Assays for the identification of Thyroid Hormone axis-disrupting chemicals: Elaborating Novel Assessment strategies

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Deliverables

Report on effects of thyroid hormone axis-disrupting chemicals on human-specific thyroid physiology including thyroidal stimulation by hCG

Report on the effects of thyroid hormone axis disruptingchemicals on humanspecific thyroid physiology including thyroidal stimulation by hCG We will detail the outcome of activities in Task 92

Policy briefing no 2 to the European Commission and JRC
Provisional list of identified transmembrane transporters in placenta, BBB and BCSFB

Provisional list of identified transmembrane transporters in placenta BBB and BCSFB prioritized as candidate proteins for new screening assays month 30 ERASMUS STICHTING VUVUA This will detail known transporters to be selected as candidate proteins for new screening assays

Qualitative AOP network of the thyroid hormone axis for adverse neurodevelopmental outcomes

Qualitative AOP network of the thyroid hormone axis for adverse neurodevelopmental outcomes This deliverable will describe the outcome of Task 101

Risk assessment procedure
Concept for interaction with other H2020 projects on EDC test method development

Concept for interaction with other H2020 projects on EDC test method development This will detail a series of activities with the other 7 projects funded under this call

Policy briefing no 1 to the European Commission and JRC

Policy briefing no 1 to the European Commission and JRC: These policy briefing will summarise relevant work in ATHENA, together with policy implications

Mediation analyses on the identified thyroid hormone axis-disrupting chemicals to quantify how much of their effects is mediated via thyroid hormone axis

Perform mediation analyses on the identified THADCs to quantify how much of their effect on neurocognitive outcomes is mediated via thyroid function (month 66) (ERASMUS, KAU): We will describe the outcome of mediation analyses

Report on the role of TTR in transport of thyroid hormone across in vitro placenta and BCSFB models

Report on the role of TTR in transport of thyroid hormones and EDCs across in vitro placenta and BCSFB models (month 66; VUA): This deliverable will detail the role of TTR in the transport of thyroid hormones across the placenta and the BCSFB.

Initial setup of DEHAL1 screening assay and pilot screen cellular membrane transporter with an EDC compound library

A stable cell line clone recombinantly overexpressing human DEHAL1 at sufficient activity levels required for implementation of the HTS will be used in the DEHAL1 enzyme activity test based on the nonradioactive SandellKolthoff reaction A specific DEHAL1 inhibitor will be used for quality control in assay development A small EDC compound library will be tested as a pilot screen for the cellular membrane transporter MCT8 in a 96 well format The aim is to identify potent MCT8 inhibitors

Analysis of commonalities and differences in screening, testing and hazard/risk assessment of substances covered by case studies

Analysis of commonalities and differences in screening testing and hazardrisk assessment of substances covered by case studies

Policy briefing no 3 to the European Commission and JRC
Options for creating institutional platforms for international hazard and risk assessment for thyroid hormone axis-disrupting chemicals
Standard Operating Procedures v 1.0 for test methods developed in WP 4-7

Standard Operating Procedures v 1.0 for test methods developed in WP 4-7: We will take forward all test methods from WP 4-7 in terms of technical SOP as described in Task 8.1

Report on newly identified transmembrane transporters in placenta, BBB and BCSFB

Report on newly identified transmembrane transporters in placenta, BBB, and BCSFB containing the ultimate prioritized list of candidate proteins for new screening assays (month 66; ERASMUS, STICHTING VUVUA): This deliverable will list newly identified transporters and the ultimate list of candidate proteins for novel screening assays

Report on the screening of approx. 50 chemicals identified in WP 1 and 4

Report on the screening of approximately 50 chemicals identified in WP1 and 4 (month 69): We will share the experiences from the testing of ca. 50 chemicals with the new platform detailed in D 7.1

Pilot DEHAL1 screen with an EDC compound library and final HTS of cellular membrane transporter with large chemical libraries for small molecules

A small EDC compound library will be tested as a pilot screen DEHAL1 in a 96 well format The aim is to identify potent DEHAL1 inhibitors The final HTS MCT8 of the cellular membrane transporter MCT8 with large libraries for small molecules in the 384 well format will be implemented together with subcontractor FMP We expect completion of this final MCT8 T3 transporter HTS by month 45 and expect identification of new potent and specific MCT8 inhibitors Curated data of the completed final MCT8 T3 transport HTS will be transferred to our WP2 partners for development of QSAR models by month 45

