Toxicity and metabolism preclinical studies Unit
This Unit provides hepatic and other tissue cell models to investigate drug metabolism, as well as the tests to assess xenobiotic-induced toxicity and the molecular mechanisms involved to the researchers and the pharmaceutical, cosmetic and chemical industry.
FACILITIES AND EQUIPMENT
• Cell culture facilities (4 rooms).
• Molecular biology equipments.
• Microplate readers (absorbance, fluorescence, luminiscence).
• Fluorescence microscopy.
• Scan^R (Olympus) high-content screening (HCS) station by image analysis.
• Chromatographic equipments (HPLC-MS/MS).
SERVICES AND EXPERTISSE
- Primary culture of hepatocytes from various animal species
- Human hepatoma cell lines and of several animal species
- Genetically engineered cellular models expressing genes of interest (permanent or transient).
- Other cellular models (fibroblasts, queratinocytes, cell lines, etc)
- Cocktail of specific substrates of the cytochrome P450 and conjugating enzymes to determine the metabolic capacity of the cells and drug induction and inhibition studies of cytochrome P450 enzymes.
- Adenovirus expression vectors (transcription factors, nuclear receptors, transporters, cytochrome P450 and conjugation enzymes, etc.)
- Development of customized cell models expressing transiently one or several genes of interest by transfection with adenovirus expression vectors.
- Metabonomics: Screening model to predict and classify the potential toxicity of new compounds by identifying cell metabolites as specific biomarkers of each toxicity mechanism.
- High-content image analysis (HCA) techniques of for multiparametric study of the mechanisms of toxicity of xenobiotics. Combinations of compatible fluorescent probes to assess several mechanisms simultaneously are used.
Services for metabolism research and xenobiotics toxicity
Preclinical study of toxicity
- Evaluation of basal/hepatic toxicity (MTT cytotoxicity test, Neutral red, etc.)
- Metabolism-dependent toxicity: Bioactivation/detoxification
- Effects of xenobiotics on specific liver functions (ureogenesis, gluconeogenesis, plasma protein synthesis, glycogen, GSH, etc).
- Molecular mechanism of toxicity (oxidative stress, mitochondrial toxicity, intracelullar Ca+2 levels, lipid peroxidation, etc.) using high-content image analysis techniques.
- Chronic toxicity (assay to detect compounds inducing steatosis) using high-content image analysis and transcriptomic techniques.
Preclinical study of metabolism
- Metabolic stability/metabolism rates/clearance.
- Metabolic profile / metabolite identification (conjugated and non-conjugated) in collaboration with the Analytical Unit.
- Identification of cytochrome P450 isoenzymes involved in the metabolism: contribution of each isozyme/contribution of isoenzymes with genetic polymorphism.
- Induction/inhibition of cytochrome P450 isoenzymes: Drug-drug interaction.
- Selection of the specie closest to man for preclinical studies in animals, by the comparative study of drug metabolism in cultured hepatocytes from several animal species and human.