The vision of the ONTOX consortium is to provide a functional and sustainable solution for advancing human risk assessment of chemicals without the use of animals in line with the principles of 21st century toxicity testing and next generation risk assessment. Specifically, ONTOX will deliver a generic strategy to create innovative new approach methodologies (NAMs) in order to predict systemic repeated dose toxicity effects that, upon combination with tailored exposure assessment, will enable human risk assessment.
The Validation Management Team for the Multi-ImmunoTox Assay (MITA), with 3RsMC as member, published the outcome of an international validation study of this in vitro method evaluating the effects of immunotoxic chemicals on IL-2, IFN-γ, IL-1β and IL-8 promoter activities.
Twelve challenging substances were assessed in 4 reconstructed human epidermis (RHE) models. For hazard identification, the overall predictive accuracy ranged around 70% for three assays, although for one it fell below 50% when human data was used as the benchmark. These results indicate that the 3D RHE models may be a useful tool for assessing skin sensitisation potentials without needing to revert to animal use
The focus of this workshop was on recent advancements in our knowledge and understanding of respiratory allergy and including the development of an adverse outcome pathway (AOP) for respiratory sensitizers under auspices of the OECD (Sullivan et al., 2017, Applied In Vitro Toxicology). The workshop speakers described current and promising in silico/in chemico/in vitro/in vivo methodologies that may be used to develop a testing strategy for respiratory sensitizers. Furthermore, both the regulatory needs and industry perspective were addressed.
This workshop provided a forum for dialogue between the scientific, industry and regulatory communities which may foster a better understanding of key public health and occupational safety concerns and the best approaches to identify and protect against the adverse health outcomes of respiratory sensitizers
The application of the #AOP concept to structure mechanistic in vivo and in vitro knowledge has made it possible to identify a number of methods, each addressing a specific KE, that provide information about the #food allergenic potential of new #proteins.
When applied in the context of an integrated strategy these methods may reduce, if not replace, current animal testing approaches.
The proposed AOP for food sensitization induction will be shared at the www.aopwiki.org platform to expand the mechanistic data, improve the confidence in each of the proposed KE and key event relations (KERs), and allow for the identification of new, or refinement of established KE and KERs. (van Bilzen et al., 2017, Clinical and Translational Allergy)
In the context of this Stakeholder Meeting #3RsMC contributed to the first steps towards the creation of a platform for analysis of novel food and food proteins based on test methods and strategies proposed by the impARAS WG2. The focus was on technology and knowledge gaps, reliability, relevance and industrial applicability. The discussion was organized in 3 separate focus areas: 1. Prioritization of proteins for testing (In silico); 2. Test methods addressing innate responses covering the identified MIEs and KE; 3. Test methods addressing adaptive responses covering identified KE. 2. It was agreed to estblish an overview of the cell models available and their competence for testing control proteins which are currently available within the impARAS consortium. It was agreed to publish this information in a peer-reviewed manuscript.
The European Commission organized the scientific conference ‘Non-animal approaches: The way forward’ (#NonAnimalScience) in response to the European Citizens’ Stop Vivisection initiative. The conference succeeded in getting the relevant stakeholders in open dialogue on how to exploit advances in science towards the goal of phasing out animal testing.
I am looking forward to the publication of the conclusions and recommendations from the individual sessions, and hopefully perspectives on the way forward to #animalfreetesting. These recommendations should include the need for ‘An Overall European Strategy’, and the need for harmonization with e.g. the already ongoing US initiative. A top-to-bottom strategy is to bring scientific, technological as well as regulatory innovation together into one meltingpot from which useful methods for animalfree testing and assessment of human and environmental health and safety can be forged. The nature and format of these methods should be defined by the needs of the regulatory authorities, as well as by their applicability in an industrial context.
We may also need to innovate in terms of communication, e.g.communication as to why animal models, inspite of their limitations, may be required to make the transition become reality without jeopardizing human and environmental health and safety.
