title: Systems Toxicology Assessment of the Biological Impact of a Candidate Modified Risk Tobacco Product on Human Organotypic Oral Epithelial Cultures.
abstract: Cigarette smoke (CS) has been reported to increase predisposition to oral cancer and is also recognized as a risk factor for many conditions including periodontal diseases, gingivitis, and other benign mucosal disorders. Smoking cessation remains the most effective approach for minimizing the risk of smoking-related diseases. However, reduction of harmful constituents by heating rather than combusting tobacco, without modifying the amount of nicotine, is a promising new paradigm in harm reduction. In this study, we compared effects of exposure to aerosol derived from a candidate modified risk tobacco product, the tobacco heating system (THS) 2.2, with those of CS generated from the 3R4F reference cigarette. Human organotypic oral epithelial tissue cultures (EpiOral, MatTek Corporation) were exposed for 28 min to 3R4F CS or THS2.2 aerosol, both diluted with air to comparable nicotine concentrations (0.32 or 0.51 mg nicotine/L aerosol/CS for 3R4F and 0.31 or 0.46 mg/L for THS2.2). We also tested one higher concentration (1.09 mg/L) of THS2.2. A systems toxicology approach was employed combining cellular assays (i.e., cytotoxicity and cytochrome P450 activity assays), comprehensive molecular investigations of the buccal epithelial transcriptome (mRNA and miRNA) by means of computational network biology, measurements of secreted proinflammatory markers, and histopathological analysis. We observed that the impact of 3R4F CS was greater than THS2.2 aerosol in terms of cytotoxicity, morphological tissue alterations, and secretion of inflammatory mediators. Analysis of the transcriptomic changes in the exposed oral cultures revealed significant perturbations in various network models such as apoptosis, necroptosis, senescence, xenobiotic metabolism, oxidative stress, and nuclear factor (erythroid-derived 2)-like 2 (NFE2L2) signaling. The stress responses following THS2.2 aerosol exposure were markedly decreased, and the exposed cultures recovered more completely compared with those exposed to 3R4F CS.
authors.affiliation: Philip Morris International Research and Development , Quai Jeanrenaud 5, 2000 Neuchâtel, Switzerland.
authors.forename: Filippo
authors.initials: F
authors.lastname: Zanetti
authors.affiliation: Philip Morris International Research and Development , Quai Jeanrenaud 5, 2000 Neuchâtel, Switzerland.
authors.forename: Alain
authors.initials: A
authors.lastname: Sewer
authors.affiliation: Philip Morris International Research and Development , Quai Jeanrenaud 5, 2000 Neuchâtel, Switzerland.
authors.forename: Carole
authors.initials: C
authors.lastname: Mathis
authors.affiliation: Philip Morris International Research and Development , Quai Jeanrenaud 5, 2000 Neuchâtel, Switzerland.
authors.forename: Anita R
authors.initials: AR
authors.lastname: Iskandar
authors.affiliation: Philip Morris International Research and Development , Quai Jeanrenaud 5, 2000 Neuchâtel, Switzerland.
authors.forename: Radina
authors.initials: R
authors.lastname: Kostadinova
authors.affiliation: Biology Consultant , Max-Baermann-Str. 21, 51429 Bergisch Gladbach, Germany.
authors.forename: Walter K
authors.initials: WK
authors.lastname: Schlage
authors.affiliation: Philip Morris International Research and Development , Quai Jeanrenaud 5, 2000 Neuchâtel, Switzerland.
authors.forename: Patrice
authors.initials: P
authors.lastname: Leroy
authors.affiliation: Philip Morris International Research and Development , Quai Jeanrenaud 5, 2000 Neuchâtel, Switzerland.
authors.forename: Shoaib
authors.initials: S
authors.lastname: Majeed
authors.affiliation: Philip Morris International Research and Development , Quai Jeanrenaud 5, 2000 Neuchâtel, Switzerland.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Guedj
authors.affiliation: Philip Morris International Research and Development , Quai Jeanrenaud 5, 2000 Neuchâtel, Switzerland.
authors.forename: Keyur
authors.initials: K
authors.lastname: Trivedi
authors.affiliation: Philip Morris International Research and Development , Quai Jeanrenaud 5, 2000 Neuchâtel, Switzerland.
authors.forename: Florian
authors.initials: F
authors.lastname: Martin
authors.affiliation: Philip Morris International Research and Development , Quai Jeanrenaud 5, 2000 Neuchâtel, Switzerland.
authors.forename: Ashraf
authors.initials: A
authors.lastname: Elamin
authors.affiliation: Philip Morris International Research and Development , Quai Jeanrenaud 5, 2000 Neuchâtel, Switzerland.
authors.forename: Céline
authors.initials: C
authors.lastname: Merg
authors.affiliation: Philip Morris International Research and Development , Quai Jeanrenaud 5, 2000 Neuchâtel, Switzerland.
authors.forename: Nikolai V
authors.initials: NV
authors.lastname: Ivanov
authors.affiliation: Philip Morris International Research and Development , Quai Jeanrenaud 5, 2000 Neuchâtel, Switzerland.
authors.forename: Stefan
authors.initials: S
authors.lastname: Frentzel
authors.affiliation: Philip Morris International Research and Development , Quai Jeanrenaud 5, 2000 Neuchâtel, Switzerland.
authors.forename: Manuel C
authors.initials: MC
authors.lastname: Peitsch
authors.affiliation: Philip Morris International Research and Development , Quai Jeanrenaud 5, 2000 Neuchâtel, Switzerland.
authors.forename: Julia
authors.initials: J
authors.lastname: Hoeng
mesh_terms: D004781:Environmental Exposure; D006801:Humans; D035683:MicroRNAs / Q000378:metabolism; D009061:Mouth Mucosa / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D014116:Toxicology; D014026:Nicotiana
chemical_list: D035683:MicroRNAs