Targeting cerebrovascular endothelial cells as a therapeutic approach for amyloid pathogenesis
Project Summary
The accumulation of amyloid-¿ (Aß) in the brain blood vessels can result in the development of cerebral
amyloid angiopathy (CAA). CAA is a pathological feature present concomitantly with Alzheimer’s disease (AD)
at a high frequency, highlighting a potentially important role for vascular Aß in dementias such as AD. While
the basis by which Aß mediates deleterious effect on the blood-brain barrier (BBB) is likely multifactorial,
numerous studies indicate a role for Aß mediated increases in endothelial cell permeability. The exact causes
for BBB dysfunction in CAA are not well known, however impaired clearance of Aß from the brain across the
BBB as well as a reduction in the efficacy of the perivascular drainage of A¿ have been proposed to enhance
accumulation of cerebrovascular and parenchymal amyloid deposits in the elderly. Despite our understanding
of the pathways responsible for BBB dysfunction and clearance of Aß, the availability of drugs to treat Aß
pathogenesis related disorders, CAA and AD, remains lacking. The long-term goal of this project is to develop
therapeutics that target the BBB to restore its function and maximize clearance of Aß from the brain, which is
important to prevent or delay onset of CAA and AD. The overall goal of this project is to fully characterize an
experimental endothelial BBB model as an effective high-throughput screening (HTS) format to identify
therapeutics for vascular Aß pathogenesis disorders (AD, CAA and vascular dementia). The central hypothesis
is that high-throughput screening (HTS) utilizing a highly novel cerebrovascular endothelial BBB model can be
used to identify small molecules which beneficially regulate A¿ clearance and reduce A¿ mediated increases in
BBB permeability. We will test this hypothesis by pursuing the following specific aims: 1) Utilization of a cell
line-based BBB model to screen for modulators of A¿ mediated disturbances of endothelial cell function. This
aim will be accomplished by investigating the following sub-aims: 1A) screen for compounds, in the presence
of A¿42 oligomers, for their effect on the gross permeability of cerebrovascular endothelial cells using Lucifer
Yellow (LY) as a gross permeability marker. Compounds which reduce A¿-mediated permeability will be
advanced to Sub-Aim 1B; 1B) identify stimulators of A¿ clearance across the BBB model using the gold
standard measure of iodinated A¿ as the endpoint for A¿ clearance. Hit compounds will be further examined in
Aim 2. 2) Validation and mechanistic investigation of hit compounds from Aim1 for their ability to modulate
expression of tight junction and A¿ clearance proteins (transport and degradation). This aim will be
accomplished by testing the following sub-aims: 2A) conduct secondary confirmation and establish profiles for
hits identified in Aim 1, 2B) comparison of hits kinetics for amelioration of A¿42 oligomers induced permeability
and reduced A¿ clearance in primary cerebrovascular endothelial cells with those of Sub-Aim 2A, 2C)
mechanistic investigation for improved BBB tightness, integrity and A¿ clearance by hit compounds selected
from 2B. 3) Test ability of the top 2 hits from Aim 2 to in vivo modulate vascular and parenchymal A¿
accumulation, and BBB integrity in a mouse model of CAA. Methods and technique to be used to accomplish
the above aims include in vitro cell culture, high-throughput screening, transport, permeability and clearance
studies, A¿ kinetics, microvessels isolation from brains of wild type mice, and in vivo studies in CAA model.
The data produced will provide candidate therapeutic molecules to test in future clinical studies.
