title: Quantitative proteomic analysis of hepatocyte-secreted extracellular vesicles reveals candidate markers for liver toxicity.
abstract: Extracellular vesicles have created great interest as possible source of biomarkers for different biological processes and diseases. Although the biological function of these vesicles is not fully understood, it is clear that they participate in the removal of unnecessary cellular material and act as carriers of various macromolecules and signals between the cells. In this report, we analyzed the proteome of extracellular vesicles secreted by primary hepatocytes. We used one- and two-dimensional liquid chromatography combined with data-independent mass spectrometry. Employing label-free quantitative proteomics, we detected significant changes in vesicle protein expression levels in this in vitro model after exposure to well-known liver toxins (galactosamine and Escherichia coli-derived lipopolysaccharide). The results allowed us to identify candidate markers for liver injury. We validated a number of these markers in vivo, providing the basis for the development of novel methods to evaluate drug toxicity. This report strongly supports the application of proteomics in the study of extracellular vesicles released by well-controlled in vitro cellular systems. Analysis of such systems should help to identify specific markers for various biological processes and pathological conditions.
  
  
  CONCLUSIONS
  Identification of low invasive candidate marker for hepatotoxicity. Support to apply proteomics in the study of extracellular vesicles released by well-controlled in vitro cellular systems to identify low invasive markers for diseases.
authors.affiliation: Proteomics Platform, CIC bioGUNE, CIBERehd, ProteoRed-ISCIII, Derio, Spain.
authors.forename: Eva
authors.initials: E
authors.lastname: Rodríguez-Suárez
authors.affiliation: Metabolomics Unit, CIC bioGUNE, CIBERehd, Derio, Spain.
authors.forename: Esperanza
authors.initials: E
authors.lastname: Gonzalez
authors.affiliation: Waters Corporation, MS Technologies Center, Manchester, United Kingdom.
authors.forename: Chris
authors.initials: C
authors.lastname: Hughes
authors.affiliation: Metabolomics Unit, CIC bioGUNE, CIBERehd, Derio, Spain.
authors.forename: Javier
authors.initials: J
authors.lastname: Conde-Vancells
authors.affiliation: Waters Corporation, Milan, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Rudella
authors.affiliation: Metabolomics Unit, CIC bioGUNE, CIBERehd, Derio, Spain.
authors.forename: Felix
authors.initials: F
authors.lastname: Royo
authors.affiliation: Metabolomics Unit, CIC bioGUNE, CIBERehd, Derio, Spain.
authors.forename: Laura
authors.initials: L
authors.lastname: Palomo
authors.affiliation: Proteomics Platform, CIC bioGUNE, CIBERehd, ProteoRed-ISCIII, Derio, Spain.
authors.forename: Felix
authors.initials: F
authors.lastname: Elortza
authors.affiliation: USC Research Center for Liver Diseases, Division of Gastrointestinal and Liver Diseases, Keck School of Medicine, University of Southern California, Los Angeles, CA, United States.
authors.forename: Shelly C
authors.initials: SC
authors.lastname: Lu
authors.affiliation: Metabolomics Unit, CIC bioGUNE, CIBERehd, Derio, Spain.
authors.forename: Jose M
authors.initials: JM
authors.lastname: Mato
authors.affiliation: Waters Corporation, MS Technologies Center, Manchester, United Kingdom.
authors.forename: Johannes P C
authors.initials: JP
authors.lastname: Vissers
authors.affiliation: Metabolomics Unit, CIC bioGUNE, CIBERehd, Derio, Spain; IKERBASQUE, Basque Foundation for Science, Bilbao, Spain. Electronic address: jfalcon@cicbiogune.es.
authors.forename: Juan M
authors.initials: JM
authors.lastname: Falcón-Pérez
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000032:analysis*; D056486:Chemical and Drug Induced Liver Injury / Q000378:metabolism*; D055354:Exosomes / Q000737:chemistry; D005688:Galactosamine / Q000633:toxicity; D022781:Hepatocytes / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000633:toxicity; D008099:Liver / Q000737:chemistry / Q000187:drug effects; D008297:Male; D013058:Mass Spectrometry; D040901:Proteomics / Q000379:methods; D017207:Rats, Sprague-Dawley; D022142:Secretory Vesicles / Q000737:chemistry*
chemical_list: D015415:Biomarkers; D008070:Lipopolysaccharides; D005688:Galactosamine
keywords: Exosomes; Hepatocytes; LC–MS; Label-free quantitation; Liver; Microvesicles
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