mixt xtures: focus on pesticides Italian National Institute of - - PowerPoint PPT Presentation
New Approach Methodologies to study effects and modes of f action of f single chemical substances and real mixt xtures: focus on pesticides Italian National Institute of Health General Secretariat Gender-Specific Prevention and Health Unit
Italian National Institute of Health Gender-Specific Prevention and Health Unit Center for Gender-Specific Medicine (MEGE) Toxicology group
76th EFSA AF Meeting 01-02 July 2020
General Secretariat Directorate General for Advisory Bodies for Health Care
Human relevant exposure concentrations
In vitro cell models
Battery of assays
Membrane Potential
Omics
analysis (functional analysis, inference
interacting entities, network analysis)
biomarkers AOP
available
new AOP
Coordinator: ISS-MEGE. «Integrated approach to evaluate children agricultural pesticide exposure and health
chlorpyrifos imidacloprid glyphosate
Literature search for children exposure data MCF7
Range of concentrations: 100 pM – 1 uM
48h 72h
Cytotoxicity assays Gene Expression Hormone secretion
proliferation vitality ERα, ERβ, AR, AhR, PR
MCF12A 2D 3D single or in mixture
Epigenomic profiling
17β-estradiol
In collaboration with Joell Ruegg from Uppsala University
Human mammary gland cells
Coordinator: ISS-MEGE. «In vitro study on potential toxicological effects of active principles and their mixtures used against Peronospora and Oidio.» Funded by Consorzio Vini del Trentino (15/01/2018 – 14/01/2021).
Compound/ Mixtures 1 2 3 4 6 7=18 8=19 9 10=21 12 13 14 15 16 20 Wet sulfur Potassium phosphonate Metrafenone Zoxamide Cyflufenamid Quinoxyfen Mancozeb Folpet Penconazol Dimethomorf
15 Mixtures
Really used concentrations on grapevines (1X)
HepG2
Ten fold dilutions (1 to 106)
24h
Cytotoxicity assays
ROS Mitochondrial Membrane Potential Apoptosis/Necrosis
Gene expression
proliferation vitality
Coordinator: ISS-MEGE. «In vitro study on potential toxicological effects of active principles and their mixtures used against Peronospora and Oidio.» Funded by Consorzio Vini del Trentino (15/01/2018 – 14/01/2021).
A549 BAX, BCL-2 NRF2, CYP1A1
Different modes of action in the two cell lines Collection of the
(IC10, IC20, IC50, Fold change, etc…) Relative toxicological ranking by the ToxPi
Human liver and lung cell lines
Tait et al (2017) Food Chem Toxicol
Transcriptomics Proteomics Functional analysis
In collaboration with the Core Facility at ISS Casella et al (manuscript in preparation)
Polybrominated diphenyl ethers
Bisphenol A effects on placenta angiogenesis
Tait et al (2015) J Appl Toxicol
Systematic review Ad hoc in vitro test development HUVEC primary cells + Human trophoblasts cell line (BeWo) to increase the weight of evidence
In collaboration with the Department of Environment and Health at ISS Members of the EAGMST working group at OECD
compounds allows the identification of subtle and precocious effects which may be harmful for human health
also at environmentally/dietary relevant exposure concentrations
scenario and cross-talks between different cell type for a better definition of adverse effects and modes of action. Particular attention should be given to the use of vitro models considering the two sexes (e.g. primary male and female cells from the same organ) since different effects and modes of action may occur
with different methods, by different groups, gathering all the information in one place. Once an AOP is defined, new methods may be developed for the quantitative weight of evidence of each Key Event/Key Events Relationships.