Periodic Reporting for period 2 - ALTERNATIVE (environmentAL Toxicity chEmical mixtuRes through aN innovative platform based on aged cardiac tissue model)
Período documentado: 2023-04-01 hasta 2025-03-31
The Problem Being Addressed
Cardiovascular diseases are increasingly linked to environmental exposure to toxic chemicals. Yet Europe lacked standardised devices, protocols, and regulatory applications to reliably assess chemical mixture toxicity. Traditional animal-based testing cannot capture complex mixture effects and poorly reflects human physiology, especially in vulnerable groups such as the elderly. There was therefore a clear need for innovative, human-relevant tools to improve chemical safety and risk assessment.
Importance for Society
- Cardiovascular diseases remain a leading cause of morbidity and mortality worldwide.
- Citizens are exposed to mixtures of chemicals and pharmaceuticals, often without clear knowledge of long-term risks.
- Improved testing tools can reduce reliance on animal models, lower costs, and accelerate regulatory decisions.
- By focusing on aged cardiac tissue, the project directly addresses vulnerable populations, ensuring risk assessments are inclusive and protective.
The societal benefits include stronger public health protection, greater consumer safety, and enhanced trust in regulatory systems, while reinforcing Europe’s leadership in sustainable and ethical innovation.
Overall Objectives
The ALTERNATIVE project aimed to design and validate an in vitro system mimicking human-aged cardiac tissue to test the toxicity of chemical mixtures and their biotransformation products. This was achieved through a high-throughput monitoring system based on multi-omics analyses and integrated into a machine learning risk assessment tool. Specifically, the project delivered:
- An integrated system (ALTERNATIVE platform) for evaluating cardiac toxicity of chemicals and mixtures.
- An aged cardiac tissue model reproducing biological responses of living organisms.
- Omics markers of toxicity to support mechanistic understanding and in silico modelling.
- An in-silico cloud system for mixture risk assessment, hosting validated ML models and Big Data pipelines.
- Proof-of-concept validation against selected chemicals and mixtures.
- Regulatory guidance and stimulation of acceptance, including novel AOPs, IATA concepts, and validation roadmaps.
- Long-lasting impact through dissemination, exploitation, and raising citizen awareness of chemical co-exposure risks.
Conclusions of the Action
The project successfully delivered a validated, human-relevant platform combining in vitro and in silico methods for cardiotoxicity testing. It demonstrated that:
- The platform can reduce and replace animal testing while providing more realistic toxicity data.
- It enables early detection of cardiotoxicity through novel biomarkers and omics analyses.
- It supports regulatory innovation, with AOPs submitted to OECD and guidance documents drafted for uptake.
- Dissemination and exploitation ensured visibility to regulators, industry, and the scientific community, while IPR management and exploitation plans pave the way for future use.
Overall, ALTERNATIVE introduced a new paradigm for chemical mixture risk assessment, with significant scientific, regulatory, socio-economic, and societal impacts. It strengthens Europe’s position in advancing Next Generation Risk Assessment (NGRA) and contributes to safer, more sustainable chemical management.
Cardiovascular diseases are increasingly linked to environmental exposure to toxic chemicals. Yet Europe lacked standardised devices, protocols, and regulatory applications to reliably assess chemical mixture toxicity. Traditional animal-based testing cannot capture complex mixture effects and poorly reflects human physiology, especially in vulnerable groups such as the elderly. There was therefore a clear need for innovative, human-relevant tools to improve chemical safety and risk assessment.
Importance for Society
- Cardiovascular diseases remain a leading cause of morbidity and mortality worldwide.
- Citizens are exposed to mixtures of chemicals and pharmaceuticals, often without clear knowledge of long-term risks.
- Improved testing tools can reduce reliance on animal models, lower costs, and accelerate regulatory decisions.
- By focusing on aged cardiac tissue, the project directly addresses vulnerable populations, ensuring risk assessments are inclusive and protective.
The societal benefits include stronger public health protection, greater consumer safety, and enhanced trust in regulatory systems, while reinforcing Europe’s leadership in sustainable and ethical innovation.
