- Email: [email protected]
In vitro carcinogenicity of perfluorooctane sulfonate (PFOS) on Syrian hamster embryo (SHE) cells
Abstracts / Toxicology Letters 205S (2011) S36–S59
progresses. The LTKB-BD has good representation of known drugs in terms of chemical structure, therapeutic categories, and daily dosage. The LTKB-BD has the potential to serve as a benchmark dataset for the community-wide effort to evaluate the performance characteristics of existing DILI biomarkers and to develop novel DILI biomarkers using emerging molecular technologies. doi:10.1016/j.toxlet.2011.05.217
OS11 In Vitro Toxicology
OS11-1 Evaluation of drug-induced toxicity in isolated mouse liver mitochondria as a predictive model of hepatotoxicity risks in human M. Porceddu 1 , N. Buron 1 , C. Roussel 1 , B. Fromenty 2 , A. Borgne-Sanchez 1,∗ 1
MITOLOGICS SAS, Paris, France, 2 U991, INSERM, Rennes, France
Purpose: Mitochondrial dysfunction is a major mechanism of drug-induced liver injury (DILI), which can be severe and even fatal. DILI can thus lead to the interruption of clinical trials, or drug withdrawal after marketing. As the financial losses can be important for pharmaceutical companies, drug-induced mitochondrial dysfunction and DILI should be detected at early development stages. Methods: To facilitate this detection, we propose a high-throughput screening platform on isolated mouse liver mitochondria which allows the measurement of several parameters pertaining to mitochondrial function and membrane integrity. Indeed, these multiparametric assays measure: (1) global mitochondrial membrane permeabilization, (2) transmembrane potential, (3) mitochondrial outer membrane permeabilization and (4) respiration in isolated mouse liver mitochondria. Results: The screening of 100 drugs known to be hepatotoxic (Alpidem, Lumiracoxib, Troglitazone. . .) or not, allowed us to establish a strong correlation between in vitro mitochondrio-toxicity and DILI in human. A robust statistical analysis demonstrates the high predictivity level of this screening platform. doi:10.1016/j.toxlet.2011.05.219
OS11-2 Optimization of an in vitro model to study environmental pollutant effects on pancreatic -cell function and evaluation of the mode of action of phthalates T. Hectors ∗ , C. Vanparys, A. Pereira-Fernandes, R. Blust Ecophysiology, Biochemistry and Toxicology, University of Antwerp, Antwerp, Belgium
-cell failure is the triggering factor for the progression of prediabetic states to type 2 diabetes, there is a great need to focus on the impact of chemicals on -cells. Therefore, we investigated if the pancreatic -cell line INS-1 832/13 can be used as a system for the evaluation of compound effects and we optimized exposure conditions using pharmaceutical compounds. Secondly, we focused on the effects of phthalates on -cell function (insulin secretion and storage) and -cell mass (flow cytometric cell cycle analysis). We selected phthalates as our target compounds because these ubiquitous compounds have been linked to an increased prevalence of insulin resistance. Both chronic and acute effects were evaluated and the underlying molecular pathways unraveled using microarray analysis. Our results show for the first time that phthalates are able to alter -cell function and therefore indicate that environmental pollutants might be involved in the development and progression of type 2 diabetes. doi:10.1016/j.toxlet.2011.05.220
OS11-3 In vitro carcinogenicity of perfluorooctane sulfonate (PFOS) on Syrian hamster embryo (SHE) cells N. Jacquet 1,∗ , M. Maire 2 , Y. Landkocz 2 , P. Vasseur 2 1
Université Paul Verlaine, Metz, France, 2 UMR CNRS 7146 – Université de Metz, Metz, France Perfluorooctane sulfonate (PFOS) (C8 F17 SO3 ) and its salts are persistent chemicals, bioaccumulative and toxic to mammalian species. Concerns have been raised about their widespread distribution on global earth, and their contamination of human serum. The Directive 2006/122/EC restricted the use of PFOS as substance or constituent of preparations. Yet, PFOS and their derivatives are still used in Europe for their hydrophobic and oleophobic properties as surface coating agents for textiles, photographic and electroplating processes, and also as foaming agents. PFOS has been reported to be a peroxisome proliferator. It is not mutagenic nor genotoxic, but is suspected to be carcinogenic. To elucidate this point, we evaluated carcinogenic potential of PFOS using the cell transformation assay on Syrian hamster embryo (SHE) cells, recommended by OECD (Organisation for Economic Co-operation and Development) for in vitro detection of chemical carcinogens. A range of PFOS concentrations from 2 × 10−5 to 50 g/ml (3 × 10−5 –100 M) was tested on SHE cells for neoplastic transformation and genotoxicity. A significant increase of cell transformation frequency was observed at non cytotoxic concentrations, equivalent to the ones found in human serum of PFOS-exposed workers. No genotoxicity was detected. Effects on PPARs gene expression were measured by qPCR and confirmed PPAR induction as one of PFOS modes of action. From the whole results, PFOS can be classified as a non-genotoxic carcinogen. An involvement of PFOS in increased incidence of cancer recorded in occupationally exposed population cannot be ruled out. doi:10.1016/j.toxlet.2011.05.221
The prevalence of diabetes mellitus is currently at epidemic proportions and it is estimated that it will increase even further over the next decades. Although genetic predisposition and lifestyle choices are commonly accepted reasons for the occurrence of type 2 diabetes, it has recently been suggested that environmental pollutants are additional risk factors for diabetes development. This project investigates if environmental pollutants might be involved in the development and progression of type 2 diabetes by focusing on effects on pancreatic -cells (both function and mass) and liver cells (insulin sensitivity) using in vitro models. However, as
