affiliation: Division of Asthma, Allergy and Lung Biology, King's College London, UK.|MRC &Asthma UK Centre in Allergic Mechanisms of Asthma, UK.|NIHR Leicester Respiratory Biomedical Research Unit, Leicester Institute for Lung Health, Department of Infection, Immunity and Inflammation, University of Leicester, Leicester, UK.
authors.forename: David J
authors.initials: DJ
authors.lastname: Cousins
authors.affiliation: Division of Asthma, Allergy and Lung Biology, King's College London, UK.|MRC &Asthma UK Centre in Allergic Mechanisms of Asthma, UK.
authors.forename: Grzegorz
authors.initials: G
authors.lastname: Woszczek
mesh_terms: D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D015870:Gene Expression; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D017999:Leukotriene E4 / Q000378:metabolism* / Q000494:pharmacology; D008407:Mast Cells / Q000187:drug effects / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism; D018077:Receptors, Leukotriene / Q000819:agonists* / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D059467:Transcriptome
chemical_list: D034741:RNA, Small Interfering; D043562:Receptors, G-Protein-Coupled; D018077:Receptors, Leukotriene; D017999:Leukotriene E4; D048049:Extracellular Signal-Regulated MAP Kinases; C083169:leukotriene D4 receptor; D002118:Calcium
references.citation: Am Rev Respir Dis. 1990 Nov;142(5):1112-8
references.pmid: 2173457
references.citation: J Allergy Clin Immunol. 2015 May;135(5):1358-66.e1-11
references.pmid: 25441644
references.citation: J Biol Chem. 1995 Jun 23;270(25):15175-80
references.pmid: 7797501
references.citation: Nature. 1999 Jun 24;399(6738):789-93
references.pmid: 10391245
references.citation: J Allergy Clin Immunol. 2005 Feb;115(2):316-22
references.pmid: 15696087
references.citation: Am J Respir Cell Mol Biol. 2005 Dec;33(6):531-40
references.pmid: 16123393
references.citation: J Immunol. 2007 Apr 15;178(8):5262-70
references.pmid: 17404310
references.citation: N Engl J Med. 2007 Nov 1;357(18):1841-54
references.pmid: 17978293
references.citation: J Allergy Clin Immunol. 2008 Jan;121(1):215-221.e1
references.pmid: 18028998
references.citation: J Biol Chem. 2008 Jun 13;283(24):16477-87
references.pmid: 18411276
references.citation: Proc Natl Acad Sci U S A. 2008 Oct 28;105(43):16695-700
references.pmid: 18931305
references.citation: J Allergy Clin Immunol. 2009 Sep;124(3):417-21
references.pmid: 19482346
references.citation: J Exp Med. 2009 Oct 26;206(11):2543-55
references.pmid: 19822647
references.citation: J Allergy Clin Immunol. 2011 Mar;127(3):815-22.e1-5
references.pmid: 21281958
references.citation: J Allergy Clin Immunol. 2012 Apr;129(4):1136-42
references.pmid: 22391114
references.citation: Nucleic Acids Res. 2013 Jan;41(Database issue):D1083-8
references.pmid: 23087376
references.citation: PLoS One. 2013;8(3):e58305
references.pmid: 23472176
references.citation: J Biol Chem. 2013 Apr 19;288(16):10967-72
references.pmid: 23504326
references.citation: J Biol Chem. 2000 Sep 29;275(39):30531-6
references.pmid: 10851239
references.citation: Am J Respir Crit Care Med. 2001 Oct 15;164(8 Pt 1):1495-500
references.pmid: 11704602
references.citation: N Engl J Med. 2002 May 30;346(22):1699-705
references.pmid: 12037149
references.citation: N Engl J Med. 2002 Nov 7;347(19):1493-9
references.pmid: 12421891
references.citation: Leuk Res. 2003 Aug;27(8):677-82
references.pmid: 12801524
references.citation: Comp Biochem Physiol B. 1988;89(2):375-80
references.pmid: 3356141
references.citation: Lancet. 1993 Apr 17;341(8851):989-90
references.pmid: 8096945
references.citation: J Allergy Clin Immunol. 2013 Jul;132(1):205-13
references.pmid: 23688412
references.citation: Allergy. 2014 Nov;69(11):1531-9
references.pmid: 25041788
references.citation: Methods Mol Biol. 2015;1272:79-89
references.pmid: 25563178
references.citation: Eur Respir J. 1988 Dec;1(10):913-7
references.pmid: 2852123