title: Combined Proteome and Eicosanoid Profiling Approach for Revealing Implications of Human Fibroblasts in Chronic Inflammation.
abstract: During inflammation, proteins and lipids act in a concerted fashion, calling for combined analyses. Fibroblasts are powerful mediators of chronic inflammation. However, little is known about eicosanoid formation by human fibroblasts. The aim of this study was to analyze the formation of the most relevant inflammation mediators including proteins and lipids in human fibroblasts upon inflammatory stimulation and subsequent treatment with dexamethasone, a powerful antiphlogistic drug. Label-free quantification was applied for proteome profiling, while an in-house established data-dependent analysis method based on high-resolution mass spectrometry was applied for eicosadomics. Furthermore, a set of 188 metabolites was determined by targeted analysis. The secretion of 40 proteins including cytokines, proteases, and other inflammation agonists as well as 14 proinflammatory and nine anti-inflammatory eicosanoids was found significantly induced, while several acylcarnithins and sphingomyelins were found significantly downregulated upon inflammatory stimulation. Treatment with dexamethasone downregulated most cytokines and proteases, abrogated the formation of pro- but also anti-inflammatory eicosanoids, and restored normal levels of acylcarnithins but not of sphingomyelins. In addition, the chemokines CXCL1, CXCL5, CXCL6, and complement C3, known to contribute to chronic inflammation, were not counter-regulated by dexamethasone. Similar findings were obtained with human mesenchymal stem cells, and results were confirmed by targeted analysis with multiple reaction monitoring. Comparative proteome profiling regarding other cells demonstrated cell-type-specific synthesis of, among others, eicosanoid-forming enzymes as well as relevant transcription factors, allowing us to better understand cell-type-specific regulation of inflammation mediators and shedding new light on the role of fibroblasts in chronic inflammation.
authors.affiliation: Department of Analytical Chemistry, Faculty of Chemistry, University of Vienna , 1090 Vienna, Austria.
authors.forename: Ammar
authors.initials: A
authors.lastname: Tahir
authors.affiliation: Department of Analytical Chemistry, Faculty of Chemistry, University of Vienna , 1090 Vienna, Austria.
authors.forename: Andrea
authors.initials: A
authors.lastname: Bileck
authors.affiliation: Department of Analytical Chemistry, Faculty of Chemistry, University of Vienna , 1090 Vienna, Austria.
authors.forename: Besnik
authors.initials: B
authors.lastname: Muqaku
authors.affiliation: Department of Analytical Chemistry, Faculty of Chemistry, University of Vienna , 1090 Vienna, Austria.
authors.forename: Laura
authors.initials: L
authors.lastname: Niederstaetter
authors.affiliation: Department of Analytical Chemistry, Faculty of Chemistry, University of Vienna , 1090 Vienna, Austria.
authors.forename: Dominique
authors.initials: D
authors.lastname: Kreutz
authors.affiliation: Department of Analytical Chemistry, Faculty of Chemistry, University of Vienna , 1090 Vienna, Austria.
authors.forename: Rupert L
authors.initials: RL
authors.lastname: Mayer
authors.affiliation: Department of Analytical Chemistry, Faculty of Chemistry, University of Vienna , 1090 Vienna, Austria.
authors.forename: Denise
authors.initials: D
authors.lastname: Wolrab
authors.affiliation: Department of Analytical Chemistry, Faculty of Chemistry, University of Vienna , 1090 Vienna, Austria.
authors.forename: Samuel M
authors.identifier: 0000-0002-8930-4574
authors.initials: SM
authors.lastname: Meier
authors.affiliation: Department of Analytical Chemistry, Faculty of Chemistry, University of Vienna , 1090 Vienna, Austria.
authors.forename: Astrid
authors.identifier: 0000-0002-2217-5800
authors.initials: A
authors.lastname: Slany
authors.affiliation: Department of Analytical Chemistry, Faculty of Chemistry, University of Vienna , 1090 Vienna, Austria.
authors.forename: Christopher
authors.identifier: 0000-0003-4964-0642
authors.initials: C
authors.lastname: Gerner
mesh_terms: D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D002478:Cells, Cultured; D018925:Chemokines / Q000378:metabolism; D002853:Chromatography, Liquid / Q000379:methods; D002908:Chronic Disease; D016207:Cytokines / Q000378:metabolism; D003907:Dexamethasone / Q000494:pharmacology; D015777:Eicosanoids / Q000378:metabolism*; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007249:Inflammation / Q000097:blood / Q000378:metabolism* / Q000473:pathology; D018836:Inflammation Mediators / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods; D055432:Metabolomics; D020543:Proteome
chemical_list: D000893:Anti-Inflammatory Agents; D018925:Chemokines; D016207:Cytokines; D015777:Eicosanoids; D018836:Inflammation Mediators; D020543:Proteome; D003907:Dexamethasone