title: Impact of a synthetic cannabinoid (CP-47,497-C8) on protein expression in human cells: evidence for induction of inflammation and DNA damage.
abstract: Synthetic cannabinoids (SCs) are marketed worldwide as legal surrogates for marihuana. In order to predict potential health effects in consumers and to elucidate the underlying mechanisms of action, we investigated the impact of a representative of the cyclohexylphenols, CP47,497-C8, which binds to both cannabinoid receptors, on protein expression patterns, genomic stability and on induction of inflammatory cytokines in human lymphocytes. After treatment of the cells with the drug, we found pronounced up-regulation of a variety of enzymes in nuclear extracts which are involved in lipid metabolism and inflammatory signaling; some of the identified proteins are also involved in the endogenous synthesis of endocannabinoids. The assumption that the drug causes inflammation is further supported by results obtained in additional experiments with cytosols of LPS-stimulated lymphocytes which showed that the SC induces pro-inflammatory cytokines (IL12p40 and IL-6) as well as TNF-α. Furthermore, the proteome analyses revealed that the drug causes down-regulation of proteins which are involved in DNA repair. This observation provides an explanation for the formation of comets which was seen in single-cell gel electrophoresis assays and for the induction of micronuclei (which reflect structural and numerical chromosomal aberrations) by the drug. These effects were seen in experiments with human lymphocytes which were conducted under identical conditions as the proteome analysis. Taken together, the present findings indicate that the drug (and possibly other structurally related SCs) may cause DNA damage and inflammation in directly exposed cells of consumers.
authors.affiliation: Faculty of Chemistry, Institute of Analytical Chemistry, University of Vienna, Währingerstr. 38, 1090, Vienna, Austria.
authors.forename: Andrea
authors.initials: A
authors.lastname: Bileck
authors.affiliation: Department of Internal Medicine 1, Comprehensive Cancer Center, Institute of Cancer Research, Medical University of Vienna, Borschkegasse 8A, 1090, Vienna, Austria.
authors.forename: Franziska
authors.initials: F
authors.lastname: Ferk
authors.affiliation: Department of Internal Medicine 1, Comprehensive Cancer Center, Institute of Cancer Research, Medical University of Vienna, Borschkegasse 8A, 1090, Vienna, Austria.
authors.forename: Halh
authors.initials: H
authors.lastname: Al-Serori
authors.affiliation: Department of Internal Medicine 1, Comprehensive Cancer Center, Institute of Cancer Research, Medical University of Vienna, Borschkegasse 8A, 1090, Vienna, Austria.
authors.forename: Verena J
authors.initials: VJ
authors.lastname: Koller
authors.affiliation: Faculty of Chemistry, Institute of Analytical Chemistry, University of Vienna, Währingerstr. 38, 1090, Vienna, Austria.
authors.forename: Besnik
authors.initials: B
authors.lastname: Muqaku
authors.affiliation: Department of Nutritional Sciences, University of Vienna, UZA 2/2D541, Althanstrasse 14, 1090, Vienna, Austria.
authors.forename: Alexander
authors.initials: A
authors.lastname: Haslberger
authors.affiliation: Institute of Forensic Medicine, University Medical Center Freiburg, Albertstraße 9, 79104, Freiburg, Germany.
authors.forename: Volker
authors.initials: V
authors.lastname: Auwärter
authors.affiliation: Faculty of Chemistry, Institute of Analytical Chemistry, University of Vienna, Währingerstr. 38, 1090, Vienna, Austria.
authors.forename: Christopher
authors.initials: C
authors.lastname: Gerner
authors.affiliation: Department of Internal Medicine 1, Comprehensive Cancer Center, Institute of Cancer Research, Medical University of Vienna, Borschkegasse 8A, 1090, Vienna, Austria. siegfried.knasmueller@meduniwien.ac.at.
authors.forename: Siegfried
authors.initials: S
authors.lastname: Knasmüller
mesh_terms: D000328:Adult; D002186:Cannabinoids / Q000633:toxicity*; D002478:Cells, Cultured; D002853:Chromatography, Liquid; D020552:Comet Assay; D003511:Cyclohexanols / Q000633:toxicity*; D016207:Cytokines / Q000096:biosynthesis*; D004249:DNA Damage; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008213:Lymphocyte Activation / Q000187:drug effects; D008214:Lymphocytes / Q000187:drug effects* / Q000276:immunology; D008297:Male; D013058:Mass Spectrometry; D009687:Nuclear Proteins / Q000096:biosynthesis*; D010636:Phenols / Q000633:toxicity*; D040081:Protein Array Analysis; D014176:Protein Biosynthesis / Q000187:drug effects / Q000276:immunology; D013347:Subcellular Fractions
chemical_list: C572491:3-(2-hydroxy-4-(2-methylnonan-2-yl)phenyl)cyclohexan-1-ol; D002186:Cannabinoids; D003511:Cyclohexanols; D016207:Cytokines; D008070:Lipopolysaccharides; D009687:Nuclear Proteins; D010636:Phenols
keywords: Comet assay; Lymphocytes; Proteomics; Synthetic cannabinoid