Periodic Reporting for period 4 - GOLIATH (Beating Goliath: Generation Of NoveL, Integrated and Internationally Harmonised Approaches for Testing Metabolism Disrupting Compounds)
Période du rapport: 2023-07-01 au 2024-12-31
The incidence of metabolic disorders like obesity, diabetes, and fatty liver disease has reached alarming levels. While lifestyle and genetics play a role, environmental factors—especially metabolism-disrupting chemicals (MDCs), a subset of endocrine-disrupting chemicals (EDCs)—are increasingly recognized as contributors. MDCs, both natural and synthetic, can drive metabolic changes leading to these diseases. To address their role, international regulations must mandate MDC identification and risk assessment.
European EDC regulations require evidence of endocrine mode of action (MoA) and adverse effects, but no regulatory in vivo or in vitro tests exist for MDCs, hindering risk assessment.
The GOLIATH project (www.beatinggoliath.eu) improves EDC risk assessment by developing novel, harmonized approaches for testing metabolic disruption. It spans in silico modeling, high-throughput screening, in vitro assay development, and optimization of in vivo testing guidelines. By integrating omics technologies, GOLIATH links assay outcomes to human health, offering new insights into MDC mechanisms. With a multidisciplinary expert consortium, GOLIATH advances a globally harmonized MDC testing strategy.
The overall objectives of the project are:
• To improve the understanding of the endocrine modes of action of MDCs.
• To develop assay candidates for metabolic disrupting chemicals based on confirmed MoA and key biological effects in target tissues.
• To select and develop assay candidates into (pre-)validated test methods, in collaboration with OECD, ensuring test method definition, transferability, inter-laboratory testing and assessment of predictivity, which are prerequisites for their regulatory use.
• To develop an internationally harmonised, integrated approach to testing and assessment (IATA) of MDCs, using an Adverse Outcome Pathway (AOP) conceptual framework.
European EDC regulations require evidence of endocrine mode of action (MoA) and adverse effects, but no regulatory in vivo or in vitro tests exist for MDCs, hindering risk assessment.
The GOLIATH project (www.beatinggoliath.eu) improves EDC risk assessment by developing novel, harmonized approaches for testing metabolic disruption. It spans in silico modeling, high-throughput screening, in vitro assay development, and optimization of in vivo testing guidelines. By integrating omics technologies, GOLIATH links assay outcomes to human health, offering new insights into MDC mechanisms. With a multidisciplinary expert consortium, GOLIATH advances a globally harmonized MDC testing strategy.
The overall objectives of the project are:
• To improve the understanding of the endocrine modes of action of MDCs.
• To develop assay candidates for metabolic disrupting chemicals based on confirmed MoA and key biological effects in target tissues.
• To select and develop assay candidates into (pre-)validated test methods, in collaboration with OECD, ensuring test method definition, transferability, inter-laboratory testing and assessment of predictivity, which are prerequisites for their regulatory use.
• To develop an internationally harmonised, integrated approach to testing and assessment (IATA) of MDCs, using an Adverse Outcome Pathway (AOP) conceptual framework.
The main results from the GOLIATH project include the following achievements, many of which have been published and are available on our website (https://beatinggoliath.eu/publications/)(s’ouvre dans une nouvelle fenêtre):
• The current state of science and overview of human exposure to MDCs published in a brief review and further elaborated as a detailed review paper within the OECD programme.
• Advanced in silico screening models developed for MDC interaction prediction, interaction of MDCs with AhR, PXR and RXR elucidated, EDMON platform enhanced (http://atome.cbs.cnrs.fr/ATOME_V3/SERVER/EDMon_v3.html)(s’ouvre dans une nouvelle fenêtre);
• Advanced in vitro assay development: PPAR gamma activity characterization, hepatocyte assays (steatosis, insulin resistance, CYP induction assay, pancreatic alpha and beta cell assays, adipocyte differentiation assays
• Pre-validation of in vitro assays including chemical selection for the PPAR gamma and alpha, CYP induction, and adipocyte differentiation assays
• Metabolic profiling approaches for the understanding of the endocrine modes of action of MDCs, including the development of the DEXOM algorithm for cell-specific metabolic network characterization, and FORUM for chemical-health data mining.
• Multi-omics analysis of gene expression, metabolomics and lipidomics in in vitro, zebrafish and human models
• Development of the proteome-based thermal shift assay for MDCs in zebrafish, hepatocytes, and adipocytes.
• In vivo method development of the transgenic zebrafish model for MDCs
• Epidemiological analysis of anthropogenic measures and markers in cohorts relative to prenatal MDC exposure Multi-omics analysis of gene expression, metabolomics and lipidomics in in vitro, zebrafish and human models.
• IATA development: Mechanistic networks developed for obesity, diabetes and fatty liver. Framework for IATA developed (publication in progress, will be further elaborated in PARC).
• Weight of evidence assessment for MDCs elaborated in expert elicitation approach
• Augmentation of test guidelines for how to include metabolic disruption measures in animal study guidelines.
• Collaboration with EURION partners (https://eurion-cluster.eu/(s’ouvre dans une nouvelle fenêtre)) within working groups and international outreach within EURION through stakeholder meetings which led to more awareness of GOLIATH achievements and steps towards regulatory acceptance.
In conclusion, the GOLIATH project has developed and pre-validated multiple test methods and approaches for the identification of MDCs. As a result, GOLIATH has contributed significantly to the scientific state of the art and emerging concepts in the field of MDCs and MDC testing.
• The current state of science and overview of human exposure to MDCs published in a brief review and further elaborated as a detailed review paper within the OECD programme.