Document inventory of screening and testing activities specifically for thyroid hormone axis-disrupting chemicals, including priority setting principles and risk assessments

Document inventory of screening and testing activities specifically for thyroid hormone axisdisrupting chemicals including priority setting principles and risk assessments and overview summary of the document inventory D 111

Identification of thyroid hormone axis-disrupting chemicals for which consequences on gestational thyroid function are influenced by iodine status

Identification of thyroid hormone axisdisrupting chemicals for which consequences on gestational thyroid function are influenced by iodine status This deliverable will describe the outcome of Task 93

Final HTS DEHAL1 and DIO2 of large chemical libraries for small molecules and report on suggested reference compound libraries for inhibition of deiodinases, dehalogenases, and cell membrane transporters

The final HTS DEHAL1 and DIO2 screen of large libraries of small molecules in the 384 well format will be implemented together with our subcontractor FMP. On the basis of results and achievements of D10, D11 and D12 we expect completion of these final HTS DEHAL1 and DIO2 screens by month 63 and expect identification of new potent and specific DEHAL1 as well as DIO2 inhibitors. Curated data of the completed final DEHAL1 and DIO2 HTS will be transferred to our WP2 partners for development of QSAR models.The report on suggested reference compound libraries of this deliverable will draw together a) the results of the biochemical final HTS for the two enzymes DIO2 and DEHAL1 (D4.4) with b) our previous results from the biochemical final HTS for Deiodinase 3 (DIO3) in comparison to the c) results of the cellular membrane transporter MCT8 final HTS (D4.3) using the same Sandell-Kolthoff assay platform and the same large chemical libraries of small molecules. The identified inhibitors for each target will be compared with respect to potential overlaps or selectivity of inhibition. This will reveal the list of reference compounds for each EDC target screened and for the planned in vivo testing in intact animal models of the other WPs. The extensive inhibitor data sets obtained in WP4 provide the experimental basis for the generation of training and validation sets for QSAR modelling developed by DTU partner 2 in WP2.

Potential of upscaling and refinement for each of the models

"Potential of upscaling and refinement for each of the models (month 60, CNRS): #This will describe and analyse the potential of the above 3D models for upscaling for routine testing"

Preliminary plan for exploitation and dissemination

Release of a preliminary plan for exploitation and dissemination

Protocol for a cell-based screening assay with over-expressed newly identified transmembrane transporters

Protocol for a cell-based screening assay with overexpressed newly identified transmembrane thyroid hormone transporter (depending on D 5.1) (month 66; VUA): We will provide a description of screening assays with newly identified transporters, selected on the basis of D 5.1 and D 5.2

Extended version of XETA test guideline

Extended version of XETA test guideline (month 62). The deliverable will describe the outcome of tasks 4-6

Final conference
Standard Operating Procedures v 2.0, incorporating the outcome of in-house validations

Standard Operating Procedures v 2.0, incorporating the outcome of in-house validations: This will describe the outcomes of in house validations as detailed in Task 8.2

Report on cellular and molecular data on consequences of the interactions between iodine deficiency and thyroid hormone axis-disrupting chemicals on brain development

Report on cellular and molecular data on consequences of the interactions between iodine deficiencyhypothyroidism and THADCs on brain development in two vertebrate models mouse and xenopus This deliverable will describe the activities in Task 91

High through-put cell based screening assay with overexpressed OATP1C1

High throughput cellbased screening assay with overexpressed OATP1C1 month 36 VUA CHARIT A description of HTP screening assay with overexpressed OATP1C1

Validate and replicate novel findings on newly identified thyroid disruptors to human thyroid function tests using epidemiological data

Validate and replicate novel findings on newly identified thyroid disruptors to human thyroid function tests using human epidemiological data (month 66) (ERASMUS): Data on newly discovered thyroid disruptors will be evaluated by using existing or newly emerged epidemiological studies

Policy briefing no 4 to the European Commission and JRC
Data on associations of exposure to major EDC subgroups with maternal thyroid function during pregnancy

Provide data on the association of exposure to major EDCs subgroups with maternal thyroid function during pregnancy (month18) (ERASMUS): We will leverage epidemiological studies with information on EDC exposures and T4 levels in pregnancy to provide criteria for test compound selection in the subsequent WP

Final plan for exploitation and dissemination

A final plan for exploitation ana dissemination will be detailed and released based on the project outcomes.

