Alzheimer's disease : Modelization in mouse lemurs
Marc Dhenain
Multimodal Imaging of Neurodegenerative Diseases and Therapies
MIRCen, CEA-CNRS UMR 9199 Fontenay-aux-Roses
CEA | 31 OCTOBER 2014
ALZHEIMER'S DISEASE
Symptoms Dementia
- spatio-temporal disorientation - Alteration of short term memory (episodic) - language, visual recognition
ALZHEIMER'S DISEASE Main cause of dementia
Aging is the first risk factor
ALZHEIMER'S DISEASE CURRENT STATE OF AD DRUG DEVELOPMENT Only five approved drugs (four cholinesterase inhibitors, one NMDA antagonist) 413 trials 124 in Phase 1 206 in Phase 2 83 in Phase 3 Attrition rate of 99.6%!
Data from clinicaltrials.gov looking at period 2002-2012 18 NOVEMBRE 2017 Analysed by Cummings et al. 2014
ALZHEIMER'S DISEASE POST-MORTEM HALLMARKS
β amyloid protein deposits Misfolded proteins
Abnormally phosphorylated Tau proteins
Amyloid plaques Amyloid angiopathy
Tangles Neuropil threads Neuritic corona of the plaques
18 NOVEMBRE 2017 Duyckaerts C et al. Acta Neuropathol. 2009
DIAGNOSTIC OF ALZHEIMER'S DISEASE CONTRIBUTION OF IN-VIVO IMAGING MRI
Cerebral atrophy
Normal aging
Alzheimer Moderate form 40% Temporal atrophy
DIAGNOSTIC OF ALZHEIMER'S DISEASE CONTRIBUTION OF IN-VIVO IMAGING PET
Reduced glucose metabolism
ORIGIN OF AMYLOID PLAQUES
β amyloid protein deposits Misfolded proteins Amyloid plaques Amyloid angiopathy
Beta-secretase
Gamma-secretase
18 NOVEMBRE 2017 APP : Amyloid Protein Precursor
EX. OF AMYLOID PLAQUES FROM APP TO AGGREGATED FORMS OF AMYLOID
Monomeric
Betasecretase
Gammasecretase
soluble Invisible
APP : Amyloid Protein Precursor
soluble ~ Invisible
~Invisible
Visible
STAGES OF AMYLOID DEPOSITION
Thal, D. R., W. S. Griffin and H. Braak (2008). J Cell Mol Med 12(5B): 1848-1862. 18 NOVEMBRE 2017
ALZHEIMER'S DISEASE : FEW GENETIC CAUSES TOWARDS AMYLOID HYPOTHESIS OF ALZHEIMER'S DISEASE
50% of late onset ApoE related
1% of patients
NATURAL HISTORY OF AD BASED ON IMAGING BIOMARKERS ?
Amyloid (PET)
Jack et al, Lancet Neurol. 2013
Dementia
Intensity
Cerebral atrophy Functional alterations (Hypometabolism)
Amyloid plaques (cortex)
Klunk, 2004
Neurofibrillary tangles (cortex)
Mild cognitive impairments
Normal cognition ACADÉMIE
~17 years
NATIONALE DE MÉDECINE, M. DHENAIN – 21 OCTOBRE 2013
Probably wrong
Dementia
~3 yrs ~10 years
ANIMAL MODELS BASED ON AMYLOID HYPOTHESIS OF AD Amyloid precursor Protein (APP)
APP Mutations
ACADÉMIE NATIONALE DE MÉDECINE, M. DHENAIN – 21 OCTOBRE 2013
Amyloide oligomeric (soluble)
Amyloid plaques
ORIGIN OF TAU LESIONS
18 NOVEMBRE 2017
TAU LESIONS PROGRESSIVE COLONISATION OF THE BRAIN
18 NOVEMBRE 2017
Delacourte, A., (1999). Neurology 52(6): 1158-1165.
TAU LESIONS PROGRESSIVE COLONISATION OF THE BRAIN
Braak, H. and E. Braak (1991). Acta Neuropathologica 82: 239-259.
