Periodic Reporting for period 3 - OBERON (An integrative strategy of testing systems for identification of EDs related to metabolic disorders)
Okres sprawozdawczy: 2022-01-01 do 2022-12-31
The main objective of the OBERON project is to develop a new battery of tests to detect endocrine disruptors having an impact on metabolic disorders, without the use of animal experimentation. Based on the concept of integrated approach for testing and assessment (IATA), OBERON will combine 1)experimental methods, 2)high throughput omics technologies, 3)epidemiology and human biomonitoring and 4)advanced computational models on functional endpoints related to metabolism.
• WP1 aims at integrating epidemiology and human biomonitoring (HBM) studies with the ED test systems for metabolic disorders (on various time windows: pregnancy-childhood-adolescence and adulthood), in order to increase the relevance of the ED test system.
• WP2’s goal is to develop whole organism test systems to identify EDCs implicated in metabolic disorders (using adiposity, liver steatosis and markers of steatohepatitis as endpoints).
• The global aim of WP3 is to improve/develop innovative in vitro cellular models which best represents the three main organs and tissues (liver, adipose tissue and pancreas) involved in the etiology of metabolic disorders.
• The main objective of WP4 is to develop or adapt existing in silico methods for endocrine disruptor compounds. WP4 will develop QSAR and PK/PBPK models in support of the evaluation and integration of the data streams of the project.
• WP5 aims at integrating information from WP2, WP3 and WP4 in order to interpret the WP1 data (including data infrastructure development, development and application of a bioinformatics workflow of cross-omics and the respective biomarkers towards ED exposure/disease pathways, development of new AOPs relevant EDs outcomes, and development of an integrative testing strategy).
• WP1 aims at integrating epidemiology and human biomonitoring (HBM) studies with the ED test systems for metabolic disorders (on various time windows: pregnancy-childhood-adolescence and adulthood), in order to increase the relevance of the ED test system.
• WP2’s goal is to develop whole organism test systems to identify EDCs implicated in metabolic disorders (using adiposity, liver steatosis and markers of steatohepatitis as endpoints).
• The global aim of WP3 is to improve/develop innovative in vitro cellular models which best represents the three main organs and tissues (liver, adipose tissue and pancreas) involved in the etiology of metabolic disorders.
• The main objective of WP4 is to develop or adapt existing in silico methods for endocrine disruptor compounds. WP4 will develop QSAR and PK/PBPK models in support of the evaluation and integration of the data streams of the project.
• WP5 aims at integrating information from WP2, WP3 and WP4 in order to interpret the WP1 data (including data infrastructure development, development and application of a bioinformatics workflow of cross-omics and the respective biomarkers towards ED exposure/disease pathways, development of new AOPs relevant EDs outcomes, and development of an integrative testing strategy).
WP1- Metabolic health effects of ED exposure in humans: Follow-up and analysis of cohorts were completed. Multi-omics network analysis has been useful to identify several biologically relevant molecular signatures related to non-persistent ED exposure in childhood. Studies in adults have also observed positive associations between PFAS and hepatic enzymes.
WP2 - Disease models using zebrafish: Cell lines stably transfected with zebrafish PPAR evaluated agonist or antagonist activity toward zfPPARγ. A detailed optimized zebrafish obesogenic test (ZOT) protocol was delivered. It has been used to perform the screening of OBERON selected substances in order to determine their obesogenic potential. The optimization of a bioassay to evaluate the potential of selected EDCs to induce liver steatosis (StAZ) has been performed. The screening of selected EDCs for their potential to be induce steatosis has been performed. The chemical analyses of selected compounds during time-course studies of ZOT and StAZ were delivered. In addition, metabolomics data were obtained from samples derived from ZOT as well as transcriptomic analyses of eleuthero-embryos exposed to positive StAZ controls in order to increase toxicological knowledges.
WP3 -2D & 3D models links to organs and tissues: Along the project up to now we have successfully improved and established optimal culture conditions for different liver, adipose and pancreas cellular models. Based on the results of this extensive functional assay, relevant metabolic endpoints were identified and the best cellular model for each organ was selected to be further analysed using high throughput omics technologies. Omics analysis is in progress. In addition, we continue working in the development of validated in vitro test systems to be used for regulatory aspects regarding ED assessment in relation to metabolic disorders.
WP4 - QSAR & PBPK modelling: QSAR models for endocrine disruption (ED) activity and metabolism disruption were reviewed, optimized and will be freely available to the public in the next VEGA release. New models for thyroid peroxidase, the steroidogenesis and glucocorticoid receptor perturbation were developed. A PBPK model for bisphenols in the Zebrafish eleuthero embryo was finalized and showed good predictive capabilities. The human PBPK model has been parameterized for all the compounds and was applied to translate exposure doses into doses in tissues. In addition, biokinetic interactions of co-exposure to 4 phthalates has been elaborated and many metabolic networks have been added to the PBPK model.
