1CMEP Plenary Meeting - Prague, 30/06/2011

SETAC Europe 21st Annual Meeting, Milan

Joint Research CentreInstitute for Environment and Sustainabilityhttp://ies.jrc.ec.europa.eu/

CMEP Task 3.2 – Effect based monitoring tools

“ Omics” technologies towards the

water quality risk assessment for

environmental monitoring

Raquel Negrão CarvalhoTeresa Lettieri

2CMEP Plenary Meeting - Prague, 30/06/2011

SETAC Europe 21st Annual Meeting, Milan

Mode of Action (MoA): chemical-specific molecular signature

• Omics approaches, including DNA microarray are used to determine MoA of priority substances in model organisms of relevant trophic levels

Unaffected by treatmentUp-regulated by treatment

Down-regulated by treatment

3CMEP Plenary Meeting - Prague, 30/06/2011

SETAC Europe 21st Annual Meeting, Milan

Test species

At beginning the sequencing was concentrated on pathogens or species relevant for biomedical field

Many other organisms relevant in environment field were sequenced

Caulobacter crescentus (Bacteria)

Sea squirt (Eukaryota)

Fugu fishDiatom (Eukaryota)

Daphnia pulex (Eukaryota)

rainbow trout

Zebrafish

fathead minnow

Chlamydomonas (green algae)

Synechoccocus (cyanobacteria)

4CMEP Plenary Meeting - Prague, 30/06/2011

SETAC Europe 21st Annual Meeting, Milan

Whole systems approaches to environmental assessment

Perturbation•Pollutants•Physical stressors•Locations

OMICS•Metabolome•Proteome•Transcriptome•Interactions

Organism

Acute Endpoints(EC/LC50)•Survival•Reproduction•Development

Available bioassays•Vitelogenin•CYP450•Androgen receptor•Estrogen receptor•…

Pathway inference

Co-regulation discovery

MoA

Cellular reporters/Biomarkers

Bioassay

Identify hazardClasses of pollutants

5CMEP Plenary Meeting - Prague, 30/06/2011

SETAC Europe 21st Annual Meeting, Milan

Molecular based bioassay Saccharomyces cerevisiae Unicellular eukaryote

Highly conserved with larger eukaryotes including mammalsNot pathogenic easy to handle Does not require any biosafety level

Cytotoxicity analysis (selected compounds )

Up regulated

Down regulatedNo change

Selection of the mutant strain

• More sensitivity to the specific class of compounds.• Easy approach to generate mutants in the same strain to target different compounds

6CMEP Plenary Meeting - Prague, 30/06/2011

SETAC Europe 21st Annual Meeting, Milan

1μM Flu.Molecular-based Bioassay

Wild-type

S288c

W303.1a

RAD1

YGR131w

Control 1 μM Flu.103105 104 102 10 103105 104 102 10

3μM Flu.103105 104 102 10

Development of more sensitive strainse.g. Knock-out of genes involved in Oxidative stress response

7CMEP Plenary Meeting - Prague, 30/06/2011

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- Molecular level response could be linked to a dose-response and allow sample classification regarding environmental hazard

- Omics approaches allow exposure of environmental sample without pre-treatment/ fractionation steps - reduced costs, assessment of chemical mixtures and new substances

- Different models could be used by different laboratories (if MoA known and biomarkers have been validated for that system)

- Information on MoA allows the development of even more sensitive tools

FINAL REMARKS

8CMEP Plenary Meeting - Prague, 30/06/2011

SETAC Europe 21st Annual Meeting, Milan

Thank you!