http://www.umcn.nl/Research/Departments/cdl/PRIME/Pages/PRIMELecture.aspx

1. Multi nutrient enriched diets restore cerebral
perfusion and protect against neurodegeneration in a
mouse model for Alzheimer’s disease
Valerio Zerbi1,2, D. Jansen1, X. Fang1, M. Wiesmann1, M. Mutsaers1, P.J. Dederen1,
I.Arnoldussen1, A. Veltien2, S. Van Asten2, A. Heerschap2 and A.J. Kiliaan1
1 Dept. Anatomy, Donders Centre for Neuroscience, Radboud University Nijmegen Medical
Centre, Nijmegen, the Netherlands
2 Dept. Radiology, Radboud University Nijmegen Medical Centre, Nijmegen, the Netherlands

2. Introduction
Alzheimer’s Disease (AD)
AD affects more than 24 million people world wide
Age is the major risk factor for AD: 47% of people older
than 85 years affected
80 million affected in 2040
Clinical phenotype: gradual episodic memory impairment
Neuropathological changes:
• Presence of plaque and tangle pathology
• Massive loss neuronal cells and synapses
• Neurodegeneration / white matter pathology

3. Introduction
Genetics & risk factors
1. Sporadic AD (late onset) Epidemiology (end of 20th century)
Causes unclear Risk factors for Alzheimer’s disease
Risk factors
Ageing
Presence of APOEε4 allele
Hypertension
Congestive heart failure
2. Familial AD (early onset) Atrial fibrillation
Mutations in amyloid precursor Atherosclerosis
Smoking
protein (APP), presenilin (PS)-1
High intake of saturated fat
or -2 genes Diabetes mellitus
contribute to increased Stroke
Sedentary lifestyle
Aβ production Overweight
White matter lesions
De la Torre JC (2002), Stroke

4. Introduction
Regulation of β-amyloid production / clearance
Physiological situation
β-amyloid Microglial cells
γ-secretase
β-secretase
Energy
supply
Blood flow

5. Introduction
Regulation of β-amyloid production / clearance
• Abnormal cleavage of APP by
γ- and β-secretase
β-amyloid Activated
•monomers • Decrease Aβ clearance by
•dimers Microglial cells reduced cerebral blood flow
γ-secretase •trimers
•Aβ oligomers and plaques
β-secretase
• Vascular amyloid deposition
(CAA)
β-amyloid
β-amyloid plaques • Chronic inflammatory
oligomers response
• Energy crisis
Energy
supply
Blood flow • Neurodegeneration

6. Introduction
Regulation of β-amyloid production / clearance
• Abnormal cleavage of APP by
γ- and β-secretase
β-amyloid Activated
•monomers • Decrease Aβ clearance by
•dimers Microglial cells reduced cerebral blood flow
γ-secretase •trimers
•Aβ oligomers and plaques
β-secretase
• Vascular amyloid deposition
(CAA)
β-amyloid
β-amyloid plaques • Chronic inflammatory
oligomers response
• Energy crisis
Energy
supply
Blood flow • Neurodegeneration

7. Introduction
Prevention strategies
• Immunotherapy Targeting Aβ
Still need to prove their efficacy on clinical trials
• “classic” vascular risk factors Prevention vascular disease
(statins, aspirin, NSAIDs) No beneficial effect on Alzheimer pathology
Improving neuronal connectivity
Improvement of neuronal membrane fluidity
• Docosahexaenoic acid (DHA) Improves vascular status and reduces
atherosclerosis
Decrease Aβ levels
Beneficial effect in patients with mild AD #
•*AD2000 collaborative group. Lancet Neurology 2008;7:41-49
•#Freund- levi et al arch neurol 2008

8. Introduction
Prevention strategies
• Immunotherapy Targeting Aβ
Still need to prove their efficacy on clinical trials
• “classic” vascular risk factors Prevention vascular disease
(statins, aspirin, NSAIDs) No beneficial effect on Alzheimer pathology
Improving neuronal connectivity
Improvement of neuronal membrane fluidity
• Docosahexaenoic acid (DHA) Improves vascular status and reduces
atherosclerosis
Decrease Aβ levels
Beneficial effect in patients with mild AD #
•*AD2000 collaborative group. Lancet Neurology 2008;7:41-49
•#Freund- levi et al arch neurol 2008

9. Introduction
Prevention strategies
• Immunotherapy Targeting Aβ
Still need to prove their efficacy on clinical trials
• “classic” vascular risk factors Prevention vascular disease
(statins, aspirin, NSAIDs) No beneficial effect on Alzheimer pathology
Improving neuronal connectivity
Improvement of neuronal membrane fluidity
• Docosahexaenoic acid (DHA) Improves vascular status and reduces
atherosclerosis
Decrease Aβ levels
Beneficial effect in patients with mild AD #
•*AD2000 collaborative group. Lancet Neurology 2008;7:41-49
•#Freund- levi et al arch neurol 2008

10. Aim of the study
Aim of the study
“to investigate the effects of DHA enriched diet on the
pathophysiology of Alzheimer’s Disease”

