Periodic Reporting for period 1 - ALTERNATIVE (environmentAL Toxicity chEmical mixtuRes through aN innovative platform based on aged cardiac tissue model)
Reporting period: 2021-10-01 to 2023-03-31
There is evidence that cardiovascular diseases relate to environmental exposure to exogenous toxic chemicals, leading to a growing demand for standardised devices and protocols to assess the toxicity of compounds, and standardised procedures to improve chemical safety, and develop reliable risk assessment. In Europe, there are currently no regulatory applications of complex in vitro systems, including bioreactors and Organ-on-Chip. Project ALTERNATIVE aims to develop an in vitro platform able to detect the cardiotoxicity of chemicals and their mixtures. A 3D bio-engineered human functional cardiac tissue will be designed and cultured in a bioreactor system, integrated with multiple sensors. The project focuses on aged cardiac tissue, to evaluate this toxicity in the vulnerable populations, exploiting tissue engineering approaches. It will provide new toxicity markers as tools for early cardiotoxicity detection and monitoring, and a new paradigm for the risk assessment of chemical mixtures, implementing multi-omics analysis and in silico machine learning models. The platform will provide an easy-to-handle, reliable solution reproducing the 3D cardiac tissue and its micro-physiological environment, evaluating the molecular pathways associated with chemical mixtures by omics, implementing in silico machine learning models for cardiotoxicity prediction of chemical mixtures, and identifying and developing new regulatory needs and approaches. By combining in vitro and in silico methods for establishing the toxic profile of chemicals, the platform will by able to reduce and replace animal testing, provide more close-to-real scenario information on toxicity, lower testing costs and result in a significant reduction of the associated direct and indirect costs. The aim of ALTERNATIVE is to design and test an in vitro system mimicking human-aged cardiac tissue to test the toxicity of chemical mixtures and their bio-transformation products through a reliable and high-throughput monitoring system, based on multi-omics analyses and integrated into a machine learning risk assessment tool. In detail the project will develop and provide:
- an integrated system, the ALTERNATIVE platform, for the evaluation of cardiac toxicity induced by chemicals and their mixtures
- an aged cardiac tissue model able to provide a biological response similar to that of living organisms when exposed to toxic chemical mixtures
- omics markers of toxicity through omics analysis to support the development of in silico risk assessment models
- an in-silico cloud computing system for mixture risk assessment
- a proof-of-concept validation of the platform
- stimulate regulatory application and acceptance by providing scientifically-based guidance on how results obtained with the platform can be used to support and evolve regulatory-decision making
- a long-lasting impact on citizens' awareness of the risks related to co-exposure to harmful chemicals and pharmaceuticals
- an integrated system, the ALTERNATIVE platform, for the evaluation of cardiac toxicity induced by chemicals and their mixtures
- an aged cardiac tissue model able to provide a biological response similar to that of living organisms when exposed to toxic chemical mixtures
- omics markers of toxicity through omics analysis to support the development of in silico risk assessment models
- an in-silico cloud computing system for mixture risk assessment
- a proof-of-concept validation of the platform
- stimulate regulatory application and acceptance by providing scientifically-based guidance on how results obtained with the platform can be used to support and evolve regulatory-decision making
- a long-lasting impact on citizens' awareness of the risks related to co-exposure to harmful chemicals and pharmaceuticals
Defined a case study for cardiac toxicity in the aged population
Performed the biological characterization of the 3D cardiac tissue
Performed metabolome, proteome and transcriptome characterization of control and aged cardiac tissues
Obtained list of detected changes in metabolites, proteins or mRNA in chemically treated cardiac tissues and defined possible biomarkers of cardiotoxicity
Obtained list of validated ML models for predicting cardiotoxicity
Deployed an in cloud Big Data Lake for the ML model
ML model running in the data platform
Obtained list of selected substances and mixtures for the case study
Documented regulatory needs and opportunities
Obtained set of novel AOPs for cardiotoxicity
Established dissemination channels
Identified initial list of foreground IPRs, and initial exploitation plans
Performed the biological characterization of the 3D cardiac tissue
Performed metabolome, proteome and transcriptome characterization of control and aged cardiac tissues
Obtained list of detected changes in metabolites, proteins or mRNA in chemically treated cardiac tissues and defined possible biomarkers of cardiotoxicity
Obtained list of validated ML models for predicting cardiotoxicity
Deployed an in cloud Big Data Lake for the ML model
ML model running in the data platform
Obtained list of selected substances and mixtures for the case study
Documented regulatory needs and opportunities
Obtained set of novel AOPs for cardiotoxicity
Established dissemination channels
Identified initial list of foreground IPRs, and initial exploitation plans
In vitro system: deliver a comprehensive bio-engineered 3D cardiac tissue model, which is superior to existing technology in terms of biomimicry of the native microenvironment. The 3D structure can trigger biochemical signals capable of altering protein synthesis and gene transcription that induce changes in proliferation, metabolic rates and gene expression.
Omics analysis: study the effects, at a molecular level, of different chemicals, to predict their impact in the organism, and discover chemical behaviour through toxicokinetic studies. It will design innovative machine learning (ML) in silico tools to explore Adverse Outcome Pathways (AOP).
