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JRC and OECD activities related to EuroMix

Andrew Worth & Stephanie Bopp

European Commission Joint Research Centre

Institute for Health & Consumer Protection

Systems Toxicology Unit

DISCLAIMER: This presentation and its contents do not constitute anofficial position of the European Commission or any of its services.Neither the European Commission nor any person acting on behalf ofthe Commission is responsible for the use which might be made ofthis presentation or its contents

EuroMix Kick-off meeting, RIVM, Bilthoven, 20 May 2015

Overview

• Introduction to the JRC and EURL ECVAM

• Dissemination of information on alternative methods

• Predictive toxicology research within Seurat-1

• Adverse Outcome Pathway (AOP) development activities

• JRC and OECD activities on the assessment of chemical mixtures

• Concluding remarks

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Institute for Health and Consumer Protection

Institute for Prospective Technological Studies

Institute for Reference Materials and Measurements

Institute for Transuranium Elements

Institute for Energy and Transport

Institute for the Protection and Security of the Citizen Institute for Environment and Sustainability

Ispra Site Management

Headquarters

The Joint Research Centre (JRC) -the European Commission’s in-house science service

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~ 70 staffGroups Assay

ValidationPredictive Toxicology

Toxicity Pathways

HTSHCS

In Vitro GLP

Facilities

Endocrine disruptors

Combined exposures and chemical mixtures

Standardisation and international harmonisation of alternative methods

Information systems for safety assessment and alternative methods

Integrated approaches to testing and assessment

Projects: http://projects.jrc.ec.europa.eu

Systems Toxicology Unit / EURL ECVAM

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Regulation 1907/2006 on registration, evaluation, authorisation and restriction 

of  chemical substances (REACH)

Regulation 440/2008 on test methods

Regulation 1272/2008 on classification, labelling and packaging of chemical 

substances and mixtures (CLP)

Regulation  1223/2009 on cosmetics products

Regulation 528/2012 on authorisation of biocidal active 

ingredients an products

Community Strategy on Endocrine Disrupters

Directive 91/414 on authorisation of plant protection active 

ingredient  and products

Community Strategy on combined  

exposures 'Mixtures'

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Support to chemicals policy

o Coordinate and promote development and use

o Coordinate validation at Union level

o Information exchange on development

o Databases and information systems

o Promote dialogue between legislators, regulators and stakeholders

Key responsibilities*

Established under Directive 2010/63/EU on the protection of animals used for scientific purposes

*Article 48 of the Directive, Annex VII

The European Union Reference Laboratory for Alternatives to Animal Testing

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• Analysis of regulatory information

needs across sectors

• Broad strategic aims to promote

solutions for regulatory acceptance

Available

• Skin sensitisation

• Genotoxicity

• Aquatic toxicity

• Acute systemic toxicity

https://eurl-ecvam.jrc.ec.europa.eu/eurl-ecvam-strategy-papers

EURL ECVAM Strategy Documents

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In preparation

• Toxicokinetics

• (Developmental) Neurotoxicity

https://eurl-ecvam.jrc.ec.europa.eu/eurl-ecvam-2014-report-alternative-methods

EURL ECVAM Status Reports

https://eurl-ecvam.jrc.ec.europa.eu/news/review-of-available-methods

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DB-ALM: EURL ECVAM DataBase service on ALternative Methods

http://ecvam-dbalm.jrc.ec.europa.eu Contact: Annett Janusch-Roi9

JRC QSAR Model Database

http://qsardb.jrc.ec.europa.eu Contact: Jutta Triebe10

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The SEURAT strategy is to adopt a toxicological mode-of-action framework todescribe how any substance may adversely affect human health, and to use thisknowledge to develop complementary theoretical, computational andexperimental (in vitro) models that predict quantitative points of departureneeded for safety assessment.

