Ecotox

In vivo In vivo imagingimaging: : a a powerfulpowerful

alternative in alternative in ecotoxicologyecotoxicology

RovaltainRovaltainApril 8April 8thth, 2010, 2010

[email protected]

◦◦

RiskRisk

assessmentassessment::

‐‐biologicalbiological

impact of impact of exogenousexogenous

compoundscompounds

‐‐exposureexposure

probabilityprobability..

◦◦

REACHREACH: registration, : registration, evaluationevaluation, , authorizationauthorization

and and restrictions of restrictions of chemicalschemicals..

◦◦

In vivoIn vivo

studiesstudies

are are requiredrequired

to to evaluateevaluate

the the risksrisks..

EthicalEthical

impact (large impact (large numbernumber

of of animalsanimals) )

EconomicalEconomical

impact (time impact (time consumingconsuming, , highhigh

costcost))

ECOTOXICOLOGYECOTOXICOLOGY

Rovaltain, April 8th 2010.

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IMPACT ON THE 3R Reduce, Refine, Replace

Quantitative, Precise, PredictiveReduces experimental Artifacts

Longitudinal Studies ‐

Reduce Animal Usage

Reduction and Refinement of animal works areCost and Time‐effective

NeedNeed

to to developdevelop

of alternative solutionsof alternative solutions= = IN VIVO IN VIVO IMAGINGIMAGING


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Non‐Invasive In Vivo Imaging

Functional Functional ImagingImaging

SPECTPET

(f)MRIOptical

Morphological Morphological ImagingImaging

US (Echography)CT (X‐rays)

MRI

SPECT/CTSPECT/CT


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Airpollution

Aqua toxicity

Geotoxicity

hypersensibilisation

OncogeneCarcinogene

Teratogene

TissularTissularImagingImaging

MolecularMolecularimagingimaging

Imaging applied to ecotoxicology (1)Imaging applied to ecotoxicology (1)


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Imaging of gene expression

InflammationEmbryo development

Cell metabolism

Imaging applied to ecotoxicology (2)Imaging applied to ecotoxicology (2)

Tissular effectTissular effect Molecular effectMolecular effect


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Imagerie de l’inflammation Par IRM

3) Polyarthritis (Hauger et al. JFR; 2006)2) Atherosclerosis

(Lutz et al. Radiology, 2004)

1) Inflammatory nephropathy (Bessaad et al. 2010)

Imaging of inflammation

using MRI

T1 T2 T2*

acuteinflammation

Chronicinflammation


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(a) White‐light images

(b)

NIRF

images

after

NIR2‐folate injection.

(c) Merged

(d) H&E‐staining

(e)

NIRF

images

of

a

longitudinal

section of the LPS‐treated ankles.

In vivo Near‐InfraRed Fluorescent

(NIRF) imaging of

inflammatory

joints in the lipopolysaccharide (LPS) induction

model (NIR2‐folate).

Imaging of inflammation

using optical imaging

Chen, Arthritis Res Ther (2005)Rovaltain, April 8th 2010.

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Vogel et al., Am J Pathology, 2007

Imaging of Imaging of cell metabolism cell metabolism using using Positron Emission Tomography Positron Emission Tomography (PET)(PET)

Visualisation of Lymph nodes formation with 18F-FDG PET following 140 days TCDD treatment


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TT

3030

2020

1010

00

BV/

TV (

%)

BV/

TV (

%)

AA

****

BB

****

Imaging of Imaging of bone development bone development using using XX‐‐ray Computed Tomographyray Computed Tomography

(CT)(CT)

****:p:p≤≤0.01, 0.01, ****:0.001 vs d1, :0.001 vs d1, #: p #: p ≤≤0.05 vs matched BSP+/+0.05 vs matched BSP+/+

Influence of diet on Influence of diet on Kinetics of medullary Kinetics of medullary bone formation and bone formation and

desorptiondesorption


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Imaging of embryo development

using UltraSonography

(US)

Echography

Combined with Doppler analysis: provides the evaluation of functional parameters, particularly in the cardiovascular and muscular fields:

-Embryo size

-Cardiac malformation


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Transgenic Animal Models in Ecotoxicology

Quantitative and real‐time molecular imaging

based on in vivo reporter gene expression:

Principle:Principle:

biosensor genes implicated in tissue damage, oxydative stress, apoptosis and other cellular pathways..

‐‐Optical imagingOptical imaging

(Luciferase, GFP)(Luciferase, GFP)

‐‐Nuclear imagingNuclear imaging

(hNIS, HSV(hNIS, HSV‐‐tk)tk)

Reporter geneReporter genepromoterpromoter Poly Poly AA

RERE


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Imaging of ionizing radiations Imaging of ionizing radiations impact on gene expressionimpact on gene expression


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Imaging of chemicals exposureImaging of chemicals exposureimpact on gene expressionimpact on gene expression


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Imaging embryotoxic and teratological effects of chemicals in the transgenic zebrafish

Expression of

Dual

Specificity

Phosphatase

6

(dusp6,

also

known

as

Mkp3)

is

controlled

by

FGF

signaling

throughout

development.

The

Dusp6

promoter

drives the expression of GFP in transgenic embryos.

chemical

FGF signaling induction

Dusp6 activation

GFP expression


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Ecotoxicology and Imaging‐

Positive impact on 3R,

‐

Quantitative

and

qualitative

answers

to ecotoxicological questions,

‐

Dedicated animal facility,

‐

Integration of the imaging approach,

‐

Research

programs:

developement

of

new animal models and new methodologies.

REACH the unreachable!


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ANIMASCOPEANIMASCOPE One stop shop for preclinical in vivo imagingOne stop shop for preclinical in vivo imaging

Customizable services and full studies are available depending on customer needs. Every project is

tailored to optimize outcomes and readouts

PROTOCOL DESIGNDefinition of biological parameters to be measuredChoice of imaging modalities, tracers, biomarkers, contrast agents, animal modelsDesign of image processing workflow

STUDY MANAGEMENTModel Pre‐conditioningImage acquisitionRecording and traceability

IMAGE PROCESSINGImage handling and pre‐processingData analysis

REPORTINGTechnical reportingBiomedical reporting


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ANIMASCOPE

Imaging facility, Archamps

Multi‐Modality Imaging

FLEXIBLE and VERSATILEOptimise and Customise Each Protocol

Multi‐modality imaging allows to collect, combine and compare multiple readouts within the same anatomical structure and experimental frame

Animascope utilizes a wide set of non‐ invasive technologies, all the main imaging

techniques

X‐Ray CT, MRI, PET/SPECT, US, OPTIC


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ANIMASCOPEZA Eurekalp, 38660 St.‐Vincent‐de‐Mercuze(Grenoble) FRANCETel: +33 (0)4 76979487Fax: +33 (0)9 [email protected]

CONTACTS

Current Animascope office located in Eurekalp (38)

New imaging facilitynear Geneva (74)


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21

Whole

embryo

microarray studies

yielded

robust

toxicogenomic

profiles

and allowed

to

detect

highly

tissue

restricted

gene responses.

These

response

genes

will be

the

basis

for

the

future

development

of

genetic sensors

of

tissue

specific

effects of toxicants.

Zebra fish embryos as models for embryotoxic and teratological effects of chemicals

Toxicogenomic expression profiles induced by different toxins


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Imaging of toxicity in the Imaging of toxicity in the Zebra fish usingZebra fish using

in vivo in vivo fluorescence imagingfluorescence imaging

J. Vet. Sci. (2008)

.


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