Computational models: address cardiac toxicity with a quantitative structure–activity relationship of molecular initiating events, to allow the evaluation of the toxicological impact of chemicals on the cardiac tissue when combined in mixtures.
Regulatory assessment: characterise the regulatory needs and structure the generated data according to the AOP framework, thereby facilitating an Integrated Approach to Testing and Assessment for cardiotoxicity.
Gender: consider sex-related variability in cell behaviour, enabling the differentiated evaluation of toxicity pathways and markers.
Develop highly innovative concepts and prototypes to enable the next steps in the assessment of chemical mixtures toxicity, and propose new rules for regulatory assessment.
Expected results and impacts
- Provide an in vitro model for the evaluation of cardiac toxicity in the aged population, evaluating the interactions and the effects of chemicals. Provide a toxicological profile of tested compounds, focusing on mechanisms of action and molecular pathways. Provide a reference system for chemical toxicity analysis of chemicals, replicating physiological cell life conditions. Obtain a list of the changes in metabolites, proteins or mRNA in chemically treated cardiac tissue useful for the analysis of AOPs. This will provide scientific evidence enabling prevention and/or mitigation of co-exposure to pharmaceuticals and industrial chemicals in the environment and the technosphere.
- Introduce a new perspective, supported by in vitro, omics and in silico tools, to cope with co-exposure. The harmful effect arising from the measured co-exposure will be joint to internal exposure models. Describe the approach in a procedure, which can be adopted for regulatory purposes. Focus on fundamental areas of risk, associated with the mortality and morbidity related to adverse heart effects, enabling an efficient risk assessment of substances in a mixture. This will support the implementation of risk assessment and management approaches to reduce critical exposures.
- Address the endpoint of cardiotoxicity, by integrating new approach methodologies, i.e. in silico models, organ-on-a-chip, and omics-based approaches as part of an integrated approach to testing and assessment of cardiotoxicity. As a result, cardiotoxicity can be included as an additional endpoint in regulatory frameworks. This will support the assessment of new regulatory approaches such as mixture assessment factors.
- Develop models to evaluate the cardiotoxicity of pharmaceuticals and contaminants in co-exposure, thereby providing indications useful to better understand and predict adverse activity towards wildlife. Liaise with projects studying ecotoxicity of pharmaceuticals and share relevant information and in-silico models. This will support activities on combined exposures as relevant for the Strategic Approach to Pharmaceuticals in the Environment and as to be defined in the forthcoming Chemical Strategy for Sustainability.
Omics analysis: study the effects, at a molecular level, of different chemicals, to predict their impact in the organism, and discover chemical behaviour through toxicokinetic studies. It will design innovative machine learning (ML) in silico tools to explore Adverse Outcome Pathways (AOP).
Computational models: address cardiac toxicity with a quantitative structure–activity relationship of molecular initiating events, to allow the evaluation of the toxicological impact of chemicals on the cardiac tissue when combined in mixtures.
Regulatory assessment: characterise the regulatory needs and structure the generated data according to the AOP framework, thereby facilitating an Integrated Approach to Testing and Assessment for cardiotoxicity.
Gender: consider sex-related variability in cell behaviour, enabling the differentiated evaluation of toxicity pathways and markers.
Develop highly innovative concepts and prototypes to enable the next steps in the assessment of chemical mixtures toxicity, and propose new rules for regulatory assessment.
Expected results and impacts
- Provide an in vitro model for the evaluation of cardiac toxicity in the aged population, evaluating the interactions and the effects of chemicals. Provide a toxicological profile of tested compounds, focusing on mechanisms of action and molecular pathways. Provide a reference system for chemical toxicity analysis of chemicals, replicating physiological cell life conditions. Obtain a list of the changes in metabolites, proteins or mRNA in chemically treated cardiac tissue useful for the analysis of AOPs. This will provide scientific evidence enabling prevention and/or mitigation of co-exposure to pharmaceuticals and industrial chemicals in the environment and the technosphere.
- Introduce a new perspective, supported by in vitro, omics and in silico tools, to cope with co-exposure. The harmful effect arising from the measured co-exposure will be joint to internal exposure models. Describe the approach in a procedure, which can be adopted for regulatory purposes. Focus on fundamental areas of risk, associated with the mortality and morbidity related to adverse heart effects, enabling an efficient risk assessment of substances in a mixture. This will support the implementation of risk assessment and management approaches to reduce critical exposures.
- Address the endpoint of cardiotoxicity, by integrating new approach methodologies, i.e. in silico models, organ-on-a-chip, and omics-based approaches as part of an integrated approach to testing and assessment of cardiotoxicity. As a result, cardiotoxicity can be included as an additional endpoint in regulatory frameworks. This will support the assessment of new regulatory approaches such as mixture assessment factors.
- Develop models to evaluate the cardiotoxicity of pharmaceuticals and contaminants in co-exposure, thereby providing indications useful to better understand and predict adverse activity towards wildlife. Liaise with projects studying ecotoxicity of pharmaceuticals and share relevant information and in-silico models. This will support activities on combined exposures as relevant for the Strategic Approach to Pharmaceuticals in the Environment and as to be defined in the forthcoming Chemical Strategy for Sustainability.