Presentation of in vivo imaging applied to ecotoxicology

1. In vivo imaging:
a powerful
alternative in
ecotoxicology
Rovaltain
April 8th, 2010
a.briat@animascope.eu

2. ECOTOXICOLOGY
◦ Risk assessment:
‐biological impact of exogenous compounds
‐exposure probability.
◦ REACH: registration, evaluation, authorization and
restrictions of chemicals.
◦ In vivo studies are required to evaluate the risks.
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Ethical impact (large number of animals)
Economical impact (time consuming, high cost)
Rovaltain, April 8th 2010.

3. IMPACT ON THE 3R
Reduce, Refine, Replace
Quantitative, Precise, Predictive
Reduces experimental Artifacts
Longitudinal Studies ‐ Reduce Animal Usage
Reduction and Refinement of animal works are
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Cost and Time‐effective
Need to develop of alternative solutions
= IN VIVO IMAGING 3
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4. Non‐Invasive In Vivo Imaging
Functional Morphological
Imaging Imaging
SPECT US (Echography)
PET CT (X‐rays)
(f)MRI MRI
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Optical
SPECT/CT
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5. Imaging applied to ecotoxicology (1)
Air
hypersensibilisation
pollution
Tissular
Imaging
Aqua Oncogene
toxicity Carcinogene Molecular
imaging
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Geo
toxicity Teratogene
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6. Imaging applied to ecotoxicology (2)
Tissular effect Molecular effect
Inflammation
Imaging of
Embryo development
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gene expression
Cell metabolism
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7. Imagerie de l’inflammation Par IRM
Imaging of inflammation using MRI
acute
inflammation
Chronic 1) Inflammatory nephropathy (Bessaad et al. 2010)
inflammation
T1 T2 T2*
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2) Atherosclerosis
(Lutz et al. Radiology, 2004) 3) Polyarthritis (Hauger et al. JFR; 2006)
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8. Imaging of inflammation using optical imaging
In vivo Near‐InfraRed Fluorescent (NIRF) imaging of
inflammatory joints in the lipopolysaccharide (LPS) induction
model (NIR2‐folate).
(a) White‐light images
(b) NIRF images after NIR2‐folate
injection.
(c) Merged
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(d) H&E‐staining
(e) NIRF images of a longitudinal section
of the LPS‐treated ankles.
Chen, Arthritis Res Ther (2005) 8
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9. Imaging of cell metabolism using
Positron Emission Tomography (PET)
Visualisation of Lymph nodes formation with 18F-FDG PET
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following 140 days TCDD treatment
Vogel et al., Am J Pathology, 2007
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10. Imaging of bone development
using X‐ray Computed Tomography (CT)
30 Influence of diet on
Kinetics of medullary
bone formation and
desorption
BV/TV ( %)
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**
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** :p≤
0 **:p≤0.01, **:0.001 vs d1,
T A B #: p ≤0.05 vs matched BSP+/+
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11. 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
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-Cardiac malformation
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12. Transgenic Animal Models in
Ecotoxicology
Quantitative and real‐time molecular imaging based
on in vivo reporter gene expression:
RE promoter Reporter gene Poly
A
Principle: biosensor genes implicated in tissue damage,
oxydative stress, apoptosis and other cellular pathways.
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‐Optical imaging (Luciferase, GFP)
‐Nuclear imaging (hNIS, HSV‐tk)
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13. Imaging of ionizing radiations
impact on gene expression
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14. Imaging of chemicals exposure
impact on gene expression
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15. 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
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Dusp6 activation
GFP expression
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16. 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.
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REACH the unreachable! 
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17. ANIMASCOPE
One 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 DESIGN
Definition of biological parameters to be measured
Choice of imaging modalities, tracers, biomarkers, contrast agents, animal models
Design of image processing workflow
STUDY MANAGEMENT
Model Pre‐conditioning
Image acquisition
Recording and traceability
IMAGE PROCESSING
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Image handling and pre‐processing
Data analysis
REPORTING
Technical reporting
Biomedical reporting
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18. ANIMASCOPE
Multi‐Modality Imaging
FLEXIBLE and VERSATILE
Optimise 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‐
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invasive technologies, all the main imaging
Imaging facility, Archamps techniques
X‐Ray CT, MRI, PET/SPECT, US, OPTIC
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19. CONTACTS
ANIMASCOPE
ZA Eurekalp, 38660 St.‐Vincent‐de‐Mercuze
(Grenoble) FRANCE
Tel: +33 (0)4 76979487
Fax: +33 (0)9 55753911
www.animascope.eu
info@animascope.eu
New imaging facility Current Animascope office located
near Geneva (74) in Eurekalp (38)
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Rovaltain, April 8th 2010.

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21. Zebra fish embryos as models for embryotoxic and
teratological effects of chemicals
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
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effects of toxicants.
Toxicogenomic expression profiles induced by different toxins
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22. Imaging of toxicity in the
Zebra fish using
in vivo fluorescence imaging
.
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J. Vet. Sci. (2008)
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