Stroke & Alzheimer s Disease: An Inflammatory Duo
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1 Stroke & Alzheimer s Disease: An Inflammatory Duo
2 Stroke and Dementia One in 3 will experience a stroke, dementia or both 64% of persons with a stroke have some degree of cognitive impairment and up to a third have dementia Postmortem: 34% of dementia cases show cerebrovascular pathology Risk factors for cerebrovascular disease are the same for cognitive impairment Prevalence of vascular cognitive impairment (with or without dementia, 2 million in the US)
3 Why Study Stroke & Alzheimer s Together? The combination of silent strokes and low level Alzheimer s results in dementia Nun Study Both share the same risk factors Strokes damaged areas exhibit pathological precursors of Alzheimer s Inflammation is a a dominant force in neurodegenerative properties of stroke and Alzheimer s Clinical stroke and dementia are not treatable Vascular cognitive impairment preceding stroke and Alzheimer s is treatable, i.e. during the brain at risk stage The interactions between small strokes and Alzheimer s Disease during the brain at risk stage (preceding clinical stroke and dementia) may suggest targets for therapy
4 Animal Models β amyloid toxicity (AD): transgenic mice or Amyloid β25-35 icv in the rat Small silent strokes: endothelin injections in the striatum Outcomes AD-like pathology Neuroinflammation cognitive deficits infarct size
5 Why the combination of AD and CI? Alzheimer s APP Pathways Stroke Aβ NFκB Cytokines TNFα, IL-1β Cell Death
6 Rat Model of Cerebral Ischemia in Striatum Endothelin LV 3V Endothelin injections: vasoconstriction
7 Rat Model of Small Striatal Infarcts Ipsilateral p Contralateral AD-like pathology APP Tau-2 OX-6 Neuroinflammation GFAP NFκB
8 Rat Model of β Amyloid Toxicity (AD-like pathology) Aβ (25 35) LV 3V Amyloid β injections: intracerebroventricular
9 β Amyloid Toxicity and APP Immunostaining Anterior Cortex Aβ (25-35) Abeta Control Sham Immunostaining 21 days after bilateral Aβ (25-35) injections Cortex Corpus Callosum Hippocampus
10 β Amyloid Toxicity and new β Amyloid (Αβ immunostaining) in the hippocampus Control Aβ (25-35) 21 days after Aβ (25-35) injections
11 Interactions: Inflammation in the hippocampus Ox6 OX-6 GFAP GFAP Control Levels in the hipocampus 21 days after Aβ (25 35) injections and/or endothelin injections Endothelin (stroke) Aβ (25-35) (Alzheimer s) Aβ and Endo Stroke & Alzheimer s
12 Brain Area Protein Sham Endothelin Aβ Aβ-Endothelin Cortex GFAP OX APP + ++ Tau TNFα + + IL-1β + + NFκB (p65) + + Corpus Callosum GFAP OX APP + ++ Tau TNFα + + IL-1β + + NFκB (p65) + + Thalamus (Right) GFAP OX Hypothalamus GFAP OX TNFα + ++ IL-1β + + NFκB (p65) + ++ Striatum (Right) GFAP OX APP + + Tau TNFα + + IL-1β + + NFκB (p65) + + Hippocampus GFAP OX APP + ++ Tau NFκB (p65) + +
13 Cognitive Testing Barnes Circular Platform Test
14 Circular Platform Test Training Phase 14 trials Surgery test Recovery Period (7, 14 or 28 days)
15 Circular Platform Test Training Phase 14 trials Surgery Recovery Period (7, 14 or 28 days) Test (1 single trial) Re-acquisition (14 trials)
16 Circular Platform Test Sham - Control Training Phase 14 trials Surgery Test Recovery Period (7, 14 or 28 days) Training Re-acquisition 1 single trial Test Re-acquisition 14 trials
17 Endothelin Circular Platform Test Interaction: Aβ and Endothelin Aβ (25-35)
18 Infarct Size Progressive increase in infarct size in presence β amyloid toxicity.
19 Pathways (Alzheimer s Pathway) APP Stroke Aβ NFκB Cytokines TNFα, IL-1β Cell Death
20 NF-κB Activation Pathway PDTC P ΙκB- α Degradation ΙκB- α p50 ΙκB- α p65 X Proteasome IκBkinase p50 p65 p50 p65 Target genes Nucleus mrna Activation signals Cytokines (IL-1β, TNF-α), oxidative stress, Aβ, LPS, phorbol esters, etc. Cytoplasm Inflammatory proteins Inflammatory cytokines (IL-1β, IL- 6, TNF-α), chemokines, adhesion molecules, COX-2, etc
21 Immunostaining of Alzheimer s-related Pathological Markers Sham/Vehicle Aβ (25-35)/Vehicle Aβ (25-35)/PDTC Aβ DG DG DG 100 µm APP DG DG DG DG 100 µm Tau DG DG DG 200 µm
22 Sham/Vehicle Aβ (25-35)/Vehicle Aβ (25-35)/PDTC DG DG DG 100 µm 8 COX-2 Dentate Gyrus Inflammation: COX Reactivity Grade 0 a,b ( ) c ( ) A beta / PDTC A beta / Vehicle Sham / PDTC Sham / Vehicle COX-2
23 Behavioral Testing Results
24 Fluorescent Stain Cerebral Blood Vessels (1) Aβ (FITC) VCAM-1 (TR) Merge Image CBV CBV CBV Aβ 25-35
25 Summary: Animal Models Efficient, reproducible model of Alzheimer-like pathology with progressive neurdegeneration & cognitive impairment Combination beta amyloid toxicity and cerebral ischemia: enhanced pathology, neuroinflammation, cognitive impairment and infarct size NFkappaB inhibition prevents pathological and cognitive deficits Combinations of models required (Alzheimer s with diabetes, hypertension, and atherosclerosis)
26 Vladimir Hachinski Shawn Whitehead John Kiernan Guangliang Cheng
27
28 Preclinical Assessment of New Therapies Anti-inflammatory? Anti-oxidant? Upregulation of gap junctions? Enhanced neurogenesis? Cholinergic (cholinesterase inhibitors)? Amyloid lowering agents?
29 Future Multiple animal models Diverse group of collaborators to assess most of the key mechanisms of neurodegeneration: David Hill, Rob Bartha, Ting Lee, Peter Cain, Edith Arany, David Munoz, Stephen Pasternak, Vladimir Hachinski Clinical affiliation for translational research (Vladimir Hachinski)
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