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DSA | Laboratoire de Spectrométrie de Masse BioOrganique (LSMBO) » NOTOX project

Predicting long-term toxic effects using computer models based on systems characterization of organotypic cultures

Dernière mise à jour : jeudi 18 juillet 2013, par Nicolas Busser

The EU ban on the sale of cosmetics whose ingredients have been tested on animals entered into force on March 11, 2013. However, before being launched on the European market, new cosmetic and hygiene products need to pass a thorough safety assessment to ensure their ingredients have no detrimental effects on human health. There is, therefore, an urgent need for alternative methods. The replacement of in vivo testing for systemic, repeated dose and long term toxicity in humans still represents a major challenge. The existing long term test methods involving animals can only be replaced by incorporating various strategies in an integrated multifaceted platform using a systems biology approach. This also require in silico methods and powerful bioinformatic tools to process huge amounts of data that is usually collected in a typical systems biological setup. The European NOTOX project (, as part of the European research initiative SEURAT-1 (, develops systems biology approaches for in vitro toxicity studies and in vitro-in vivo extrapolation.

The NOTOX project : one of the building block projects of the European research initiative SEURAT [1] co-funded by the European Commission [2] and the European trade association Cosmetics Europe [3].

Contact : Fabrice BERTILE

NOTOX main goals

Predicting long-term toxic effects using computer models based on systems characterization of organotypic cultures.

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The overall objectives of NOTOX are to identify cellular and molecular signatures of long term hepatotoxicity and to validate predictive mathematical and bioinformatic models characterizing long term toxicity responses. To this end, NOTOX uses the human HepaRG cell line as well as primary human liver cells and develops a spectrum of systems biological tools including experimental and computational methods for organotypic human cell cultures suitable for long term toxicity testing and the identification and analysis of pathways of toxicological relevance.

Cellular activities are monitored continuously by comprehensive analysis of released metabolites, peptides and proteins and by the estimation of metabolic fluxes using 13C labelling techniques (fluxomics). At selected time points a part of the cells are removed for in-depth structural (3D-optical and electron microscopy tomography), transcriptomic, epigenomic, metabolomic, proteomic and fluxomic characterizations. Physiological data including metabolism of test compounds will be incorporated into large-scale computer models that are based on material balancing and kinetics. Various “-omics” data and 3D structural information from organotypic cultures will be integrated using correlative bioinformatic tools. These data also serve as a basis for large scale mathematical models.

This systems approach is highly promising for a more comprehensive understanding and better prediction of repeated dose toxicity of test compounds.

[ Learn more on NOTOX ]

NOTOX film – take a look behind the scenes !

IPHC-LSMBO contribution for NOTOX

The current “gold standard” for in vitro toxicological studies, primary human hepatocytes (PHH), suffers from inherent restrictions like limited and unpredictable availability, pronounced donor-to-donor variances as well as rapid loss of liver specific functions under in vitro conditions. Recently, the hepatic cell line HepaRG (Biopredic International) has gained much attention as a human hepatocyte model system suitable for long-term cultivation up to several weeks. Although several studies investigated the transcriptome of HepaRG cells, proteomic data remains scarce.

Therefore, the LSMBO (Laboratoire de Spectrométrie de Masse Bio-Organique) is in charge of the proteomics characterization of HepaRG cells, and of identification of acute and chronic drug-induced hepatotoxic effects at the protein level.

We develop complementary proteomics analytical strategies to identify and quantify proteins :

  • two-dimensional difference gel electrophoresis (2D DIGE) coupled to mass spectrometry (MS), which allows the high-throughput discovery of high abundance proteins that are differentially regulated in response to a given treatment.
  • label free quantitative shotgun proteomics analysis, which allows the high-throughput discovery of low to high abundance proteins that are differentially regulated in response to a given treatment.
  • selected reaction monitoring (SRM)-mass spectrometry (MS), which allows hypothesis-driven quantitation of a restricted number of targeted proteins.

Main IPHC members working on the NOTOX project :

Dr. Alain Van Dorsselaer (Coordinator of proteomics studies)
Dr. Fabrice Bertile (Scientific responsible for proteomics studies)
Dr. Georg Tascher (postdoctoral researcher)
Miss Marine Plumel (PhD student)

Project Coordination :

Prof Dr. Elmar HEINZLE
Biochemical Engineering Institute
Saarland University
Tel : +49 - 681- 302 2905 or 3405
Fax : +49 - 681- 302 4572

NOTOX consortium

NOTOX has assembled top academic experts and SMEs from France, Sweden, The Netherlands, Israel, Great Britain and Germany. The project is coordinated by Prof. Elmar Heinzle at the Biochemical Engineering Institute of the Saarland University together with Dr. Fozia Noor and EURICE.

Partners :

  • Universität des Saarlandes, Germany (Prof E Heinzle, Dr. F Noor, Prof. J Walter)
  • CNRS, IPHC, France (Dr. A van Dorsselaer, Dr. F Bertile)
  • NKI-AVL, Stichting Het Nederlands Kanker Instituut, The Netherlands (Prof PJ Peters)
  • Karolinska Institutet, Sweden (Prof M Ingelman-Sundberg)
  • Insilico Biotechnology, Germany (Dr. K Mauch)
  • INRIA, Institut National de Recherche en Informatique et en Automatique, France (Dr. D Drasdo)
  • DFKI, Deutsches Forschungszentrum für Künstliche Intelligenz, Germany (Prof P Slusallek)
  • IFADO, Leibniz Research Centre for Working Environment and Human Factors, Germany (Prof J Hengstler)
  • Biopredic International, France (Dr. C Chesné)
  • Weizmann Institute of Science, Israel (Dr. A Tanay)
  • Cambridge Cell Networks, United Kingdom (Dr. G Apic)
  • EURICE, European Research and Project Office, Germany (C Schacht, Dr. V Peuser)

About SEURAT-1

SEURAT-1 plays an important role in continuing to find the alternative methods replacing animal testing. This initiative is composed of seven research projects, which closely cooperate with a common goal and combine the research efforts of over 70 European universities, public research institutes and companies. The projects were selected through the call for proposals “FP7-HEALTH-2010-Alternative-Testing-Strategies”. They received funding support from the European Commission’s FP7 HEALTH Programme and from Cosmetics Europe. SEURAT-1 started on 1 January 2011 and will run for five years.

The Research Initiative is a first step to addressing the long term strategic target of "Safety Evaluation Ultimately Replacing Animal Testing (SEURAT)". It is called "SEURAT-1", indicating that more steps have to be taken before the final goal will be reached. SEURAT-1 develops knowledge and technology building blocks required for the development of solutions for the replacement of current repeated dose systemic toxicity testing in animals used for the assessment of human safety.

SEURAT-1 will contribute to develop a new definition of "adversity" in toxicology at the cellular and molecular level leading to pathway-based human safety assessment. This will require the establishment of a complex system consisting of stable human cell lines and organ-simulating devices in combination with computational chemistry, systems biology and sophisticated modeling and estimation techniques. Emphasis will be put on the reliability and reproducibility of all components necessary for the implementation of the concept.

SEURAT-1 will provide an essential step towards the SEURAT long term strategy by providing the proof of concept that the new toxicity pathway strategy reliably predicts risk of repeated dose human (liver) toxicity

[ Learn more on SEURAT-1 ]

[1SEURAT : safety evaluation ultimately replacing animal testing -

[2European Commission : 7th Framework Programme -

[3Cosmetics Europe - The Personal Care Association -