18 NOVEMBRE 2017
Delacourte stade 0 ~ BRAAK STAGE 0
Occipital
External view
Frontal Internal view
Adapté de A. Delacourte
Delacourte stade 1 ~ Braak stage I - Transentorhinal
Trans-entorhinal Cortex Adapté de A. Delacourte
Delacourte stade 2 ~ Braak stage II - Transentorhinal
Entorhinal Cortex (aera 28) Adapté de A. Delacourte
Delacourte stade 3 ~ Braak stage II - Transentorhinal
Mild Cognitive Impairment start
Hippocampus Adapté de A. Delacourte
Delacourte stade 4 ~ Braak stage III - Limbic
MCI probable Slight amyloid deposition
Anterior temporal cortex Adapté de A. Delacourte
Delacourte stade 5 ~ Braak stage III - Limbic
MCI probable ++ Abeta 42 aggregation
Inferior temporal cortex Adapté de A. Delacourte
Delacourte stade 6 ~ Braak stage IV - Limbic
Cognitive alteration start (memory, language, praxies)
Median temporal cortex Adapté de A. Delacourte
Delacourte stade 7 ~ Braak stage V - Isocortical Associative areas language, apraxie
Dementia start
Polymodal associative cortex Adapté de A. Delacourte
Delacourte stade 8 ~ Braak stage V - Isocortical Sensitive or motor associative cortexes
Dementia (slight to moderate)
Broca area
Adapté de A. Delacourte
Delacourte stade 9 ~ Braak stage V - Isocortical Sensitive or motor secondary cortexes Sensitive, visual ou motor primary cortexes
Dementia (moderate to) severe
Adapté de A. Delacourte
Delacourte stade 10 ~ Braak stage VI - Isocortical All the neocortex Several subcortical nuclei
Severe dementia
Adapté de A. Delacourte
TAU LESIONS START IN LOCUS COERULEUS IN CHILDHOOD PROGRESSIVE COLONISATION OF THE BRAIN
11 years
11 years
14 years Braak, Acta Neuropathol, 2011
14 years
14 years
Estimated prevalence of tau pathology
TAU AND AMYLOID OVER AGING 1
0.8
Braak tau stage I-II Braak tau stage III-IV Braak tau stage V-VI Abeta
0.6
0.4
0.2
0 25
50
75
100
Age
Duyckaerts, C. and J. J. Hauw (1997). "Prevalence, incidence and duration of Braak's stages in the general population: can we know?" Neurobiol Aging 18(4): 362-369
CURRENT VIEW OF AD LESIONS
Duyckaerts, C. (2015). Acta Neuropathol 129(5): 749-756.
POSSIBLE INTEGRATED VIEW OF AD PATHOLOGY
Villemagne, V. L. (2015). Lancet Neurology 14(1): 114-124. Concept initially proposed by A. Delacourte
ANIMAL MODELS FOR ALZHEIMER'S DISEASE Alteration of cognitive abilities
Causes ? Mechanisms ? Early diagnosis ? Therapy ? Aging Or
?
Preservation of cognitive abilities
WHAT IS A GOOD ANIMAL ?
Construct validity Biological construction for example (aging…) Genetic construction …
Face validity Phenotypic Endophenotypic Lesions: Amyloide, Tau, Neurodegeneration Endophenotypes accessibles with biomarkers
Prediction validity Cross talk with clinical trials in humans to validate animal models
18 NOVEMBRE 2017
HOW TO FOLLOW-UP ANIMAL MODELS ? Animals
Biomarkers
Humans
Cognitive alterations
?
Cognitive alterations
Atrophy
?
Atrophy
Functional alterations
?
Functional alterations
Amyloid
?
Amyloid
Tau
?
Tau
Translational Research in Neurological Diseases: Biomarkers, M. Dhenain - Feb 2015
MOUSE MODELS BASED ON AMYLOID HYPOTHESIS OF AD Amyloid precursor Protein (APP)
APP Mutations
ACADÉMIE NATIONALE DE MÉDECINE, M. DHENAIN – 21 OCTOBRE 2013
Amyloide oligomeric (soluble)
Amyloid plaques
BEHAVIORAL STUDIES
Rationale: Alzheimer is a dementia Let's look a behavioral alterations in animals to predict drug efficacy…
BEHAVIORAL ALTERATIONS IN RODENTS Ex. Morris water maze
Less time spent in good quadrant in old mice
- Spatial memory (reference memory) - Hippocampal integrity - Widely used
Controls = Tg2576-/-
"Alzheimer" Tg2576+/-
Cognitive alterations But no dementia
Click to see the water maze… Lesne, Nature, 2006
DIFFERENT ORIGIN OF BEHAVIORAL ALTERATIONS IN HUMAN AND ANIMAL Mice
Human
Cognitive alterations Cognitive alterations (not homologous
Cognitive alterations
oligomers
to human alterations)
Homol Prédictive
No Tau
Tau
Amyloid
Amyloid
Translational Research in Neurological Diseases: Biomarkers, M. Dhenain - Feb 2015
Animals
Biomarkers
Humans
Cognitive alterations
Homol Prédictive
Cognitive alterations
Atrophy
?