QSAR models for inhibition of DIO1, DIO2, DIO3 and TSHR

QSAR models for inhibition of DIO1, DIO2, DIO3 and TSHR: This will detail the characteristics of new QSAR models

Reporting templates and project handbook

Reporting templates and project handbook: we will create reporting templates and a project handbook, to be delivered to all partners to ease the reporting on finances and deliverables

Report on the calibration and identification of thyroid specific and sensitive endpoints in 3D cell models

Report on the calibration and identification of thyroid hormone specific and sensitive endpoints in each of the 3D cell based models month 24 CNRS and CHARIT We will provide a description of new thyroid sensitive endpoints in 3D cell culture models

Final testing protocol for inclusion of endpoints of thyroid hormone disruption in the developing brain that can be incorporated into OECD TG 443 and other relevant guidelines

Final testing protocol for inclusion of endpoints of thyroid hormone disruption in the developing brain that can be incorporated into OECD TG 443 and other relevant guidelines (month 66): The preliminary testing protocol in D 6.1 will be finalised, with detailed descriptions of positive control substances, timing of exposure, route of administration, harvesting of tissues and number of animals to be used

Definition of endpoints characteristic of effects of chemicals on human brain organoids and mouse neurospheres

Definition of endpoints characteristic of effects of suspected and established thyroid hormone disruptive chemicals on human brain organoids and mouse neurospheres (month 60, CNRS and CHARITÉ): We will detail and define relevant endpoints in human brain organoids and mouse neurospheres

Integrated testing strategy for thyroid hormone axis-disrupting chemicals

Integrated testing strategy for thyroid hormone axis-disrupting chemicals: This deliverable will describe an integrated testing strategy, the outcome of work conducted in Task 10.3

Preliminary testing protocol for inclusion of measurement endpoints of thyroid hormone action that can be incorporated into relevant OECD test guidelines

Preliminary testing protocol for inclusion of measurement endpoints of thyroid hormone action that can be incorporated into relevant OECD test guidelines month 36 This will describe endpoints for downstream effects of thyroid disruption on the brain together with a preliminary testing protocol

Final Release of new QSARs in free public websites
Project website

Project website The site will link to a portal of all projects funded under this call and will detail the work programme and partners in ATHENA

Release of new QSARs in free public websites

Release of the new QSARs in free public websites month 54 The new models will be released in websites

Workshop 2 with international advisory panel
Workshop 3 with international advisory panel
Workshop 1 with international advisory panel

Workshop 1 with international advisory panel month 30

Data Management plan

Submission of a first version of a Data Management Plan (DMP) at month 6 of the project. The DMP will be updated over the course of the project whenever significant changes arise, such as (but not limited to): - new data - changes in consortium policies (e.g. new innovation potential, decision to file for a patent) - changes in consortium composition and external factors (e.g. new consortium members joining or old members leaving).

A robotised screening platform for the XETA

A robotized screening platform for the XETA (month 60): This will describe all technical aspects of the robotized screening platform

Publications

A Mixture of Chemicals Found in Human Amniotic Fluid Disrupts Brain Gene Expression and Behavior in Xenopus laevis.

Author(s): Leemans M, Spirhanzlova P, Couderq S, Le Mével S, Grimaldi A, Duvernois-Berthet E, Demeneix B, Fini JB
Published in: International Journal of Molecular Sciences, Issue 24, 2023, ISSN 1422-0067
Publisher: Multidisciplinary Digital Publishing Institute (MDPI)
DOI: 10.3390/ijms24032588

Developmental thyroid disruption permanently affects the neuroglial output in the murine subventricular zone.