11. Material and methods
Animal model
• 12 months old male mice
– Transgenic strain resembling familial AD:
APP695swe/PS1∆E9 [APP/PS1]
(Dr. D. Borchelt, Baltimore, MD, USA)
– C57BL/6J [wild type] control mice

12. Material and methods
Diets
Starting from 2 months of age…
(Control) (DHA+) (Fortasyn)
5% soy oil DHA DHA
EPA EPA
Standard UMP UMP
control diet Choline
Phospholipids
B-vitamins
Anti-oxidants
Kamphuis PJ and Scheltens P (2010),
Wurtman RJ (2008)

13. Material and methods
Delatour et al., In Vivo Imaging Biomarkers in Mouse Models of Alzheimer’s
Disease: Are We Lost in Translation or Breaking Through? J Alzh dis 2010

14. 1. 1H MR Spectroscopy

15. Methods – MRS
1H Magnetic Resonance Spectroscopy
Metabolites of interest:
• N-Acetylaspartate (NAA):
neuronal marker,
reflects neuronal dysfunction
• Myo-inositol (mI):
glial marker,
associated with inflammation

16. Results – MRS
Results 1H MRS

17. 2. Cerebral blood flow
(X + CBF) – X = CBF

18. Methods – Cerebral blood flow
FAIR (Flowsensitive alternating inversion recovery)
IR
180 90 180
Slice selective inversion: (X+CBF)
Label slab
Imaging slice
Global inversion: (X)
Global slab
S-G Kim et al, MRM 37:425-435 (1997)
KK Kwong et al, MRM 34:878-887 (1995)

19. Methods – Cerebral blood flow
T1-fitting selective / global
≈ CBF
T1 selective
T1 global
TI (inversion time)

20. Results – Cerebral blood flow
Results – Cerebral Blood Flow

21. 3. Diffusion Tensor Imaging

22. Methods – Diffusion Tensor imaging
Brownian motion
1  (r  r0 )  (r  r0 ) 
Diffusion MR imaging P(r , t | r0 ,0) 
(4Dt )3
exp 

 4 Dt


r 2   6D
• Theoretical result
• Signal attenuation
• Loss of phase coherence
 2  
M xy  M 0 exp     G      D 
  3 

23. Methods – Diffusion Tensor imaging
Anisotropy
• Directionally dependent
• Restriction in diffusion direction
– Because of axonal membrane
• Water molecules diffuse approximately 8 µm
in ~40 ms diffusion time.

24. Methods – Diffusion Tensor imaging
Diffusion tensor imaging
 Dxx Dxy Dxz 
 
• Needs 6 measurements with non co-linear directions D   Dyx Dyy Dyz 
D Dzy Dzz 
 zx 
• Axial diffusivity (λ1)
• Radial diffusivity (RD) [(λ2+ λ3)/2]
• Mean diffusivity (MD)

3 1     
2
2     
2
3   
2
• Fractional anisotropy (FA)

2 1  2  3
2 2 2

http://www.cs.utah.edu/~gk/papers/tvcg00/img144.png
http://www.ajnr.org/content/23/5/803/F6.large.jpg

25. Methods – Diffusion Tensor imaging

26. Results – Diffusion Tensor imaging
Fractional anisotropy (FA, p < 0.05)
Change in APP compared to WT
corpus callosum ventricles optic tract hippocampus
FA ↓↓ ↓ ↓ -

27. Results – Diffusion Tensor imaging
Radial diffusivity (RD, p < 0.05)
Change in APP compared to WT
corpus callosum ventricles optic tract hippocampus
RD ↓ ↑↑ ↑ ↑

28. Summary
Summary
MR Control DHA+ Fortasyn
Hypothesis
Technique diet diet diet
1H metabolic
MRS
alterations
FAIR - ASL Cerebral perfusion
White matter
DT-MRI degeneration and
neuronal loss

29. Acknowledgements
Acknowledgements
Radboud Univ Nijmegen EU consortium 7th framework LipidiDiet
Medical Centre, Nijmegen,
the Netherlands (RUNMC) • Univ of Saarland (USAAR), Germany
Hartmann & Fassbender
Anatomy • Univ of Kuopio (KU), Finland
Amanda Kiliaan Tanila & Soininen
Diane Jansen • Univ of Szeged (USZEG), Hungary
Carola Janssen Penke
Maartje Mutsaers • Tel Aviv University (UTA), Israel
Jos Dederen Michaelson
Ilse Arnoldussen •Göteborgs Universitet (GU), Sweden
Xiaotian Fang Skoog & Gustafson
Maximilian Wiesmann • Danone Research B.V., the Netherlands
Michiel Kleinnijenhuis Broersen
•Karolinska Institutet (KAU), Sweden
Radiology Wahlund
Arend Heerschap •Institute of Physiology (ASCR),
Andor Veltien Czech Republic
Sjaak Van Asten Dolezal
Alan Wright •VU University Medical Centre (VUMC),
the Netherlands
Scheltens
Support: European Community’s Seventh •University of Bonn (UKB), Germany
Framework Programme (FP7/2007-2013) Lütjohann
Under grant agreement no 202167

30. Questions