Periodic Reporting for period 2 - RISK-HUNT3R (RISK assessment of chemicals integrating HUman centric Next generation Testing strategies promoting the 3Rs)
Período documentado: 2022-12-01 hasta 2024-05-31
Science and society have increasingly started demanding a paradigm shift towards chemical risk assessment without using animals. In addition to growing ethical concerns around the use of animals, there is a fundamental need to address all potential adverse effects on human health. Animals may not be the best model to predict all human adversities. Furthermore, the limited throughput of animal-based testing in European laboratories (high costs, insufficient laboratory capacity) currently prevents the comprehensive evaluation of all chemicals that require safety assessment.
Scientific advances in in silico and in vitro and in vivo testing, via so-called new approach methodologies (NAMs), have greatly increased the opportunities for application of NAMs in chemical safety testing.
The RISK-HUNT3R (RH3R) vision is that animal-free next generation risk assessment (NGRA) can be implemented through the combination of human exposure assessment and NAM-based toxicokinetic and toxicodynamic testing/modelling. These will be integrated in a risk assessment framework using computational approaches and final decisions based on weight of evidence. To achieve this, the project hosts leading experts from many disciplines aiming to develop a novel and modular framework for NGRA. Furthermore, RH3R closely works with regulators including European regulatory agencies to enable the translation of research findings to risk assessment practice.
The aim of RH3R is to better protect citizens against hazardous chemicals by the development and use of non-animal testing approaches. This is in line with the goal of Europe’s Green Deal and with the need to provide an expertise and testing hub for European industries. RH3R will help achieve this by showing how NAMs can protect the human population, not just from acute adversities, but also from multifactorial diseases and chronic effects.
Scientific advances in in silico and in vitro and in vivo testing, via so-called new approach methodologies (NAMs), have greatly increased the opportunities for application of NAMs in chemical safety testing.
The RISK-HUNT3R (RH3R) vision is that animal-free next generation risk assessment (NGRA) can be implemented through the combination of human exposure assessment and NAM-based toxicokinetic and toxicodynamic testing/modelling. These will be integrated in a risk assessment framework using computational approaches and final decisions based on weight of evidence. To achieve this, the project hosts leading experts from many disciplines aiming to develop a novel and modular framework for NGRA. Furthermore, RH3R closely works with regulators including European regulatory agencies to enable the translation of research findings to risk assessment practice.
The aim of RH3R is to better protect citizens against hazardous chemicals by the development and use of non-animal testing approaches. This is in line with the goal of Europe’s Green Deal and with the need to provide an expertise and testing hub for European industries. RH3R will help achieve this by showing how NAMs can protect the human population, not just from acute adversities, but also from multifactorial diseases and chronic effects.
From the start of the project, we have been cementing our three main pillars: setting up our science and technology, making sure that our solutions can work in a regulatory context, and recruiting and engaging the people that share our vision and make it a reality.
Ultimately, we want to be able to answer the question whether a certain compound is safe, by using a safety testing pipeline that works for humans and that does not rely on animal testing. To this end, we are developing our NGRA workflow (ASPA) that is modular in nature, meaning that it is flexible and future-proof. Synergy on these activities has been achieved by common working groups within the joint ASPIS cluster.
Where the first project period saw the setting up and testing of the various workflow modules in exploratory studies, the second period focused more on their integration. To this end over 10 case studies has been delineated. Data generation and interpretation for these cases is greatly advancing and is guided by the developing ASPA in a tiered fashion.
To test if a chemical will enter the human body upon exposure, we implemented in vitro tests for uptake via the lung or gut. Moreover, our biokinetic models can predict concentrations in our tests while accounting for metabolism in the ADME pillar of ASPA, as well as for biological variability.
To predict toxicological hazards that might occur when chemicals are taken up by the body, we established computational pipelines based on chemical structures of compounds to predict to which targets they may bind for various tissue types. We also generated a panel of pluripotent stem cells with stress response reporters for high-throughput hazard screening in different target organ lineages, like hepatocytes, renal proximal tubular cells and cardiomyocytes. High-throughput data for over 100 compounds include phenotypic, MIE (molecular initiating event) and KE (key event) assay data.
For transcriptomics, which can show changes in all cellular processes at the same time, we set up comprehensive toxicogenomics prediction platforms for kidney and liver cells, including stem cell-based models for these cell types, as well as for mature peripheral neurons. We also set up state-of-the-art combined transcriptome and metabolome studies to determine cell fate. Kidney and liver organoids were further developed, used for transcriptome mapping, and combined in a two-organ chip. Challenging compounds were assembled and tested to show how to avoid false negatives.
Using network mapping, we delineated gene and protein networks and compounds associated with kidney toxicity, as well as putative new kidney biomarkers. We can connect transcriptomic and morphological data with clinical information and we constructed a database for all project compounds based on existing human exposome knowledge. We can quantify AOPs, all the way from MIE to late KEs, and developed a framework to evaluate such qAOPs. Meanwhile we started to implement our uncertainty framework for application in NGRA.
RH3R put in place a data and knowledge infrastructure with a harmonised data template and compound database, always respecting FAIR data storage criteria. For our NAMs we keep track of readiness levels for relevant applications, required for their success and commercial prospects. We also refined the ASPA workflow via interaction with stakeholders (regulators, industry, academia, NGOs) at multiple international events. A major innovation that has started to bring ASPA to the end-users is the NAMASTOX online dashboard to transparently operate on the workflow whilst integrating and documenting the safety assessment procedure.