Author(s): Vancamp, P., Le Blay, K., Butruille, L., Sébillot, A., Boelen, A., Demeneix, B. A., & Remaud, S
Published in: Stem Cell Reports, Issue 17, 2022, ISSN 2213-6711
Publisher: Cell Press
DOI: 10.1016/j.stemcr.2022.01.002

Two thyroperoxidase-inhibiting chemicals induce shared transcriptional changes in hippocampus of developing rats

Author(s): Louise Ramhøj, Terje Svingen, Bertrand Evrard, Frédéric Chalmel, Marta Axelstad
Published in: Toxicology, Issue 505, 2025, Page(s) 153822, ISSN 0300-483X
Publisher: Elsevier BV
DOI: 10.1016/j.tox.2024.153822

Perinatal exposure to environmental chemicals that disrupt thyroid function can perturb testis development

Author(s): Monica K. Draskau, Ida W. Strand, Raul Ayala Davila, Anne-Sofie R. Ballegaard, Mikael Pedersen, Louise Ramhøj, Sofie Rising, Kieu-mi Tran, Marta Axelstad, Josephine Bowles, Anna K. Rosenmai, Cassy M. Spiller, Terje Svingen
Published in: Environmental Pollution, Issue 363, 2024, Page(s) 125117, ISSN 0269-7491
Publisher: Pergamon Press Ltd.
DOI: 10.1016/j.envpol.2024.125117

Association of urinary bisphenols and triclosan with thyroid function during early pregnancy

Author(s): Arash Derakhshan, Huan Shu, Robin P. Peeters, Andreas Kortenkamp, Christian H. Lindh, Barbara Demeneix, Carl-Gustaf Bornehag, Tim I.M. Korevaar
Published in: Environment International, Issue 133, 2019, Page(s) 105123, ISSN 0160-4120
Publisher: Pergamon Press Ltd.
DOI: 10.1016/j.envint.2019.105123

Perinatal exposure to the thyroperoxidase inhibitors methimazole and amitrole perturbs thyroid hormone system signaling and alters motor activity in rat offspring.

Author(s): Ramhøj L, Svingen T, Frädrich C, Rijntjes E, Wirth EK, Pedersen K, Köhrle J, Axelstad M
Published in: Toxicology Letters, 2021, ISSN 0378-4274
Publisher: Elsevier BV
DOI: 10.1016/j.toxlet.2021.10.010

Mechanistic insights into the synergistic activation of the RXR-PXR heterodimer by endocrine disruptor mixtures.

Author(s): Delfosse V, Huet T, Harrus D, Granell M, Bourguet M, Gardia-Parège C, Chiavarina B, Grimaldi M, Le Mével S, Blanc P, Huang D, Gruszczyk J, Demeneix B, Cianférani S, Fini JB, Balaguer P, Bourguet W.
Published in: Proceedings of the National Academy of Sciences, Issue 118, 2020, ISSN 1091-6490
Publisher: National Academy of Sciences
DOI: 10.1073/pnas.2020551118

Association of endocrine disrupting chemicals exposure with human chorionic gonadotropin concentrations in pregnancy.

Author(s): Derakhshan A, Shu H, Broeren MAC, Kortenkamp A, Lindh CH, Demeneix B, Peeters RP, Bornehag CG, Korevaar TIM
Published in: Environment International, Issue 178, 2023, ISSN 0160-4120
Publisher: Pergamon Press Ltd.
DOI: 10.1016/j.envint.2023.108091

Testing for thyroid hormone disruptors, a review of non-mammalian in vivo models

Author(s): Stephan Couderq, Michelle Leemans, Jean-Baptiste Fini
Published in: Molecular and Cellular Endocrinology, Issue 508, 2020, Page(s) 110779, ISSN 0303-7207
Publisher: Elsevier BV
DOI: 10.1016/j.mce.2020.110779

Two novel in vitro assays to screen chemicals for their capacity to inhibit thyroid hormone transmembrane transporter proteins OATP1C1 and OAT4

Author(s): Fabian Wagenaars, Peter Cenijn, Zhongli Chen, Marcel Meima, Martin Scholze, Timo Hamers
Published in: Archives of Toxicology, Issue 98, 2024, Page(s) 3019-3034, ISSN 0340-5761
Publisher: Springer Verlag
DOI: 10.1007/s00204-024-03787-2

Transient developmental exposure to low doses of bisphenol F negatively affects neurogliogenesis and olfactory behaviour in adult mice,

Author(s): Vancamp, P., Butruille, L., Herranen, A., Boelen, A., Fini, J. B., Demeneix, B. A., & Remaud, S.
Published in: Environment International, Issue 172, 2023, ISSN 0160-4120
Publisher: Pergamon Press Ltd.
DOI: 10.1016/j.envint.2023.107770