Atrophy
Functional alterations
?
Functional alterations
Tau
Lack of Tau pathology
Tau
Amyloid
Amyloid
Background
Background
Translational Research in Neurological Diseases: Biomarkers, M. Dhenain - Feb 2015
CEREBRAL ATROPHY IN TRANSGENIC MOUSE MODEL OF AMYLOIDOSIS
Brain and hippocampal growth even in the presence of amyloid deposits… Delatour B. et al.. Neurobiol Aging, 27(6), 835-847; 2006.
Animals
Biomarkers
Humans
Cognitive alterations
Homol Prédictive
Cognitive alterations
Atrophy
Homol Prédictive
Atrophy
Functional alterations
?
Functional alterations
Tau
Lack of Tau pathology
Tau
Amyloid
Amyloid
Background
Background
Translational Research in Neurological Diseases: Biomarkers, M. Dhenain - Feb 2015
Animals
Biomarkers
Humans
Cognitive alterations
Homol Prédictive
Cognitive alterations
Atrophy
Homol Prédictive
Atrophy
Functional alterations
PET/Autorad
Functional alterations
Tau
Lack of Tau pathology
Tau
Amyloid
Amyloid
Background
Background
Translational Research in Neurological Diseases: Biomarkers, M. Dhenain - Feb 2015
CEREBRAL HYPERMETABOLISM IN MOUSE MODELS OF AMYLOIDOSIS
Relative FDG uptake (cortex / cerebellum)
Index of cerebral metabolism (FDG-PET)
Control Amyloid
Poisnel et al, Neurobiol Aging. 2012
Index of micro-metabolism (2DG Autoradiography)
Animals
Biomarkers
Humans
Cognitive alterations
Homol Prédictive
Cognitive alterations
Atrophy
Homol Prédictive
Atrophy
Functional alterations
Homol Prédictive
Functional alterations
Tau
Lack of Tau pathology
Tau
Amyloid
Amyloid
Background
Background
Translational Research in Neurological Diseases: Biomarkers, M. Dhenain - Feb 2015
IMMUNOTHERAPIES IN AMYLOID MICE
Control
Vaccinated (Schenk et al, 1999)
TRANSLATIONAL RESEARCH IN NEUROLOGICAL DISEASES: BIOMARKERS, M.
DISCOVERY OF NEW THERAPY STRATEGIES IN AMYLOID MICE
Control
Vaccinated (Schenk et al, 1999)
TRANSLATIONAL RESEARCH IN NEUROLOGICAL DISEASES: BIOMARKERS, M.
(Holmes et al, 2008)
DISCOVERY OF NEW THERAPY STRATEGIES IN AMYLOID MICE
Control
Vaccinated (Schenk et al, 1999)
No clinical improvement MMSE=0
TRANSLATIONAL RESEARCH IN NEUROLOGICAL DISEASES: BIOMARKERS, M.
(Holmes et al, 2008)
Tau
Neurodegen.
Aβ
Metabolism
TARGET MODELS VERSUS CLINICAL MODELS
Dementia
X Target model Therapy
Predictivity of clinical outcome
Predicting clinical efficacy is impossible with rodent models
Académie Nationale de Médecine, M. Dhenain – 21 Octobre 2013
Bapi
α-sec+
Neurodegen.
Tau
β-sec-
γ-sec-
Immunot.
Aβ
Metabolism
TARGET MODELS ARE USEFUL TO EVALUATE MULTIPLE ANTI-AMYLOID THERAPIES
Target Model
EHT 0202
LY2886721
Avagacestat
Semagacestat
Solanezumab
IVIG
Académie Nationale de Médecine, M. Dhenain – 21 Octobre 2013
Dementia
FIRST TAKE HOME MESSAGE
Do not speak of animal model of Alzheimer's disease
Use a more specific language Model of amyloidosis Target model for amyloidosis
EVALUATION OF OTHER (MORE RELEVANT?) MODELS Small non-humain primate
Microcebus murinus
A FRENCH MODEL FOR CEREBRAL AGING / ALZHEIMER'S DISEASE
1 month
4 years
10 years
Short Longevity : Possible longitudinal follow-up
Small size: Breeding easy, manipulation, experimentation, easy
High reproducibility
1 year
WHY THE MOUSE LEMUR ?
