title: Metabolic Profiling as Well as Stable Isotope Assisted Metabolic and Proteomic Analysis of RAW 264.7 Macrophages Exposed to Ship Engine Aerosol Emissions: Different Effects of Heavy Fuel Oil and Refined Diesel Fuel.
abstract: Exposure to air pollution resulting from fossil fuel combustion has been linked to multiple short-term and long term health effects. In a previous study, exposure of lung epithelial cells to engine exhaust from heavy fuel oil (HFO) and diesel fuel (DF), two of the main fuels used in marine engines, led to an increased regulation of several pathways associated with adverse cellular effects, including pro-inflammatory pathways. In addition, DF exhaust exposure was shown to have a wider response on multiple cellular regulatory levels compared to HFO emissions, suggesting a potentially higher toxicity of DF emissions over HFO. In order to further understand these effects, as well as to validate these findings in another cell line, we investigated macrophages under the same conditions as a more inflammation-relevant model. An air-liquid interface aerosol exposure system was used to provide a more biologically relevant exposure system compared to submerged experiments, with cells exposed to either the complete aerosol (particle and gas phase), or the gas phase only (with particles filtered out). Data from cytotoxicity assays were integrated with metabolomics and proteomics analyses, including stable isotope-assisted metabolomics, in order to uncover pathways affected by combustion aerosol exposure in macrophages. Through this approach, we determined differing phenotypic effects associated with the different components of aerosol. The particle phase of diluted combustion aerosols was found to induce increased cell death in macrophages, while the gas phase was found more to affect the metabolic profile. In particular, a higher cytotoxicity of DF aerosol emission was observed in relation to the HFO aerosol. Furthermore, macrophage exposure to the gas phase of HFO leads to an induction of a pro-inflammatory metabolic and proteomic phenotype. These results validate the effects found in lung epithelial cells, confirming the role of inflammation and cellular stress in the response to combustion aerosols.
authors.affiliation: Luxembourg Centre for Systems Biomedicine 6, avenue du Swing, L-4362 Esch-sur-Alzette, Luxembourg.|HICE - Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health - Aerosols and Health, Neuherberg, Rostock, Munich, Karlsruhe, Berlin, Waldkirch, Germany; Kuopio, Finland; Cardiff, United Kingdom; Esch-Belval, Luxembourg.
authors.forename: Sean C
authors.initials: SC
authors.lastname: Sapcariu
authors.affiliation: Mass Spectrometry Core Unit, Max Delbrück Center for Molecular Medicine Berlin-Buch, Berlin, Germany.|HICE - Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health - Aerosols and Health, Neuherberg, Rostock, Munich, Karlsruhe, Berlin, Waldkirch, Germany; Kuopio, Finland; Cardiff, United Kingdom; Esch-Belval, Luxembourg.
authors.forename: Tamara
authors.initials: T
authors.lastname: Kanashova
authors.affiliation: Institute of Toxicology and Genetics (ITG), Karlsruhe Institute of Technology, Campus North, Karlsruhe, Germany.|Institute for Technical Chemistry (ITC), Karlsruhe Institute of Technology, Campus North, Karlsruhe, Germany.|HICE - Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health - Aerosols and Health, Neuherberg, Rostock, Munich, Karlsruhe, Berlin, Waldkirch, Germany; Kuopio, Finland; Cardiff, United Kingdom; Esch-Belval, Luxembourg.
authors.forename: Marco
authors.initials: M
authors.lastname: Dilger
authors.affiliation: Institute of Toxicology and Genetics (ITG), Karlsruhe Institute of Technology, Campus North, Karlsruhe, Germany.|HICE - Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health - Aerosols and Health, Neuherberg, Rostock, Munich, Karlsruhe, Berlin, Waldkirch, Germany; Kuopio, Finland; Cardiff, United Kingdom; Esch-Belval, Luxembourg.
authors.forename: Silvia
authors.initials: S
authors.lastname: Diabaté
authors.affiliation: Center of Allergy and Environment (ZAUM), Helmholtz Zentrum München and Technische Universität München, Munich, Germany.|CK-CARE, Christine Kühne Center for Allergy Research and Education, Davos, Switzerland.|HICE - Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health - Aerosols and Health, Neuherberg, Rostock, Munich, Karlsruhe, Berlin, Waldkirch, Germany; Kuopio, Finland; Cardiff, United Kingdom; Esch-Belval, Luxembourg.|German Center for Lung Research (DZL), Munich, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Oeder
authors.affiliation: Joint Mass Spectrometry Centre, Division of Analytical and Technical Chemistry, Institute of Chemistry, University Rostock, Rostock, Germany.|HICE - Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health - Aerosols and Health, Neuherberg, Rostock, Munich, Karlsruhe, Berlin, Waldkirch, Germany; Kuopio, Finland; Cardiff, United Kingdom; Esch-Belval, Luxembourg.