Removing Critical Gaps in Chemical Test Methods by Developing New Assays for the Identification of Thyroid Hormone System-Disrupting Chemicals—The ATHENA Project

Author(s): Andreas Kortenkamp, Marta Axelstad, Asma H. Baig, Åke Bergman, Carl-Gustaf Bornehag, Peter Cenijn, Sofie Christiansen, Barbara Demeneix, Arash Derakhshan, Jean-Baptiste Fini, Caroline Frädrich, Timo Hamers, Lina Hellwig, Josef Köhrle, Tim I.M. Korevaar, Johan Lindberg, Olwenn Martin, Marcel E. Meima, Philipp Mergenthaler, Nikolai Nikolov, David Du Pasquier, Robin P. Peeters, Bjorn Platzack, Lou
Published in: International Journal of Molecular Sciences, Issue 21/9, 2020, Page(s) 3123, ISSN 1422-0067
Publisher: Multidisciplinary Digital Publishing Institute (MDPI)
DOI: 10.3390/ijms21093123

Association of phthalate exposure with thyroid function during pregnancy

Author(s): Arash Derakhshan, Huan Shu, Maarten A.C. Broeren, Christian H. Lindh, Robin P. Peeters, Andreas Kortenkamp, Barbara Demeneix, Carl-Gustaf Bornehag, Tim I.M. Korevaar.
Published in: Environment International, Issue 157, 2021, Page(s) 106795, ISSN 0160-4120
Publisher: Pergamon Press Ltd.
DOI: 10.1016/j.envint.2021.106795

Association of per- and polyfluoroalkyl substances with thyroid homeostasis during pregnancy in the SELMA study

Author(s): Derakhshan A, Kortenkamp A, Shu H, Broeren MAC, Lindh CH, Peeters RP, Bornehag CG, Demeneix B, Korevaar TIM
Published in: Environment International, Issue 167, 2022, ISSN 0160-4120
Publisher: Pergamon Press Ltd.
DOI: 10.1016/j.envint.2022.107420

Testing for heterotopia formation in rats after developmental exposure to selected in vitro inhibitors of thyroperoxidase.

Author(s): Ramhøj L, Frädrich C, Svingen T, Scholze M, Wirth EK, Rijntjes E, Köhrle J, Kortenkamp A, Axelstad M.
Published in: Environmental Pollution, 2021, ISSN 0269-7491
Publisher: Pergamon Press Ltd.
DOI: 10.1016/j.envpol.2021.117135

Asymmetrical Transport of Thyroxine Across Human Term Placenta

Author(s): Chen Z, Peeters RP, Leeuwenburgh S, Broekhuizen M, Neuman RI, Hitzerd E, Tan L, Jongejan RMS, de Rijke YB, Reiss IKM, Danser AHJ, Visser WE, Meima ME
Published in: Thyroid, Issue 33, 2021, ISSN 1050-7256
Publisher: Mary Ann Liebert Inc.
DOI: 10.1089/thy.2022.0406

Distinct transcriptional profiles in rat thyroid glands after developmental exposure to three in vitro thyroperoxidase inhibiting chemicals

Author(s): Anna Kjerstine Rosenmai, Terje Svingen, Bertrand Evrard, Khanh Hoang Nguyen, Camilla Nielsen, Marta Axelstad, Frédéric Chalmel, Louise Ramhøj
Published in: Genomics, Issue 116, 2024, Page(s) 110938, ISSN 0888-7543
Publisher: Academic Press
DOI: 10.1016/j.ygeno.2024.110938

Pesticides With Potential Thyroid Hormone-Disrupting Effects: A Review of Recent Data

Author(s): Michelle Leemans, Stephan Couderq, Barbara Demeneix, Jean-Baptiste Fini
Published in: Frontiers in Endocrinology, Issue 10, 2019, ISSN 1664-2392
Publisher: Frontiers
DOI: 10.3389/fendo.2019.00743

Comparative analysis of thyroid hormone systems in rodents with subterranean lifestyle.