Social life adapted to laboratory
Diffuse amyloid deposits
Young animal
Old animal
Bons et al., Neurobiology of aging, 1991
Limitation: not well known We had to explore everything in this model CARACTERISATION AND VALIDATION OF CEREBRAL AGING IN THE MOUSE LEMUR MODEL
NEUROPATHOLOGY Amyloid precursor protein (APP) similar to that of humans Amyloidosis (10% of old animals)
Tauopathy (rare)
NOVEMBRE 2017 Aging. 2009 Kraska18et al, Neurobiol Picq et al, Neurobiol Aging. 2012
Silhol, Calenda et al., Neurobiol Dis. 1996
Animals
Biomarkers
Humans
Cognitive alterations
?
Cognitive alterations
Atrophy
?
Atrophy
Functional alterations
?
Functional alterations
Tau
Tau
Amyloid
Amyloid
Background
Background
COGNITION NECESSITY TO CREATE COGNITIVE TASKS FOR LEMURS
Jumping stand apparatus Picq JL et al, PLosOne 2015
COGNITION EXAMPLE OF THE JUMPING STAND APPARATUS 0
1 month
Discrimination acquisition
D2 D1
Long-term memory
D1r
18 NOVEMBRE 2017
Picq JL et al, PLosOne 2015
CEA | 31 OCTOBER 2014
OVERVIEW OF COGNITIVE EVALUATIONS IN OLD MOUSE LEMURS
Cognitive deficits in old mouse lemurs
Interindividual variability Vulnerability of some cognitive functions
May remind age-related alteration in humans Procedural memory preserved Early alteration of executive function Declarative memory alteration in a subgroup…
As for mice, aged mouse lemurs do not develop dementia
Animals
Biomarkers
Humans
Cognitive alterations
Homol Prédictive
Cognitive alterations
Atrophy
?
Atrophy
Functional alterations
?
Functional alterations
Tau
Tau
Amyloid
Amyloid
Background
Background
IN VIVO MRI IN MOUSE LEMURS Anesthesia
Monitoring
Isoflurane
Surface Coil
AGE-RELATED IRON ACCUMULATION
Young
Old
CEREBRAL ATROPHY
Young Cd H
Old Cd H
18 NOVEMBRE 2017
EVOLUTION OF CEREBRAL ATROPHY
18 NOVEMBRE 2017
CEREBRAL ATROPHY IN VARIOUS BRAIN REGIONS
Cingulate cortex Occipital Frontal cortex partly spared Subcortical regions such as the caudate or putamen Nucleus basalis of Meynert Nucleus septalis, subiculum, Prepiriform cortex, Hypothalamus
Sawiak et al., Frontiers Aging Neurosc, 2014
QUANTIFICATION OF CEREBRAL ATROPHY
Sawiak et al., Frontiers Aging Neurosc, 2014
HETEROGENEITY 1.9 y
3.0 y
5.3 y
6.9 y
7.6y
1.9 y
3.6y
5.5 y
6.9 y
8.8y
2.1 y
3.6y
2.1 y
2.4y
2.5y
3.9y
5.0 y
5.1 y
5.8 y
5.8 y
5.9 y
6.4 y
6.9 y
6.9 y
7.0 y
7.5 y
10.2 y
10.9 y
11.3y
9.4 y
NORMAL/UNIFORM AGING PROCESS CAUDATE NUCLEUS: ATROPHY OCCURING IN ALL THE AGED ANIMALS
M. Lemur
M. Lemur
3 years
7 years
Picq et al. Neurobiol Aging. 2012
PATHOLOGICAL AGING PROCESS ? HIPPOCAMPUS: ATROPHY OCCURING IN A SUBCATEGORY OF AGED ANIMALS
M. Lemur
M. Lemur
3 years
7 years
A
B
volume hippocampe
(e10 µm 3)
5,0
4,5
4,0
3,5
3,0
2,5 animaux âgés
Picq et al. Neurobiol Aging. PlosOne.
animaux jeunes 2,0 0
1
2
3
4
5
6
âge
7
8
9
10
11
12
13
Functional consequences of atrophy ?
FUNCTIONAL CONSEQUENCES OF ATROPHY ? BEHAVIORAL ALTERATIONS AND ATROPHIED AGED ANIMALS
Correlation between atrophy of septal, hippocampal and entorhinal cortex and age-related cognitive alterations
Only non-human primates showing this pattern Picq et al. Neurobiol Aging. 2012
PATHOLOGICAL AGING PROCESS ? ATROPHY EVOLVING RAPIDLY ?