authors.forename: Johannes
authors.initials: J
authors.lastname: Passig
authors.affiliation: Joint Mass Spectrometry Centre, Division of Analytical and Technical Chemistry, Institute of Chemistry, University Rostock, Rostock, Germany.|HICE - Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health - Aerosols and Health, Neuherberg, Rostock, Munich, Karlsruhe, Berlin, Waldkirch, Germany; Kuopio, Finland; Cardiff, United Kingdom; Esch-Belval, Luxembourg.
authors.forename: Christian
authors.initials: C
authors.lastname: Radischat
authors.affiliation: Center of Allergy and Environment (ZAUM), Helmholtz Zentrum München and Technische Universität München, Munich, Germany.|CK-CARE, Christine Kühne Center for Allergy Research and Education, Davos, Switzerland.|HICE - Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health - Aerosols and Health, Neuherberg, Rostock, Munich, Karlsruhe, Berlin, Waldkirch, Germany; Kuopio, Finland; Cardiff, United Kingdom; Esch-Belval, Luxembourg.|German Center for Lung Research (DZL), Munich, Germany.
authors.forename: Jeroen
authors.initials: J
authors.lastname: Buters
authors.affiliation: University of Eastern Finland, Department of Environmental Science, P.O. Box 1627, FI-70211 Kuopio, Finland.|HICE - Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health - Aerosols and Health, Neuherberg, Rostock, Munich, Karlsruhe, Berlin, Waldkirch, Germany; Kuopio, Finland; Cardiff, United Kingdom; Esch-Belval, Luxembourg.
authors.forename: Olli
authors.initials: O
authors.lastname: Sippula
authors.affiliation: Joint Mass Spectrometry Centre, Division of Analytical and Technical Chemistry, Institute of Chemistry, University Rostock, Rostock, Germany.|Joint Mass Spectrometry Centre, CMA - Comprehensive Molecular Analytics, Helmholtz Zentrum München, Neuherberg, Germany.|HICE - Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health - Aerosols and Health, Neuherberg, Rostock, Munich, Karlsruhe, Berlin, Waldkirch, Germany; Kuopio, Finland; Cardiff, United Kingdom; Esch-Belval, Luxembourg.
authors.forename: Thorsten
authors.initials: T
authors.lastname: Streibel
authors.affiliation: Institute for Technical Chemistry (ITC), Karlsruhe Institute of Technology, Campus North, Karlsruhe, Germany.|HICE - Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health - Aerosols and Health, Neuherberg, Rostock, Munich, Karlsruhe, Berlin, Waldkirch, Germany; Kuopio, Finland; Cardiff, United Kingdom; Esch-Belval, Luxembourg.
authors.forename: Hanns-Rudolf
authors.initials: HR
authors.lastname: Paur
authors.affiliation: Institute for Technical Chemistry (ITC), Karlsruhe Institute of Technology, Campus North, Karlsruhe, Germany.|HICE - Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health - Aerosols and Health, Neuherberg, Rostock, Munich, Karlsruhe, Berlin, Waldkirch, Germany; Kuopio, Finland; Cardiff, United Kingdom; Esch-Belval, Luxembourg.
authors.forename: Christoph
authors.initials: C
authors.lastname: Schlager
authors.affiliation: Institute for Technical Chemistry (ITC), Karlsruhe Institute of Technology, Campus North, Karlsruhe, Germany.|HICE - Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health - Aerosols and Health, Neuherberg, Rostock, Munich, Karlsruhe, Berlin, Waldkirch, Germany; Kuopio, Finland; Cardiff, United Kingdom; Esch-Belval, Luxembourg.
