F.J. Gonzalez-Velasquez, J.W. Reed, J.W. Fuseler, J.A. Kotarek, D.D. Soto-Ortega, and M.A. Moss
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Transcript of F.J. Gonzalez-Velasquez, J.W. Reed, J.W. Fuseler, J.A. Kotarek, D.D. Soto-Ortega, and M.A. Moss
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Activation of endothelium in Alzheimer’s brain involves soluble aggregates of the amyloid-protein
F.J. Gonzalez-Velasquez, J.W. Reed,J.W. Fuseler, J.A. Kotarek,
D.D. Soto-Ortega, and M.A. Moss
University of South Carolina
College of Engineering and ComputingUniversity of South Carolina
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College of Engineering and ComputingUniversity of South Carolina
Alzheimer’s Pathology
Alzheimer’s Brain
Pathology
A Fibrils
Nichols et al. (2002)Biochemistry, 41: 6115-27.
Courtesy of D. Dickson,Mayo Clinic, Jacksonville.
Cerebral AmyloidAngiopathy
http://www.adrc.wustl.edu/adrc2.html
Amyloid Plaques
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College of Engineering and ComputingUniversity of South Carolina
Cerebral Amyloid Angiopathy
• Characterized by deposition of A in the cerebrovasculature– Primarily A1-40
• Identified in over 90% of AD patients• A mutations: Dutch, Italian, Iowa, and
Arctic– Dutch and Italian mutations result exclusively in CAA
• Associated with an increased risk of hemorrhagic stroke
• Correlates with disease progression
Washington University ADRC
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College of Engineering and ComputingUniversity of South Carolina
A and Endothelium
Cellular degeneration
Compromised BBB integrity
Increased vessel-associated monocyte cells
ToxicityIncreased permeabilityIncreased monocyte
adhesion, transmigration
Upregulated adhesion molecule expression
Washington University ADRC
Cerebrovasculature Cell Culture
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College of Engineering and ComputingUniversity of South Carolina
Research Questions…
What form of A interacts with endothelial cells to promote activation
What signaling pathways are involved
monomer
nucleus
soluble intermediate
s
fibril
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College of Engineering and ComputingUniversity of South Carolina
Isolation of A Species
Mixture
150 mMNaCl
2-10 mMNaCl
Fibrils
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College of Engineering and ComputingUniversity of South Carolina
A model endothelium exhibitingblood-brain barrier properties was developed
Gonzalez-Velasquez and Moss (2008) J Neurochem, 104: 500-13.
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College of Engineering and ComputingUniversity of South Carolina
Endothelial Cell Assays
Adhesion Transmigration Permeability
permeabilitycoefficient
Pe
%transmigrated
cells
%adherent
cells
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College of Engineering and ComputingUniversity of South Carolina
Soluble A aggregates selectively activate endothelial cells
MIX MON SOL FIB
Gonzalez-Velasquez and Moss (2008) J Neurochem, 107: 466-77;Gonzalez-Velasquez, Kotarek, and Moss (2008) J Neurochem, 104: 500-14.
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College of Engineering and ComputingUniversity of South Carolina
Endothelial activation ismore pronouncedin the presence of smaller soluble Aaggregationintermediates
Gonzalez-Velasquez and Moss (2008) J Neurochem, 107: 466-77;Gonzalez-Velasquez, Kotarek, and Moss (2008) J Neurochem, 104: 500-14.
Adhesion Permeability
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College of Engineering and ComputingUniversity of South Carolina
NF-B and A
• NF-B is involved in the activation of endothelium – Adhesion molecule
upregulation
– Permeability
• A is capable of activating NF-B– Neuronal cells
– Glial cells
• A can activate NF-B via binding of several receptors– Receptor for advanced
glycation end products (RAGE)
– Toll-like receptors (TLRs)Hayden and Ghosh (2008) Cell,
132:344-62.
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College of Engineering and ComputingUniversity of South Carolina
CONT TNF
MON SOL
MIX
FIB
NF-B nuclear translocation is selectively stimulated by soluble A aggregates
Gonzalez-Velasquez, Reed, et al, and Moss (2010) Curr Alzheimer Res, 8: 81-94.
.
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College of Engineering and ComputingUniversity of South Carolina
NF-B activation is observed at physiological concentrations of A
Gonzalez-Velasquez, Reed, et al, and Moss (2011) Curr Alzheimer Res, 8: 81-94.
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College of Engineering and ComputingUniversity of South Carolina
CONT MON MIX FIB0
10
20
30
40
***
% A
dhes
ion
CONT MON MIX FIB0
10
20
30
40
***
% T
rans
mig
rati
on
Inhibiting NF-B nucleartranslocation abrogatesA-induced adhesion,transmigration, andpermeability
Control MG 132
Gonzalez-Velasquez, Reed, et al, and Moss (2011) Curr Alzheimer Res, 8: 81-94.
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College of Engineering and ComputingUniversity of South Carolina
Summary
• A is capable of activating endothelial monolayers– Monocyte adhesion– Monocyte transmigration– Permeability
• Specifically, soluble A aggregation intermediates are responsible for endothelial activation
• Smaller soluble aggregation intermediates are the most potent activators of the endothelium
• NF-B signaling is involved in A activation of endothelial monolayers
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College of Engineering and ComputingUniversity of South Carolina
Research AssociateFrancisco Gonzalez-Velasquez
Undergraduate StudentsCharlotte CooperTim DavisAnthony EgalMcCall RogersEmily Matherly
Graduate StudentsAdriana Reyes BarceloJoseph KotarekWill ReedDeborah Soto-OrtegaChen SuoJui-Heng TsengKelly Wilson
AcknowledgementsNational Science Foundation
CAREER Award (BES 0644826)
American Heart Association, Mid-Atlantic AffiliateBeginning Grant-In-Aid (0565387U)
Alzheimer’s AssociationNew Investigator Research Grant (NIRG-07-60412)
USC Complementary and Alternative Medicine Center
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Activation of endothelium in Alzheimer’s brain involves soluble aggregates of the amyloid- protein
F.J. Gonzalez-Velasquez, J.W. Reed,J.W. Fuseler, J.A. Kotarek,
D.D. Soto-Ortega, and M.A. Moss
University of South Carolina
College of Engineering and ComputingUniversity of South Carolina