Overall Objectives
The ALTERNATIVE project aimed to design and validate an in vitro system mimicking human-aged cardiac tissue to test the toxicity of chemical mixtures and their biotransformation products. This was achieved through a high-throughput monitoring system based on multi-omics analyses and integrated into a machine learning risk assessment tool. Specifically, the project delivered:
- An integrated system (ALTERNATIVE platform) for evaluating cardiac toxicity of chemicals and mixtures.
- An aged cardiac tissue model reproducing biological responses of living organisms.
- Omics markers of toxicity to support mechanistic understanding and in silico modelling.
- An in-silico cloud system for mixture risk assessment, hosting validated ML models and Big Data pipelines.
- Proof-of-concept validation against selected chemicals and mixtures.
- Regulatory guidance and stimulation of acceptance, including novel AOPs, IATA concepts, and validation roadmaps.
- Long-lasting impact through dissemination, exploitation, and raising citizen awareness of chemical co-exposure risks.
Conclusions of the Action
The project successfully delivered a validated, human-relevant platform combining in vitro and in silico methods for cardiotoxicity testing. It demonstrated that:
- The platform can reduce and replace animal testing while providing more realistic toxicity data.
- It enables early detection of cardiotoxicity through novel biomarkers and omics analyses.
- It supports regulatory innovation, with AOPs submitted to OECD and guidance documents drafted for uptake.
- Dissemination and exploitation ensured visibility to regulators, industry, and the scientific community, while IPR management and exploitation plans pave the way for future use.
Overall, ALTERNATIVE introduced a new paradigm for chemical mixture risk assessment, with significant scientific, regulatory, socio-economic, and societal impacts. It strengthens Europe’s position in advancing Next Generation Risk Assessment (NGRA) and contributes to safer, more sustainable chemical management.
- ALTERNATIVE platform implemented: 3D scaffold, GelMA hydrogel, co-culture system, refined bioreactor, disposable electrode frames, QC methods.
- Case study defined for aged population cardiotoxicity (chemicals list, 7 day exposure).
- Recommendations produced for mixture risk assessment (AOP-based IATA).
- 3D cardiac tissue characterized (PU scaffold, GelMA hydrogels, functionalization, cytocompatibility, multi-omics).
- Toxicological profiling completed for selected chemicals in static young/aged models.
- Reference system established for chemical toxicity analysis in static conditions.
- Omics characterization performed (metabolome, proteome, transcriptome of young/aged tissues).
- Detected changes listed in metabolites, proteins, mRNA; biomarkers of cardiotoxicity defined (finalization ongoing).
- Derivative compounds fully characterized from toxic chemical metabolization.
- TK models deployed; TD adapted to statistical approaches.
- Validated ML models (QSAR) predicting cardiotoxicity and biological target interactions.
- Cloud Big Data Lake implemented for ML input/output.
- Data pipelines integrated for ingestion and processing.
- ML models operational in platform, supporting risk assessment.
- Case study substances/mixtures selected (mitochondrial effects).
- ML model validation parameters achieved against chemicals/mixtures.
- New cardiac markers evaluated from omics data.
- Validated against in silico models and open data; case study outcome achieved.
- Regulatory needs documented.
- Novel/existing AOPs defined (OECD AOP Wiki submission).
- Draft guidance prepared for regulatory use (tiered IATA).
- EURL-ECVAM pre-submission form and roadmap completed.
- Dissemination channels established.
- Foreground IPRs continuously updated.
- Individual and joint exploitation plans developed and refined.
- Case study defined for aged population cardiotoxicity (chemicals list, 7 day exposure).
- Recommendations produced for mixture risk assessment (AOP-based IATA).
- 3D cardiac tissue characterized (PU scaffold, GelMA hydrogels, functionalization, cytocompatibility, multi-omics).
- Toxicological profiling completed for selected chemicals in static young/aged models.
- Reference system established for chemical toxicity analysis in static conditions.
- Omics characterization performed (metabolome, proteome, transcriptome of young/aged tissues).
- Detected changes listed in metabolites, proteins, mRNA; biomarkers of cardiotoxicity defined (finalization ongoing).
- Derivative compounds fully characterized from toxic chemical metabolization.
- TK models deployed; TD adapted to statistical approaches.
- Validated ML models (QSAR) predicting cardiotoxicity and biological target interactions.
- Cloud Big Data Lake implemented for ML input/output.
- Data pipelines integrated for ingestion and processing.
- ML models operational in platform, supporting risk assessment.