CSF volume evaluation (arbitrary units)
30 -
20 -
10 -
00
2
4
6
Age (years)
Dhenain et al. Neurobiol Aging. 2000;21(1):81-8.
8
10
PATHOLOGICAL AGING PROCESS ? ATROPHY LEADING TO AMYLOID DEPOSITION ?
CSF volume evaluation (arbitrary units)
30 -
Animals with amyloid deposits Animals without amyloid deposits
20 -
10 -
00
2
4
6
Age (years)
Dhenain et al. Neurobiol Aging. 2000;21(1):81-8.
8
10
APPLICATION SELECTION OF ANIMALS FOR THERAPY TRIALS
Selection of 12 animals Graphe de régression 2,75 2,5
Quantité LCR totale
2,25 2
1,75
1,5
1,25 1 ,75 ,5 ,25 3
3,5
4
4,5
5
5,5 6 Age IRM Y = ,291 + ,191 * X; R^2 = ,09
6,5
7
7,5
8
Animals Cognitive alterations
Biomarkers Homol Prédictive
Atrophy
Functional alterations
Humans Cognitive alterations
Atrophy
?
Functional alterations
Tau
Tau
Amyloid
Amyloid
Background
Background Translational Research in Neurological Diseases: Biomarkers, M. Dhenain - Feb 2015
EVALUATIONS OF GLUCOSE METABOLISM IN LEMURS
Micro-PET Focus220, Siemens Medical solutions Spatial resolution of 1.35 mm Field of view (FOV) of 7.6 cm 3D mode during 60 min Saphene-vein bolus injection 900µCi/100g of [18F]-FDG (CisBio, Orsay, France) Olène Dorieux
AGING LEADS TO CEREBRAL HYPOMETABOLISM
e OB
C
Hypometabolic regions Frontal cortex Hippocampus Cerebellum Caudate O. Dorieux
EVALUATION OF THE EFFECTS OF THERAPIES ON CEREBRAL METABOLISM
Exemple of the use of an IGF1-agonist
Old
Dorieux et al.
SUV
Young
Dorieux O., non publié
EVALUATION OF THE EFFECTS OF THERAPIES ON CEREBRAL METABOLISM Long-chain dietary n-3 polyunsaturated fatty acids from fish oil
Pifferi et al. Journal of Lipid Research, 2015 Dorieux O., non publié
Age : 23±4 months n=6 per group 12 month supplementation
Animals Cognitive alterations
Biomarkers Homol Prédictive
Humans Cognitive alterations
Atrophy
Atrophy
Functional alterations
Functional alterations
Tau
Tau
Amyloid
Amyloid
Background
Background Translational Research in Neurological Diseases: Biomarkers, M. Dhenain - Feb 2015
EXAMPLE OF THERAPY EVALUATIONS
Omega 3 enrichment (Fabien Pifferi, Journal of Lipid Research, 2015, PLoSOne 2011)
Benefits on cognition
Benefits on metabolism
Resveratrol (Dal-Pan A. et al., PLoSOne 2010, Age 2011) IGF1 agonist (Dorieux O., unpublished)
Collaboration avec un grand industriel suisse.
Anti-amyloid immunotherapy (Joseph-Mathurin N., Neurobiology of aging, 2013)
EVALUATION OF THE SIDE EFFECTS OF ANTI-ALZHEIMER IMMUNOTHERAPIES
The first study in human with Aβ1-42 vaccine highlighted severe side effects Cerebral Inflammation
Vaccinated AD patient (Orgogozo et al, 2003)
Microhemorrhages
Patient with microhemorrhages (Viswanathan et al, 2006)
EVALUATION OF THE SIDE EFFECTS OF ANTI-ALZHEIMER IMMUNOTHERAPIES Possibility to follow-up side effects by MRI Injections
MRI
Before
1st
Before
2nd
2 weeks 3 weeks
3rd
6 weeks 7 weeks
+ 2 months
28 weeks
+ 7 months
+ 9 months
4th
42 weeks
Euthanasia
44 weeks
Histology
EVALUATION OF THE SIDE EFFECTS OF ANTI-ALZHEIMER IMMUNOTHERAPIES No sign of inflammation
Before
+2 months
+7 months
+9 months
50 µm Perls iron sections
Joseph-Mathurin et al., Neurobiol Aging, 2013
FOLLOW-UP OF IRON DEPOSITS IN THE CHOROID PLEXUS DURING IMMUNOTHERAPY Before
+9 months
Evolution of total hypointense regions
Volume (mm3) ( Volume
2,00 2.0
Segmentation of the hypointense regions
ANOVA * p