RH3R established a strong internet presence. Key project publications and events can be found at the project website, in newsletters, and social media.
Lastly, RH3R has continuously provided highly rated training on NGRA to the new generation of scientists on risk assessment, including the practical context of linking lab research and regulatory reality.
Ultimately, we want to be able to answer the question whether a certain compound is safe, by using a safety testing pipeline that works for humans and that does not rely on animal testing. To this end, we are developing our NGRA workflow (ASPA) that is modular in nature, meaning that it is flexible and future-proof. Synergy on these activities has been achieved by common working groups within the joint ASPIS cluster.
Where the first project period saw the setting up and testing of the various workflow modules in exploratory studies, the second period focused more on their integration. To this end over 10 case studies has been delineated. Data generation and interpretation for these cases is greatly advancing and is guided by the developing ASPA in a tiered fashion.
To test if a chemical will enter the human body upon exposure, we implemented in vitro tests for uptake via the lung or gut. Moreover, our biokinetic models can predict concentrations in our tests while accounting for metabolism in the ADME pillar of ASPA, as well as for biological variability.
To predict toxicological hazards that might occur when chemicals are taken up by the body, we established computational pipelines based on chemical structures of compounds to predict to which targets they may bind for various tissue types. We also generated a panel of pluripotent stem cells with stress response reporters for high-throughput hazard screening in different target organ lineages, like hepatocytes, renal proximal tubular cells and cardiomyocytes. High-throughput data for over 100 compounds include phenotypic, MIE (molecular initiating event) and KE (key event) assay data.
For transcriptomics, which can show changes in all cellular processes at the same time, we set up comprehensive toxicogenomics prediction platforms for kidney and liver cells, including stem cell-based models for these cell types, as well as for mature peripheral neurons. We also set up state-of-the-art combined transcriptome and metabolome studies to determine cell fate. Kidney and liver organoids were further developed, used for transcriptome mapping, and combined in a two-organ chip. Challenging compounds were assembled and tested to show how to avoid false negatives.
Using network mapping, we delineated gene and protein networks and compounds associated with kidney toxicity, as well as putative new kidney biomarkers. We can connect transcriptomic and morphological data with clinical information and we constructed a database for all project compounds based on existing human exposome knowledge. We can quantify AOPs, all the way from MIE to late KEs, and developed a framework to evaluate such qAOPs. Meanwhile we started to implement our uncertainty framework for application in NGRA.
RH3R put in place a data and knowledge infrastructure with a harmonised data template and compound database, always respecting FAIR data storage criteria. For our NAMs we keep track of readiness levels for relevant applications, required for their success and commercial prospects. We also refined the ASPA workflow via interaction with stakeholders (regulators, industry, academia, NGOs) at multiple international events. A major innovation that has started to bring ASPA to the end-users is the NAMASTOX online dashboard to transparently operate on the workflow whilst integrating and documenting the safety assessment procedure.
RH3R established a strong internet presence. Key project publications and events can be found at the project website, in newsletters, and social media.
Lastly, RH3R has continuously provided highly rated training on NGRA to the new generation of scientists on risk assessment, including the practical context of linking lab research and regulatory reality.
The combination of human stem cell-derived tissue models with modern toxicogenomics platforms developed in the project will transform hazard assessment. The combination of this approach, within quantitative systems toxicology models (QSTMs), with our methods to convert exposure information to expected concentration levels in all parts of the body, will be the basis for NGRA of chemicals. Uncertainty measures provided by QSTMs are key to making our output useful for regulatory decision-making.
RH3R is one of three projects constituting the research cluster ASPIS for sustainable, animal-free and reliable chemical risk assessment. Our joint focus is to fulfil regulatory needs whilst allowing for uptake of innovations that promote non-animal safety testing for more reliable human risk assessment. We anticipate that ASPIS, with RH3R as a major contributor, will help solve problems in multiple regulatory contexts, across industry sectors. To achieve the goal of protecting different population groups, ASPIS is now closely engaging with PARC. Important technologies will be shared with this initiative for further implementation.
The major socio-economic impact will be that predictions from our NGRA approach will enable the regulatory definition of acceptable exposure levels, which is required to safeguard public health. As a result, RH3R will have a tangible impact on the development, production and marketing of safer chemical compounds in all sectors by accelerating the toxicity testing process.
RH3R is one of three projects constituting the research cluster ASPIS for sustainable, animal-free and reliable chemical risk assessment. Our joint focus is to fulfil regulatory needs whilst allowing for uptake of innovations that promote non-animal safety testing for more reliable human risk assessment. We anticipate that ASPIS, with RH3R as a major contributor, will help solve problems in multiple regulatory contexts, across industry sectors. To achieve the goal of protecting different population groups, ASPIS is now closely engaging with PARC. Important technologies will be shared with this initiative for further implementation.
The major socio-economic impact will be that predictions from our NGRA approach will enable the regulatory definition of acceptable exposure levels, which is required to safeguard public health. As a result, RH3R will have a tangible impact on the development, production and marketing of safer chemical compounds in all sectors by accelerating the toxicity testing process.