• Cluster of seven collaborative projects• 50 million Euro investment• Co‐financed by EC and Cosmetics Europe• Over 70 research partners• 16 countries plus EC• 6 year programme

Seurat-1: towards replacement of in vivo repeated dose systemic toxicity testing

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COSMOS

Integrated In Silico Models for the Prediction of Human Repeated Dose Toxicity of Cosmetics to Optimise Safety

• Collection of toxicological data

• Development of the Threshold of Toxicological Concern (TTC) approach

• Development of novel in silico methods

• Multiscale modelling:

mitochondrial (dys)function, virtual cell-based assay, 2D liver,Physiologically Based Biokinetic (PBBK) models

• In silico workflows based on open-source and open-access tools

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COSMOS database v1.0

• Open-access

• High-quality toxicity data (quality control, structure curation)

• User-friendly query builder (chemical name, structure, toxicity data)

• 44,765 unique chemical structures

• 12,538 toxicity studies for 1,660 compounds across 27 endpoints

Webinar and tutorial:

http://www.cosmostox.eu/what/COSMOSdb/

http://cosmosdb.cosmostox.eu/

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External Dose

HTS - In vitro concentration

Multi-scale modelling

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ExposureMolecular Initiating

Event

Organelle Effects

Cellular Effects

Tissue Effects

Organ Response

Individual Response

Population Response

Toxicity Pathway

Adverse Outcome Pathway (human health effect)

Adverse Outcome Pathway (environmental effect)

Adverse Outcome Pathway approach

Molecular Initiating Event

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Responsibility of the Extended Advisory Group on Molecular Screening and Toxicogenomics (EAGMST)

Co-chairs: Robert Kavlock (US EPA) & Maurice Whelan (EC JRC)

OECD AOP Development Programme

• Development of an AOP

(applicable to a chemical category)

• Development of an AOP Case Study

(applicable to a single or few chemicals)

• Guidance Document (AOP development including evaluation)

• Knowledge Management Tools

Project categories:

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AOP Knowledge Base

Qualita

tive

calibration

http://aopkb.org/aopwiki 17

AOPs related to developmental & reproductive toxicity

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• Androgen receptor agonism leading to reproductive dysfunction

• Aromatase inhibition leading to reproductive dysfunction (in fish)

• Estrogen receptor antagonism leading to reproductive dysfunction

• Xenobiotic induced inhibition of thyroperoxidase and subsequent adverse

neurodevelopmental outcomes in mammals

• PPARα activation leading to impaired fertility in male rodents (JRC)

• PPARγ activation leading to impaired fertility in adult female rodents (JRC)

• Inhibition of mitochondrial complex I leading to neurodegeneration (JRC)

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AOPs related to liver toxicity

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• Protein alkylation leading to liver fibrosis (JRC)

• LXR activation to liver steatosis (JRC)

• Cholestatic liver injury induced by inhibition of the bile salt export pump

• NFE2L2/FXR activation leading to hepatic steatosis

Whole mixture testing vs component-based testing and modelling approach

Scientific approaches

Dissimilarly acting substances: Independent ActionMultiply constitutent effects

Similarly acting substances: Concentration AdditionAdd equipotent doses

InteractionsTotal effect greater or less than additive effect

Use of AOP network?

Nodes represent Key Events (KE)Edges represent Key Event Relationships (KER) 20

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JRC activities on the assessment of chemical mixtures / combined

exposures

Some definitions

Intentional mixtures: manufactured products or formulations, including commercial mixtures of industrial substances (e.g. pesticide formulations)

Unintentional mixtures: substances from different sources, deposited separately at a particular site (e.g. in surface water)

Aggregate exposure: exposure to single substance originating from different sources (e.g. used as pesticide and as food/feed additive)

Combined exposure: multiple substances from one or different sources

Chemical X Chemical YChemical XChemical X Chemical Z

Chemical n Chemical …

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Regulatory RA approach mainly focuses on the assessment of individual substances, potential gap in RA framework identified:

• Kortenkamp 2009 State of the Art Report on mixture toxicity

• SCHER/SCENIHR/SCCS 2011 Opinion on toxicity assessment of chemical mixtures

• Commission Communication on the combined effects of chemicals (COM(2012)252 final)

Past EC activities

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Past EC activitiesCurrent situation

"…no mechanism for a systematic, comprehensive and integrated assessment of mixture effects taking into account different routes of exposure and different product types."

For human tox: safe levels established for individual chemicals are considered sufficiently protective for mixtures of dissimilarly acting compounds. For chemicals with similar MoA potential for cumulative effects; Concentration/dose addition approach preferred.