WP5 - Data infrastructure, bioinformatics and ITS: Bioinformatics infrastructure was established and analysis of the omics data were delivered. AOP-helpFinder, was updated and added to the AOP-Wiki database. Decision trees based on AI methods were developed for IATA methodology development. ReadEDTest was developed to assess the readiness of in vitro test methods.
WP6 - Project management & coordination: The project Management framework that has been put in place in the first period enables the Project Management Team to closely follow the project activity and timeline. It allows a good reactivity in the day to day management and enables to closely follow the impact of the COVID crisis on the project. OBERON also took actively part in the EURION activities, one webinar on "Omics" has been organized.
WP7 - Dissemination and exploitation, interaction with regulatory bodies: We launched a survey in order to know more about the public’s perception of the EDs and to identify questions on different aspects of this large topic. The survey and the responses collected allowed to build a plan for a filmed interview where an OBERON partner would interview OBERON’s coordinator, like so responding to the general public’s questions on endocrine disruptors.
WP2 - Disease models using zebrafish: Cell lines stably transfected with zebrafish PPAR evaluated agonist or antagonist activity toward zfPPARγ. A detailed optimized zebrafish obesogenic test (ZOT) protocol was delivered. It has been used to perform the screening of OBERON selected substances in order to determine their obesogenic potential. The optimization of a bioassay to evaluate the potential of selected EDCs to induce liver steatosis (StAZ) has been performed. The screening of selected EDCs for their potential to be induce steatosis has been performed. The chemical analyses of selected compounds during time-course studies of ZOT and StAZ were delivered. In addition, metabolomics data were obtained from samples derived from ZOT as well as transcriptomic analyses of eleuthero-embryos exposed to positive StAZ controls in order to increase toxicological knowledges.
WP3 -2D & 3D models links to organs and tissues: Along the project up to now we have successfully improved and established optimal culture conditions for different liver, adipose and pancreas cellular models. Based on the results of this extensive functional assay, relevant metabolic endpoints were identified and the best cellular model for each organ was selected to be further analysed using high throughput omics technologies. Omics analysis is in progress. In addition, we continue working in the development of validated in vitro test systems to be used for regulatory aspects regarding ED assessment in relation to metabolic disorders.
WP4 - QSAR & PBPK modelling: QSAR models for endocrine disruption (ED) activity and metabolism disruption were reviewed, optimized and will be freely available to the public in the next VEGA release. New models for thyroid peroxidase, the steroidogenesis and glucocorticoid receptor perturbation were developed. A PBPK model for bisphenols in the Zebrafish eleuthero embryo was finalized and showed good predictive capabilities. The human PBPK model has been parameterized for all the compounds and was applied to translate exposure doses into doses in tissues. In addition, biokinetic interactions of co-exposure to 4 phthalates has been elaborated and many metabolic networks have been added to the PBPK model.
WP5 - Data infrastructure, bioinformatics and ITS: Bioinformatics infrastructure was established and analysis of the omics data were delivered. AOP-helpFinder, was updated and added to the AOP-Wiki database. Decision trees based on AI methods were developed for IATA methodology development. ReadEDTest was developed to assess the readiness of in vitro test methods.
WP6 - Project management & coordination: The project Management framework that has been put in place in the first period enables the Project Management Team to closely follow the project activity and timeline. It allows a good reactivity in the day to day management and enables to closely follow the impact of the COVID crisis on the project. OBERON also took actively part in the EURION activities, one webinar on "Omics" has been organized.
WP7 - Dissemination and exploitation, interaction with regulatory bodies: We launched a survey in order to know more about the public’s perception of the EDs and to identify questions on different aspects of this large topic. The survey and the responses collected allowed to build a plan for a filmed interview where an OBERON partner would interview OBERON’s coordinator, like so responding to the general public’s questions on endocrine disruptors.
Research conducted in OBERON uses longitudinal epidemiological studies with data on body composition, blood pressure, lipid profiles etc. in addition to applying novel statistical methods. All this to reflect a real-life scenario of effects of EDCs. New and convenient screening test methods for EDCs will be developed for regulatory use, in vivo and in vitro. In silico models feed into the previously mentioned experiments and help guide them contributing to the 3R. A comprehensive and usable IATA will be developed resulting in more efficient public health protection and reducing test chemicals cost as well as socioeconomic impact of ED exposure.