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. This strategy can be applied to any type of chemical and systemic repeated dose toxicity effect. However, for proof-of-concept purposes, focus will be put on 6 specific NAMs addressing adversities in the liver (steatosis and cholestasis), kidneys (tubular necrosis and crystallopathy) and developing brain (neural tube closure and cognitive function defects) induced by a variety of chemicals, including from the pharmaceutical, cosmetics, food and biocide sectors. The 6 NAMs will each consist of a computational system based on cutting-edge artificial intelligence (AI) and will be primarily fed by available biological/mechanistic, toxicological/epidemiological, physico-chemical and kinetic data. Data will be consecutively integrated in physiological maps, quantitative adverse outcome pathway networks and ontology frameworks. Data gaps, as identified by AI, will be filled by targeted state-of-the-art in vitro and in silico testing. The 6 NAMs will be evaluated and applied in collaboration with industrial and regulatory stakeholders in order to maximise end-user acceptance and regulatory confidence. This is anticipated to expedite implementation in risk assessment practice and to facilitate commercialisation. ONTOX will have a deep and long-lasting impact at many levels, in particular by consolidating Europe’s world-leading position regarding the development, exploitation, regulation and application of animal-free methods for human risk assessment of chemicals.

The goal of PrecisionTox is to advance safety assessment of chemicals without the use of animal testing by establishing a new, 3Rs-compliant, cost-effective testing paradigm for chemical safety assessment — Precision Toxicology — that identifies molecular key event (KE) biomarkers predictive of chemically induced adverse health effects in humans and facilitates their uptake into regulatory and industry practice. This goal is supported by three core concepts: PhyloToxicology, which replaces mammal models with an evolutionarily diverse suite of non-sentient animal species from across the tree of life; Quantitative Susceptibility, which determines safety factors based on genetic variability; and Embedded Translation, which engages key stakeholders in project planning, selection of chemicals for investigation, and case studies for regulatory application. We accomplish this goal through six objectives: ● Stakeholder Integration, embedding the Stakeholder Advisory Group in project management (WP1); ● Comparative Toxicology, utilising high-throughput testing methods across five non-sentient species and human cell lines to observe toxic response (WP2); ● Molecular Data Production, applying metabolomics and transcriptomics to comparative toxicology samples to trace adverse outcomes via the molecular key events preceding them (WP3); ● Quantitative Susceptibility, applying quantitative genetics and gene expression profiling to understand variation in individual susceptibility and develop empirical exposure thresholds (WP4); ● Biomarker Discovery, PrecisionTox Data Commons, and NAM Toolbox, using machine learning to identify biomarkers for molecular key events and creating the dissemination and translation products for their use (WP5); and ● Regulatory Analysis and Application, partnering with JRC and regulatory agencies to identify opportunities for applying Precision Toxicology within existing regulatory structures and develop draft guidance for industry use and reporting (WP6).

The vision of the NAMWISE project is to provide a supporting framework to pave a scientifically substantiated way towards a paradigm-shift in (eco)toxicology that will not rely on in vivo data. To fulfil its vision the project will leverage the acquired knowledge on New Approach Methodologies (NAMs) and then gather new input from all stakeholders to refine, improve and optimize their use in regulatory contexts for chemicals and pharmaceuticals. The project will merge under its umbrella inputs from a multi-disciplinary consortium comprising academia, NAM-developers, NAM users (CROs), regulators while reaching out towards societal expectations. The workplan of NAMWISE is organized along far-reaching objectives driven by the interest for all the stakeholders: 1. Sustain and amplify the momentum built by similar initiatives by combining expertise on in silico tools, in vitro assays and a widespread network with external projects. 2. Provide a pragmatic approach based on case-studies for hazard/risk assessment and drug efficacy that will guide the project in implementing NAMs strategies. These procedures will be submitted to a peer-review to ensure that what the NAM community receive is of a high standard. 3. Deliver an analysis of requirements in terms of standardization and validation of NAMs that benefits from the input of the PEPPER platform on pre-validation, a unique French initiative on NAMs based on public and private funding. 4. Foster fruitful interactions between stakeholders catalyzed by several actions, including a series of workshops and training initiatives targeted on the needs of CROs identified during the project. 5. Propose a way forward for solving NAMs’ shortcomings and enhancing their drivers. Finally, the project will provide a white paper during its final open symposium that will render NAMWISE a major reference point and essential force for bringing forward proposals for animal-free chemical and drug assessments.

The toxicological impact of exposure to chemical mixtures is a matter of undisputed concern, but mixtures are only slowly making their way into regulatory risk assessment. Critical knowledge gaps are which and how many chemicals drive mixture effects in the environment and in humans. Scientific uncertainty remains on the validity of the dose addition principle for complex mixtures of large numbers of chemicals at low concentrations as they occur in our bodies. The PANORAMIX consortium addresses these challenges by showcasing a novel experimental path based on whole mixture assessments for identifying and quantifying the risk of chemical mixtures extracted from real-life samples representing environment and food as well as humans. We provide ready-to-use and practical tools for mixture risk assessment of several chemicals with a diverse range of adverse health outcomes. The applied methodologies, including a panel of in vitro assays coupled with effect-directed analyses and large-scale suspect and non-targeted chemical profiling are innovative in their combinatorial approach. Specifically, we will take advantage of a well-studied human cohort of new-borns, in whom adverse health outcomes related to developmental toxicity originating from chemical mixture exposure will be identified. PANORAMIX will use mixture modelling, case studies and experimental data to deliver a web-based interface for calculating risks to chemical mixtures and to define effect-based trigger values for in vitro effects that can be directly measured in water, food, and blood to identify when mixture exposure is posing a health threat. By involving regulatory and scientific stakeholders throughout the project, we support the implementation of existing mixture risk assessment and management approaches to reduce the most critical exposures and assist in optimizing regulatory approaches to yield evidence-based policies, contributing to EU's zero-pollution ambition for a toxic free environment in the future.