• Advanced in silico screening models developed for MDC interaction prediction, interaction of MDCs with AhR, PXR and RXR elucidated, EDMON platform enhanced (http://atome.cbs.cnrs.fr/ATOME_V3/SERVER/EDMon_v3.html)(s’ouvre dans une nouvelle fenêtre);
• Advanced in vitro assay development: PPAR gamma activity characterization, hepatocyte assays (steatosis, insulin resistance, CYP induction assay, pancreatic alpha and beta cell assays, adipocyte differentiation assays
• Pre-validation of in vitro assays including chemical selection for the PPAR gamma and alpha, CYP induction, and adipocyte differentiation assays
• Metabolic profiling approaches for the understanding of the endocrine modes of action of MDCs, including the development of the DEXOM algorithm for cell-specific metabolic network characterization, and FORUM for chemical-health data mining.
• Multi-omics analysis of gene expression, metabolomics and lipidomics in in vitro, zebrafish and human models
• Development of the proteome-based thermal shift assay for MDCs in zebrafish, hepatocytes, and adipocytes.
• In vivo method development of the transgenic zebrafish model for MDCs
• Epidemiological analysis of anthropogenic measures and markers in cohorts relative to prenatal MDC exposure Multi-omics analysis of gene expression, metabolomics and lipidomics in in vitro, zebrafish and human models.
• IATA development: Mechanistic networks developed for obesity, diabetes and fatty liver. Framework for IATA developed (publication in progress, will be further elaborated in PARC).
• Weight of evidence assessment for MDCs elaborated in expert elicitation approach
• Augmentation of test guidelines for how to include metabolic disruption measures in animal study guidelines.
• Collaboration with EURION partners (https://eurion-cluster.eu/(s’ouvre dans une nouvelle fenêtre)) within working groups and international outreach within EURION through stakeholder meetings which led to more awareness of GOLIATH achievements and steps towards regulatory acceptance.
In conclusion, the GOLIATH project has developed and pre-validated multiple test methods and approaches for the identification of MDCs. As a result, GOLIATH has contributed significantly to the scientific state of the art and emerging concepts in the field of MDCs and MDC testing.
GOLIATH has progressed beyond the state of the art and contributed to the following four expected impacts mentioned in the work programme:
1)Improved hazard and risk assessment of endocrine disruptors (EDs), including in the workplace.
Currently, no test guidelines exist to assess MDC hazard and risk. GOLIATH has developed the first integrated approach to testing and assessment (IATA) for MDCs, including standardized, pre-validated methods. We pre-validated three human in vitro assays (PPARγ/PPARα reporter gene and hMSC adipogenesis) and enhanced the CYP induction assay. Combined with metabolomics and lipidomics, these methods provide critical insights into chemical properties and MoA by modeling human metabolic network shifts after exposure.
2)Novel ED assay candidates for regulatory use.
GOLIATH aimed to identify and confirm mechanisms of action for new ED assay candidates. We expanded in silico screening tools for MDC-nuclear receptor interactions (e.g. QSAR, EdMon) and enhanced web interfaces for metabolic MoA prediction (MetExplore) to support regulators. Additionally, we optimized and applied thermal protein profiling to MDCs, identifying numerous direct and indirect protein targets.
3)Enhanced international cooperation.
GOLIATH has supported OECD efforts on MDC testing. Our project plan for a detailed review paper (DRP) on metabolic disruption was accepted by the OECD WNT in April 2020, with the first version undergoing review. Through UKHSA, GOLIATH contributed to OECD discussions on enhancing the CYP induction test and will advance it toward Test Guideline acceptance.
4)Contribution to the development of an international strategy and guidelines for testing EDs.
GOLIATH integrated all data to establish the first conceptual IATA for MDCs, linking human exposure to key mechanisms. This approach provides a mechanistic understanding of MDC contributions to obesity, type 2 diabetes, and fatty liver disease. By combining novel in vitro assays, in vivo models, and human data, it maps disrupted biological processes. This IATA forms the foundation for further development in projects like PARC and EDC-MASLD.
1)Improved hazard and risk assessment of endocrine disruptors (EDs), including in the workplace.
Currently, no test guidelines exist to assess MDC hazard and risk. GOLIATH has developed the first integrated approach to testing and assessment (IATA) for MDCs, including standardized, pre-validated methods. We pre-validated three human in vitro assays (PPARγ/PPARα reporter gene and hMSC adipogenesis) and enhanced the CYP induction assay. Combined with metabolomics and lipidomics, these methods provide critical insights into chemical properties and MoA by modeling human metabolic network shifts after exposure.
2)Novel ED assay candidates for regulatory use.
GOLIATH aimed to identify and confirm mechanisms of action for new ED assay candidates. We expanded in silico screening tools for MDC-nuclear receptor interactions (e.g. QSAR, EdMon) and enhanced web interfaces for metabolic MoA prediction (MetExplore) to support regulators. Additionally, we optimized and applied thermal protein profiling to MDCs, identifying numerous direct and indirect protein targets.
3)Enhanced international cooperation.
GOLIATH has supported OECD efforts on MDC testing. Our project plan for a detailed review paper (DRP) on metabolic disruption was accepted by the OECD WNT in April 2020, with the first version undergoing review. Through UKHSA, GOLIATH contributed to OECD discussions on enhancing the CYP induction test and will advance it toward Test Guideline acceptance.
4)Contribution to the development of an international strategy and guidelines for testing EDs.
GOLIATH integrated all data to establish the first conceptual IATA for MDCs, linking human exposure to key mechanisms. This approach provides a mechanistic understanding of MDC contributions to obesity, type 2 diabetes, and fatty liver disease. By combining novel in vitro assays, in vivo models, and human data, it maps disrupted biological processes. This IATA forms the foundation for further development in projects like PARC and EDC-MASLD.