S57
progresses. The LTKB-BD has good representation of known drugs in terms of chemical structure, therapeutic categories, and daily dosage. The LTKB-BD has the potential to serve as a benchmark dataset for the community-wide effort to evaluate the performance characteristics of existing DILI biomarkers and to develop novel DILI biomarkers using emerging molecular technologies. doi:10.1016/j.toxlet.2011.05.217
OS11 In Vitro Toxicology
OS11-1 Evaluation of drug-induced toxicity in isolated mouse liver mitochondria as a predictive model of hepatotoxicity risks in human M. Porceddu 1 , N. Buron 1 , C. Roussel 1 , B. Fromenty 2 , A. Borgne-Sanchez 1,∗ 1
MITOLOGICS SAS, Paris, France, 2 U991, INSERM, Rennes, France
Purpose: Mitochondrial dysfunction is a major mechanism of drug-induced liver injury (DILI), which can be severe and even fatal. DILI can thus lead to the interruption of clinical trials, or drug withdrawal after marketing. As the financial losses can be important for pharmaceutical companies, drug-induced mitochondrial dysfunction and DILI should be detected at early development stages. Methods: To facilitate this detection, we propose a high-throughput screening platform on isolated mouse liver mitochondria which allows the measurement of several parameters pertaining to mitochondrial function and membrane integrity. Indeed, these multiparametric assays measure: (1) global mitochondrial membrane permeabilization, (2) transmembrane potential, (3) mitochondrial outer membrane permeabilization and (4) respiration in isolated mouse liver mitochondria. Results: The screening of 100 drugs known to be hepatotoxic (Alpidem, Lumiracoxib, Troglitazone. . .) or not, allowed us to establish a strong correlation between in vitro mitochondrio-toxicity and DILI in human. A robust statistical analysis demonstrates the high predictivity level of this screening platform. doi:10.1016/j.toxlet.2011.05.219
OS11-2 Optimization of an in vitro model to study environmental pollutant effects on pancreatic -cell function and evaluation of the mode of action of phthalates T. Hectors ∗ , C. Vanparys, A. Pereira-Fernandes, R. Blust Ecophysiology, Biochemistry and Toxicology, University of Antwerp, Antwerp, Belgium
-cell failure is the triggering factor for the progression of prediabetic states to type 2 diabetes, there is a great need to focus on the impact of chemicals on -cells. Therefore, we investigated if the pancreatic -cell line INS-1 832/13 can be used as a system for the evaluation of compound effects and we optimized exposure conditions using pharmaceutical compounds. Secondly, we focused on the effects of phthalates on -cell function (insulin secretion and storage) and -cell mass (flow cytometric cell cycle analysis). We selected phthalates as our target compounds because these ubiquitous compounds have been linked to an increased prevalence of insulin resistance. Both chronic and acute effects were evaluated and the underlying molecular pathways unraveled using microarray analysis. Our results show for the first time that phthalates are able to alter -cell function and therefore indicate that environmental pollutants might be involved in the development and progression of type 2 diabetes. doi:10.1016/j.toxlet.2011.05.220
OS11-3 In vitro carcinogenicity of perfluorooctane sulfonate (PFOS) on Syrian hamster embryo (SHE) cells N. Jacquet 1,∗ , M. Maire 2 , Y. Landkocz 2 , P. Vasseur 2 1
Université Paul Verlaine, Metz, France, 2 UMR CNRS 7146 – Université de Metz, Metz, France Perfluorooctane sulfonate (PFOS) (C8 F17 SO3 ) and its salts are persistent chemicals, bioaccumulative and toxic to mammalian species. Concerns have been raised about their widespread distribution on global earth, and their contamination of human serum. The Directive 2006/122/EC restricted the use of PFOS as substance or constituent of preparations. Yet, PFOS and their derivatives are still used in Europe for their hydrophobic and oleophobic properties as surface coating agents for textiles, photographic and electroplating processes, and also as foaming agents. PFOS has been reported to be a peroxisome proliferator. It is not mutagenic nor genotoxic, but is suspected to be carcinogenic. To elucidate this point, we evaluated carcinogenic potential of PFOS using the cell transformation assay on Syrian hamster embryo (SHE) cells, recommended by OECD (Organisation for Economic Co-operation and Development) for in vitro detection of chemical carcinogens. A range of PFOS concentrations from 2 × 10−5 to 50 g/ml (3 × 10−5 –100 M) was tested on SHE cells for neoplastic transformation and genotoxicity. A significant increase of cell transformation frequency was observed at non cytotoxic concentrations, equivalent to the ones found in human serum of PFOS-exposed workers. No genotoxicity was detected. Effects on PPARs gene expression were measured by qPCR and confirmed PPAR induction as one of PFOS modes of action. From the whole results, PFOS can be classified as a non-genotoxic carcinogen. An involvement of PFOS in increased incidence of cancer recorded in occupationally exposed population cannot be ruled out. doi:10.1016/j.toxlet.2011.05.221
The prevalence of diabetes mellitus is currently at epidemic proportions and it is estimated that it will increase even further over the next decades. Although genetic predisposition and lifestyle choices are commonly accepted reasons for the occurrence of type 2 diabetes, it has recently been suggested that environmental pollutants are additional risk factors for diabetes development. This project investigates if environmental pollutants might be involved in the development and progression of type 2 diabetes by focusing on effects on pancreatic -cells (both function and mass) and liver cells (insulin sensitivity) using in vitro models. However, as
S57