- J. Delft van (1)
- J.H.M. Delft van (1)
- J. Ezendam (1)
- S. Gaj (1)
- R. Greuping (1)
- G. Groothuis (1)
- C. Hellfrisch (1)
- P. Hendriksen (1)
- D. Jennen (3)
- M.J.A. Jetten (1)
- J.C.S. Kleinjans (1)
- J. Kleinjans (2)
- T.M. Kok de (1)
- D. Kroese (1)
- F. Kuper (1)
- A. Lommen (2)
- H. Loveren van (1)
- M. Monshouwer (1)
- A.A.C.M. Peijnenburg (2)
- A. Ruiz Aracama (1)
- A. Ruiz-Aracama (1)
- F. Russel (1)
- L.G. Soeteman-Hernandez (1)
- E.P. Someren van (1)
- E. Someren van (1)
- R. Stierum (2)
- M. Tsamou (1)
Assuring safety without animal testing concept (ASAT). Integration of human disease data with in vitro data to improve toxicology testing
Stierum, R. ; Aarts, J.M.M.J.G. ; Boorsma, J. ; Bosgra, S. ; Caiment, F. ; Ezendam, J. ; Greuping, R. ; Hendriksen, P. ; Soeteman-Hernandez, L.G. ; Jennen, D. ; Kleinjans, J. ; Kroese, D. ; Kuper, F. ; Loveren, H. van; Monshouwer, M. ; Russel, F. ; Someren, E. van; Tsamou, M. ; Groothuis, G. - \ 2014
Toxicology Letters 229 (2014)suppl.10. - ISSN 0378-4274 - p. S4 - S21.
According to the Assuring Safety Without Animal Testing (ASAT) principle, risk assessment may ultimately become possible without the use of animals (Fentem et al., (2004). Altern. Lab. Anim. 32, 617–623). The ASAT concept takes human disease mechanisms as starting point and tries to define if activation of these mechanisms by chemical exposure in in vitro models can be used for toxicological risk assessment. The goal of the present research was to study if integration of public data, from human diseases and in vitro toxicology, is possible at the data level. Two human diseases, associated with chemical exposure, were included: human hepatocellular carcinoma and allergic contact dermatitis (ACD). Data were retrieved from expert knowledge and different online sources (e.g. GEO, ArrayExpress) and a Knowledge Base for storage and modelling of the data was established. Using the Knowledge Base for ACD, it was possible to discern sensitizing from non-sensitizing compounds, as defined by enrichment of clinically defined disease gene sets in in vitro genomics datasets. In addition, the strongest sensitizers most profoundly activated these gene sets. During the presentation, the Knowledge Base will be shown. In addition, the ongoing incorporation of (reverse) kinetic modelling to judge the relevance of in vitro concentrations, in relation to realistic in vivo exposure scenarios, will be illustrated. Finally, the expansion towards the development of data models for other disease areas (cholestasis) will be discussed.
'Omics analysis of low dose acetaminophen intake demonstrates novel response pathways in humans
Jetten, M.J.A. ; Gaj, S. ; Ruiz Aracama, A. ; Kok, T.M. de; Delft, J.H.M. van; Lommen, A. ; Someren, E.P. van; Jennen, D. ; Claessen, S.M. ; Peijnenburg, A.A.C.M. ; Stierum, R. ; Kleinjans, J.C.S. - \ 2012
Toxicology and Applied Pharmacology 259 (2012)3. - ISSN 0041-008X - p. 320 - 328.
induced liver-injury - gene-expression - induced hepatotoxicity - circulating micrornas - liquid-chromatography - potential biomarkers - toxicity - metabolomics - metabolites - paracetamol
Acetaminophen is the primary cause of acute liver toxicity in Europe/USA, which led the FDA to reconsider recommendations concerning safe acetaminophen dosage/use. Unfortunately, the current tests for liver toxicity are no ideal predictive markers for liver injury, i.e. they only measure acetaminophen exposure after profound liver toxicity has already occurred. Furthermore, these tests do not provide mechanistic information. Here, 'omics techniques (global analysis of metabolomic/gene-expression responses) may provide additional insight. To better understand acetaminophen-induced responses at low doses, we evaluated the effects of (sub-)therapeutic acetaminophen doses on metabolite formation and global gene-expression changes (including, for the first time, full-genome human miRNA expression changes) in blood/urine samples from healthy human volunteers. Many known and several new acetaminophen-metabolites were detected, in particular in relation to hepatotoxicity-linked, oxidative metabolism of acetaminophen. Transcriptomic changes indicated immune-modulating effects (2 g dose) and oxidative stress responses (4 g dose). For the first time, effects of acetaminophen on full-genome human miRNA expression have been considered and confirmed the findings on mRNA level. 'Omics techniques outperformed clinical chemistry tests and revealed novel response pathways to acetaminophen in humans. Although no definitive conclusion about potential immunotoxic effects of acetaminophen can be drawn from this study, there are clear indications that the immune system is triggered even after intake of low doses of acetaminophen. Also, oxidative stress-related gene responses, similar to those seen after high dose acetaminophen exposure, suggest the occurrence of possible pre-toxic effects of therapeutic acetaminophen doses. Possibly, these effects are related to dose-dependent increases in levels of hepatotoxicity-related metabolites.
An untargeted multi-technique metabolomics approach to studying intracellular metabolites of HepG2 cells exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin
Ruiz-Aracama, A. ; Peijnenburg, A.A.C.M. ; Kleinjans, J. ; Jennen, D. ; Delft, J. van; Hellfrisch, C. ; Lommen, A. - \ 2011
BMC Genomics 12 (2011). - ISSN 1471-2164
aryl-hydrocarbon receptor - gene-expression analysis - lipid-peroxidation - nongenotoxic carcinogens - protein glycosylation - liquid-chromatography - mass-spectrometry - human hepatocytes - oxidative stress - wasting syndrome
BackgroundIn vitro cell systems together with omics methods represent promising alternatives to conventional animal models for toxicity testing. Transcriptomic and proteomic approaches have been widely applied in vitro but relatively few studies have used metabolomics. Therefore, the goal of the present study was to develop an untargeted methodology for performing reproducible metabolomics on in vitro systems. The human liver cell line HepG2, and the well-known hepatotoxic and non-genotoxic carcinogen 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), were used as the in vitro model system and model toxicant, respectively.ResultsThe study focused on the analysis of intracellular metabolites using NMR, LC-MS and GC-MS, with emphasis on the reproducibility and repeatability of the data. State of the art pre-processing and alignment tools and multivariate statistics were used to detect significantly altered levels of metabolites after exposing HepG2 cells to TCDD. Several metabolites identified using databases, literature and LC-nanomate-Orbitrap analysis were affected by the treatment. The observed changes in metabolite levels are discussed in relation to the reported effects of TCDD.ConclusionsUntargeted profiling of the polar and apolar metabolites of in vitro cultured HepG2 cells is a valid approach to studying the effects of TCDD on the cell metabolome. The approach described in this research demonstrates that highly reproducible experiments and correct normalization of the datasets are essential for obtaining reliable results. The effects of TCDD on HepG2 cells reported herein are in agreement with previous studies and serve to validate the procedures used in the present work.