Efforts must be made to properly address the concerns raised by the European Citizens’ Stop Vivisection initiative. While the first session of the Conference was clearly questioning the added value of animal models, the messages from the other sessions were addressing the initiative in a more indirect, if not cryptic, manner and may not be understood. One conclusion of the conference was that our communication is not good enough. We could start here with improving.
3RsMC ApS became member of the In Vitro Testing Industrial Platform (IVTIP).
Visit at www.ivtip.org
In vitro Testing Industrial Platform (IVTIP) and the Centre for Alternatives to Animal Testing (CAAT) co-organized an event, hosted by the Liberals and Democrats for Europe Party, bringing together SMEs from the biotech sector and Members of the European Parliament (MEPs). The event was titled “Meet Your Biotech MEPs at the European Parliament” and took place in Brussels, where it gathered approximately 40 participants including SME representatives, MEPs, public affairs professionals and a representative of the European Partnership for Alternative Approaches to Animal Testing (EPAA).
Presentations were given by representatives of EASME (the Executive Agency of the European Commission in charge of SMEs), CAAT, IVTIP, Innovative Medicine Initiative (IMI) 2, and DG GROW (COSME programme for SMEs). 3RsMC ApS presented for IVTIP.
The participating MEPs included Philippe de Backer, Michèle Rivasi, Keith Taylor, Jeppe Kofod, Anja Hazekamp, Stephan Eck and Frederick Federley who hosted the whole event at the Parliament. They were very supportive of this initiative. They voiced their support to SMEs as a promising pathway to promote alternative methods and their validation.
This event was a follow-up of a CAAT workshop that took place earlier this year that linked MEPs with experts on alternatives.
The work of a dedicated consortium
3Rs Management and Consulting, Adriaens Consulting, Oroxcell, Yves Rocher, Amorepacific, University of Milan, MB Research Laboratories, CellSystems, Pierre-Fabre, Vrije Universiteit Medical Center, MatTek Cooperation, Eurofins, MatTek IVSL, Stiefel
Conclusion
This study has demonstrated that the RHE IL-18 potency test for skin sensitization is easily transferred from the test developer to a naive laboratory. It also identified critical issues that need to be controlled if quantitative data have to be compared (e.g. for relative potency determination). The consortium is currently preparing an extensive validation of the updated protocol. The planned validation study compares the three tissue models for reliability, predictive capacity and limitations in terms of chemical applicability domain.
Results
The training and technology transfer of the protocol were successful as the participants in this study produced the same prediction as the test developers providing the training. At subtoxic levels the SkinEthic™ RHE model produced basal IL-18 levels that were consistently >10x higher than the basal levels observed with the epiCS® and MatTek EpiDerm™ models. This did not significantly affect stimulation in IL-18 levels triggered by the vehicle controls (< 2x). Considerable variation (< 5x) in cell viability and IL-18 stimulation was observed after exposure to sensitizers. These variations correlate with the RHE models and the ELISA used to determine IL-18 levels. Statistical analysis of the data obtained for the five test substances tested under the different experimental conditions has resulted in an acceptance criteria (90% Confidence Interval) check list that is incorporated in the updated protocol of the RHE IL-18 potency test for skin sensitization.
Continues Education Course, Sunday September 13, 10:00-12:00.
The presentation will focus on the importance of the ‘undescribed’ for industrial and regulatory application of animal-free methods for safety assessment.
1. The requirements to apply animal-free methods do not hit all industry sectors ans companies equally. This has unintended consequences for the competitiveness of especially Small and Medium Enterprises (SMEs).
2. There are concerns about compliance with the requirement to prioritize animal-free methods.
- Recognizing Test Guidelines only blocks the application of informative, scientifically validated, mechanism-based methods.
- Industry has to deal with a common safety evaluation strategy caught in a net of vertical (e.g. industry specific legislation) and horizontal (e.g. EU legislation 2010/63/EU).
3. There is too little incentive to start working with the available (not necessarily validated) methods and to create high quality data that can be used to convince regulatory authorities. Innovation and pragmatism will have to be combined to make this possible.
Background:
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