Author(s): Gerhardt P, Begall S, Frädrich C, Renko K, Hildebrandt TB, Holtze S, Heinrich A, Sahm A, Meci X, Köhrle J, Rijntjes E, Henning Y
Published in: Nature Scientific Reports, Issue 13, 2023, ISSN 2045-2322
Publisher: Nature Publishing Group
DOI: 10.1038/s41598-023-30179-w

Deiodinases control local cellular and systemic thyroid hormone availability.

Author(s): Köhrle J, Frädrich C
Published in: Free Radical Biology and Medicine, Issue 193, 2022, ISSN 0891-5849
Publisher: Elsevier BV
DOI: 10.1016/j.freeradbiomed.2022.09.024

Is periventricular heterotopia a useful endpoint for developmental thyroid hormone system disruption in mouse toxicity studies?

Author(s): Louise Ramhøj, Romain Guyot, Terje Svingen, Andreas Kortenkamp, Frédéric Flamant, Marta Axelstad
Published in: Regulatory Toxicology and Pharmacology, Issue 142, 2024, Page(s) 105445, ISSN 0273-2300
Publisher: Academic Press
DOI: 10.1016/j.yrtph.2023.105445

Characterization of thyroid hormone transport in a human choroid plexus papilloma cell line (HIBCPP) as an in vitro blood-cerebrospinal fluid barrier model

Author(s): Fabian Wagenaars, Peter Cenijn, Jeske van Boxel, Jacco Koekkoek, Horst Schroten, Hiroshi Ishikawa, Majorie van Duursen, Timo Hamers
Published in: Molecular and Cellular Endocrinology, Issue 597, 2025, Page(s) 112449, ISSN 0303-7207
Publisher: Elsevier BV
DOI: 10.1016/j.mce.2024.112449

Novel (sulfated) thyroid hormone transporters in the solute carrier 22 family.

Author(s): Chen Z, Peeters RP, Flach W, de Rooij LJ, Yildiz S, Teumer A, Nauck M, Sterenborg RBTM, Rutten JHW, Medici M, Edward Visser W, Meima ME
Published in: European Thyroid Journal, Issue 12, 2023, ISSN 2235-0640
Publisher: S. Karger AG
DOI: 10.1530/etj-23-0023

Selenium, Iodine and Iron-Essential Trace Elements for Thyroid Hormone Synthesis and Metabolism.

Author(s): Köhrle J
Published in: International Journal of Molecular Sciences, Issue 24, 2023, ISSN 1422-0067
Publisher: Multidisciplinary Digital Publishing Institute (MDPI)
DOI: 10.3390/ijms24043393

Thyroid Hormone Transporters in a Human Placental Cell Model

Author(s): Chen Z, van der Sman ASE, Groeneweg S, de Rooij LJ, Visser WE, Peeters RP, Meima ME
Published in: Thyroid, Issue 32, 2022, ISSN 1050-7256
Publisher: Mary Ann Liebert Inc.
DOI: 10.1089/thy.2021.0503

Developmental exposure to thyroid disruptors: misprogramming of the brain's stem cells in later life?

Author(s): Vancamp P, Remaud S.
Published in: Neural Regen Res, Issue 18, 2023, ISSN 1673-5374
Publisher: Neural Regeneration Research
DOI: 10.4103/1673-5374.346053

Association of urinary bisphenols during pregnancy with maternal, cord blood and childhood thyroid function

Author(s): Arash Derakhshan, Elise M Philips, Akhgar Ghassabian, Susana Santos, Alexandros G Asimakopoulos, Kurunthachalam Kannan, Andreas Kortenkamp, Vincent WV Jaddoe, Leonardo Trasande, Robin P Peeters, Tim IM Korevaar
Published in: Environment International, Issue 146, 2021, Page(s) 106160, ISSN 0160-4120
Publisher: Pergamon Press Ltd.
DOI: 10.1016/j.envint.2020.106160

Screening for endocrine disrupting chemicals inhibiting monocarboxylate 8 (MCT8) transporter facilitated thyroid hormone transport using a modified nonradioactive assay

Author(s): Fabian Wagenaars, Peter Cenijn, Martin Scholze, Caroline Frädrich, Kostja Renko, Josef Köhrle, Timo Hamers
Published in: Toxicology in Vitro, Issue 96, 2024, Page(s) 105770, ISSN 0887-2333
Publisher: Elsevier BV
DOI: 10.1016/j.tiv.2023.105770

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