For ecotox: possible combination effects for both similar and independently acting chemicals due to protection goal at population level

Follow up actions:

Coherent approach for generation, collection storage and use of chemical monitoring data (IPChem)

Address knowledge gaps with regard to MoA of chemicals, grouping of chemicals, predicting interactions, identify substances that are drivers of mixture toxicity (supported by e.g. H2020 activities)

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Regulatory Requirements and GuidanceReviewed EU legislations • Intentional: CLP REACH PPPR BPR

• Unintentional/environmental: Food contaminants Food contact materials Waste streams WFD Marine Stategy Groundwater Drinking Water

medicines cosmetics food /feed related

PPP MRL Soil Air EIA IPPC Toys

JRC Review

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Regulatory Requirements and Guidance

JRC Review

Reviewed Guidance• European Guidance Documents

for (eco)toxicity assessment of mixtures (e.g. ECHA, EFSA,…)

• US EPA, Canada, other national proposals

Internationally proposed frameworks for assessing chemical mixtures:• WHO/IPCS 2009• SCHER, SCENIHR, SCCS 2011 • CEFIC MIAT 2012

no harmonised approach or clear guidance

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JRC survey Respondents

• Aim: collect information on approaches, experiences and future directions in assessing human and environmental health risks from chemical mixtures

• Approach: online survey Jan-Mar 2015, send to experts in the field selected from relevant publications and workshops

Respondents:• 58 replies• from 16 EU and 5 other

countries• 48 have performed mixture

assessments

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JRC survey Experiences

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Authorisation of commercial products(i.e. prospective RA)

Research and development

General exposure monitoring

RA of contaminated sites

Priority setting of risk reductionmeasures

Control of emission permits

Control of remediation works and theirsuccess

Other

For which purpose(s) did you assess the overall toxicity of chemical mixtures?

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JRC survey ExperiencesHave you ever applied whole mixture (WM) or component based (CB) approaches for the assessment of intentional (int.) or unintentional (unint.) mixtures?

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JRC survey Expert opinions

Which type of mixture(s) or samples would you identify as highest priority for RA that needs to take mixture effects into account?

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Based on your experience or knowledge about the toxicity of mixtures, do you consider it important to include interactions (e.g. potential synergism or antagonism) in the assessment?

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JRC survey Expert opinions

Do you apply novel tools in the RA of mixtures?

No reply

No

Yes for HRA+ERA

Yes for ERA

Yes for HRA

• Many experts are already using such novel tools

• Up to now mainly considered research activity, but start being applied also in regulatory context

Most often mentioned reasons for notapplying such tools are: • Lack of legal drivers• Lack of standardisation/validation• Lack of guidance• Lack of expertise 32

JRC survey Expert opinions

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OECD Working Group on Combined Exposure

Aims:

• Exchange knowledge and experience related to the assessment of risk from the combined exposure to multiple chemicals

• Illustrate general considerations drawing from case studies

• Move towards technical convergence on scientific considerations between member countries

• Build upon the recommendations from the WHO OECD ILSI/HESI International Workshop on Risk Assessment of Combined Exposures to Multiple Chemicals

JRC activities linked to OECD

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OECD Working Group on Combined Exposure

Areas covered:

1. Problem formulation regarding prioritization/triggers/scope for assessment of combined exposures

2. Considerations for hazard characterization to inform assessment of combined exposures

3. Considerations regarding co-exposure characterization to inform assessment of combined exposures

4. Considerations regarding risk assessment of combined exposures using various approaches and capturing and communicating uncertainties in findings

JRC activities linked to OECD

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Considerations for hazard assessment

1. How to combine chemicals into hazard (sub)categories based on expected adverse outcome and/or MoA

2. Considerations for incorporation of different data types (e.g. in silico, in vitro, in vivo methods)

3. How to consider differences in potency of chemicals

4. Consider data needs for moving through tiers of hazard assessment within WHO/IPCS framework (data types, uncertainties)

JRC activities linked to OECD

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• In order to do the RA, chemicals within a mixture can be grouped according to common effects (not only similar structure/phys-chem properties but effect data/MoA)

• AOPs can help to identify common/similar effects for grouping, identify data gaps and strategies to fill them

How AOPs could help in grouping of chemicals

JRC activities linked to OECD

Common target organsRefine using phenomenological effects

Common Molecular Initiating Event (MIE) and early key events

Use all available information on common events from all levels 36

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Conclusions

• Further research and tools are needed for a harmonised assessment of chemical mixtures/combined exposure

• EuroMix can make a substantial contribution to this need addressing various aspects of what is currently under debate

• EuroMix should ensure to keep informed relevant stakeholders (both directions)

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Acknowledgements

Mixture team in JRC:

Aude KienzlerSander van der LindenJos BessemsElisabet Berggren

Contact:andrew.worth@ec.europa.eu

stephanie.bopp@ec.europa.eu