PEPPER is running and funding the 3 following activities:
Identifying and documenting test methods which are mature, scientifically sound and meeting a need in terms of identification of EDs
Organizing tests on those methods, performed by test laboratories, allowing to assess repeatability and reproducibility of results
Supporting developers in submitting their methods and the required scientific reports to international validation authorities.
Methods overview
The process’s steps :
On-goind methods:
| Year of selection | Phase | Method name | Participating laboratories | Countries | Objective / targets |
| 2020 |  | hPlacentox hPlacenTox-ED testing to evaluate endocrine disruptors and pregnancy disorder (characterisation of placental function using human cell lines) | CNRS, Anses, Eurofins. | FR FR FR | Placental function in pregnant women |
| 2020 |  | LCMS LC-MS/MS based profiling and dynamic modelling of the H295R steroidogenesis assay (measurement of steroid hormones in human adrenal lines) | Rise, VU[1] , Anses, BASF | SE NL FR GE | TG 456 enhancement: 19 hormones (including testosterone, estradiol) involved in: inflammatory response, metabolism, water balance and the development of sexual characteristics, etc. |
| 2020 |  | GRTA Stably transfected human glucocorticoid receptor hGR transcriptional activation assay for detection of agonistic and antagonist activity of chemicals towards hGR (glucocorticoid receptor transactivation in human cell lines) | INSERM, TOXEM, Ineris Tame Water | FR FR FR FR | Functioning of the glucocorticoid response |
| 2021 | | RARTA Stably transfected human retinoic acid receptor RAR transcriptional activation assay for detection of agonistic and antagonist activity of chemicals towards RAR (transactivation of the retinoic acid receptor in human cell lines) | INSERM, TOXEM CRL BfR[2] | FR FR NL GE | Retinoic acid is involved in numerous functions such as reproduction and development, eyesight, etc, |
| 2021 | | AVIAN Avian in ovo assay for sex steroid hormone disrupting properties (sexual development in the Gallus domesticus embryo) | Uppsala U[3] , INRAE, BIONOS | SE FR ES | Sexual differentiation in birds |
| Coupled with method on Quail in Japan. | NIES Japan [4] | JA | |||
| 2021 | | hNPC Human neural progenitor cell proliferation arrest (proliferation of human neural progenitor cells, cultured as floating spheres). | Leibnitz IUF, Oroxcell Uppsala U. | GE FR SE | Neurodevelopmental toxicity |
| 2022 | | In vitro assay for hepatic triglyceride accumulation (measurement of triglyceride accumulation in a HepaRG human liver cell line) | BfR, Oroxcell VITO U Paris Cité | GE FR BE FR | Steatosis (non-alcoholic fatty liver disease), obesity |
| 2022 | | MR TA Stably transfected human mineralocorticoid receptor hMR transcriptional activation assay for detection of agonistic and antagonist activity of chemicals towards hMR (transactivation of the mineralocortoid receptor in a human cell line) | INSERM, TOXEM CRL | FR FR NL | Metabolism of sodium and potassium, disruption of which can lead to oedema and blood pressure problems |
| 2022 | | DIO1 Colorimetric method for assessing deiodinases activity based on Sandell-Kolthoff reaction with human microsomes: DIO1-SK assay (measurement of deiodinase activity on pooled human liver microsomes (cell extracts)) | (BASF and BfR) Pasteur Lille DNTOX[5] | GE GE FR GE | Regulation of thyroid hormone concentration in blood and peripheral tissues |
| 2023 |  | Neurites outgrowth Neurite outgrowth-Neuronal differentiation of C17.2 in response to RAR, RXR and PPARδ (Neurite growth and neuronal differentiation in relation to retinoid and PPARδ pathways in murine cell lines (C17.2)) | Uppsala U Bionos UKHSA5 | SE ES UK | Neurodevelopmental toxicity |
| 2023 | | FITC-TTR Fluorescent FITC-T4 transthyretin competitive binding assay (measurement of the ability of a substance to displace T4 thyroid hormone from its transporter (transthyretin TTR) by labelling T4 with a fluorescein compound (FITC), which reduces the light signal. | (VU) (WFSR[6]) BASF Vito lab Kaly-Cell | NL NL GE BE FR | Transport of thyroid hormones to target organs. Thyroid disruption |
[1] Vrije Universiteit Amsterdam
[2] Bundesinstitut für Risikobewertung, Berlin
[3] University of Uppsala, Sweden.
[4] Japan National Institute for Environmental Studies 16-2 Onogawa Tsukuba, Ibaraki 305-8506 JAPAN
[5] UK Health Security Agency
[6] Wageningen Food Safety Research
PEPPER’s Scientific Council and Relevance Committee held their inaugural meetings respectively on the 5th and 12th June 2020.
The scientific monitoring work which will routinely be done by PEPPER’s team had been entrusted through a call for tender to the ABU consortium, made of 3 teams: Altertox (Belgium), Benaki Phytopathological Institute (Greece) and Paris University.
The consortium conducted a review of existing literature and a series of interviews; they identified and documented more than 12 methods which have been then submitted to the Relevance Committee for examination.
The consortium conducted a review of existing literature and a series of interviews; they identified and documented more than 12 methods which have been then submitted to the Relevance Committee for examination.
The Relevance Committee held its second meeting on 11th September in order to select the first methods to enter the pre-validation process.
Pre-validation work is expected to start early 2021.
PEPPER was presented at :
- the second annual forum on endocrine disruptors of the European Commission, which took place virtually on 17-18 December 2020
- the ARET colloquium-webinar which took place virtually on the afternoons of October 21 and 22, 2020.
Update of test pre-validation
Press release
Launch of two new methods.