- Case study substances/mixtures selected (mitochondrial effects).
- ML model validation parameters achieved against chemicals/mixtures.
- New cardiac markers evaluated from omics data.
- Validated against in silico models and open data; case study outcome achieved.
- Regulatory needs documented.
- Novel/existing AOPs defined (OECD AOP Wiki submission).
- Draft guidance prepared for regulatory use (tiered IATA).
- EURL-ECVAM pre-submission form and roadmap completed.
- Dissemination channels established.
- Foreground IPRs continuously updated.
- Individual and joint exploitation plans developed and refined.
The ALTERNATIVE project advanced toxicological science by creating the first integrated in vitro aged cardiac tissue model combined with multi-omics profiling and machine learning risk assessment. This marks a major step beyond the state of the art, moving from animal-based testing and single-compound analysis toward human-relevant, mechanistic, and mixture-focused approaches. Key innovations include:
- A bi-phasic scaffold system mimicking young and aged cardiac tissue, validated for cytocompatibility.
- Multi-omics integration (metabolomics, proteomics, transcriptomics) to identify biomarkers of cardiotoxicity.
- A cloud-based ML platform ingesting large-scale omics and toxicological data to predict mixture effects and support regulatory decisions.
- Novel Adverse Outcome Pathways (AOPs) for cardiotoxicity, submitted to OECD for endorsement.
Results of the Project
- Platform implementation and validation: System deployed, tested with chemicals and mixtures, validated against omics markers and ML models.
- Scientific outputs: Biomarkers identified, TK models developed, ML models validated, derivative compounds characterized.
- Regulatory outputs: Guidance documents, AOPs, and validation roadmaps produced, supporting acceptance of non-animal methods.
- Exploitation and dissemination: Channels established, IPRs identified and updated, exploitation plans developed among partners.
Potential Impacts
Scientific and Technological
- Establishes a new paradigm for cardiotoxicity testing that is scalable and human-relevant.
- Provides a validated alternative to animal testing, aligned with EU goals on New Approach Methodologies (NAMs).
- Delivers a cloud-based ML risk assessment tool extendable to other organ systems.
Socio-Economic
- Reduces reliance on costly animal testing, lowering development costs for industry.
- Accelerates innovation in safety assessment, enabling faster approvals and safer products.
- Strengthens Europe’s global leadership in NAMs.
Societal
- Improves public health protection by detecting cardiotoxic risks in chemical mixtures, especially for the elderly.
- Raises citizen awareness of chemical exposure risks through dissemination.
- Supports policy innovation, advancing EU ambitions for sustainable chemicals management and international frameworks.
- A bi-phasic scaffold system mimicking young and aged cardiac tissue, validated for cytocompatibility.
- Multi-omics integration (metabolomics, proteomics, transcriptomics) to identify biomarkers of cardiotoxicity.
- A cloud-based ML platform ingesting large-scale omics and toxicological data to predict mixture effects and support regulatory decisions.
- Novel Adverse Outcome Pathways (AOPs) for cardiotoxicity, submitted to OECD for endorsement.
Results of the Project
- Platform implementation and validation: System deployed, tested with chemicals and mixtures, validated against omics markers and ML models.
- Scientific outputs: Biomarkers identified, TK models developed, ML models validated, derivative compounds characterized.
- Regulatory outputs: Guidance documents, AOPs, and validation roadmaps produced, supporting acceptance of non-animal methods.
- Exploitation and dissemination: Channels established, IPRs identified and updated, exploitation plans developed among partners.
Potential Impacts
Scientific and Technological
- Establishes a new paradigm for cardiotoxicity testing that is scalable and human-relevant.
- Provides a validated alternative to animal testing, aligned with EU goals on New Approach Methodologies (NAMs).
- Delivers a cloud-based ML risk assessment tool extendable to other organ systems.
Socio-Economic
- Reduces reliance on costly animal testing, lowering development costs for industry.
- Accelerates innovation in safety assessment, enabling faster approvals and safer products.
- Strengthens Europe’s global leadership in NAMs.
Societal
- Improves public health protection by detecting cardiotoxic risks in chemical mixtures, especially for the elderly.
- Raises citizen awareness of chemical exposure risks through dissemination.
- Supports policy innovation, advancing EU ambitions for sustainable chemicals management and international frameworks.