authors.forename: Sonja
authors.initials: S
authors.lastname: Mülhopt
authors.affiliation: Chair of Piston Machines and Internal Combustion Engines, University Rostock, Rostock, Germany.|HICE - Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health - Aerosols and Health, Neuherberg, Rostock, Munich, Karlsruhe, Berlin, Waldkirch, Germany; Kuopio, Finland; Cardiff, United Kingdom; Esch-Belval, Luxembourg.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Stengel
authors.affiliation: Chair of Piston Machines and Internal Combustion Engines, University Rostock, Rostock, Germany.|HICE - Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health - Aerosols and Health, Neuherberg, Rostock, Munich, Karlsruhe, Berlin, Waldkirch, Germany; Kuopio, Finland; Cardiff, United Kingdom; Esch-Belval, Luxembourg.
authors.forename: Rom
authors.initials: R
authors.lastname: Rabe
authors.affiliation: Chair of Piston Machines and Internal Combustion Engines, University Rostock, Rostock, Germany.|HICE - Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health - Aerosols and Health, Neuherberg, Rostock, Munich, Karlsruhe, Berlin, Waldkirch, Germany; Kuopio, Finland; Cardiff, United Kingdom; Esch-Belval, Luxembourg.
authors.forename: Horst
authors.initials: H
authors.lastname: Harndorf
authors.affiliation: Vitrocell GmbH, Tübingen, Germany.|HICE - Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health - Aerosols and Health, Neuherberg, Rostock, Munich, Karlsruhe, Berlin, Waldkirch, Germany; Kuopio, Finland; Cardiff, United Kingdom; Esch-Belval, Luxembourg.
authors.forename: Tobias
authors.initials: T
authors.lastname: Krebs
authors.affiliation: Joint Mass Spectrometry Centre, CMA - Comprehensive Molecular Analytics, Helmholtz Zentrum München, Neuherberg, Germany.
authors.forename: Erwin
authors.initials: E
authors.lastname: Karg
authors.affiliation: Joint Mass Spectrometry Centre, CMA - Comprehensive Molecular Analytics, Helmholtz Zentrum München, Neuherberg, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Gröger
authors.affiliation: Institute of Toxicology and Genetics (ITG), Karlsruhe Institute of Technology, Campus North, Karlsruhe, Germany.|HICE - Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health - Aerosols and Health, Neuherberg, Rostock, Munich, Karlsruhe, Berlin, Waldkirch, Germany; Kuopio, Finland; Cardiff, United Kingdom; Esch-Belval, Luxembourg.
authors.forename: Carsten
authors.initials: C
authors.lastname: Weiss
authors.affiliation: Mass Spectrometry Core Unit, Max Delbrück Center for Molecular Medicine Berlin-Buch, Berlin, Germany.|HICE - Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health - Aerosols and Health, Neuherberg, Rostock, Munich, Karlsruhe, Berlin, Waldkirch, Germany; Kuopio, Finland; Cardiff, United Kingdom; Esch-Belval, Luxembourg.
authors.forename: Gunnar
authors.initials: G
authors.lastname: Dittmar
authors.affiliation: Luxembourg Centre for Systems Biomedicine 6, avenue du Swing, L-4362 Esch-sur-Alzette, Luxembourg.|HICE - Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health - Aerosols and Health, Neuherberg, Rostock, Munich, Karlsruhe, Berlin, Waldkirch, Germany; Kuopio, Finland; Cardiff, United Kingdom; Esch-Belval, Luxembourg.
authors.forename: Karsten
authors.initials: K
authors.lastname: Hiller
authors.affiliation: Joint Mass Spectrometry Centre, Division of Analytical and Technical Chemistry, Institute of Chemistry, University Rostock, Rostock, Germany.|Joint Mass Spectrometry Centre, CMA - Comprehensive Molecular Analytics, Helmholtz Zentrum München, Neuherberg, Germany.|HICE - Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health - Aerosols and Health, Neuherberg, Rostock, Munich, Karlsruhe, Berlin, Waldkirch, Germany; Kuopio, Finland; Cardiff, United Kingdom; Esch-Belval, Luxembourg.
authors.forename: Ralf
authors.initials: R
authors.lastname: Zimmermann
mesh_terms: D000818:Animals; D002460:Cell Line; D005648:Fuel Oils / Q000633:toxicity*; D005742:Gasoline / Q000633:toxicity*; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism; D055442:Metabolome / Q000187:drug effects*; D051379:Mice; D020543:Proteome / Q000187:drug effects*; D001335:Vehicle Emissions / Q000633:toxicity*
chemical_list: D005648:Fuel Oils; D005742:Gasoline; D020543:Proteome; D001335:Vehicle Emissions