(157f) Mechanistic Inhibition of Alzheimer's-Associated Aβ Aggregation by Gold Nanoparticles

Wilson, K. A., University of South Carolina
Mahtab, R., SC State University
Jackson, K., SC State University
Moss, M. A., University of South Carolina

Alzheimer's disease (AD) is currently
the most common type of dementia and the 6th leading cause of death
in the United States.  One pathological hallmark of AD is amyloid plaques,
which deposit around the neurons in the brain.  These plaques are composed
primarily of amyloid-beta (Aβ), which is a 40-42 amino acid protein that
forms from the cleavage of the amyloid precursor protein (APP).  Aβ is a
naturally occurring protein in the body; however it self-assembles to create
aggregated structures and ultimately insoluble fibrils in a process that is
hypothesized to be closely tied to disease progression.  During self-assembly,
monomeric Aβ forms nuclei, which progress to form soluble aggregates.
These soluble aggregates can then associate or elongate to produce insoluble
fibrils.  In association, soluble aggregates laterally bind to one another to
form fibrils of an increased diameter. In elongation, monomeric Aβ binds
to the ends of the soluble aggregates to form fibrils with an increased
length.  Inhibition of Aβ aggregation has emerged as a therapeutic
strategy for AD. We have examined gold nanoparticles for their ability to halt
specific Aβ aggregation mechanisms.  Nanoparticles are of particular
interest because they cross the blood-brain barrier and have been used for drug
delivery. By employing TEM images to evaluate aggregate morphology as well as
assays that isolate Aβ aggregation mechanisms, we have tested the
mechanistic-specific inhibitory capabilities of gold nanospheres with citrate,
cetrimonium bromide (CTAB), poly acrylic acid (PAA), and poly allylamine
hydrochloride (PAH) overcoatings.

The effect of gold nanoparticles on Aβ1-40
monomer aggregation was first evaluated.  Subsequently, mechanistic-specific
assays were employed to assess the effect of gold nanoparticles on soluble Aβ1‑40
aggregate elongation and association.  These assays utilized thioflavin T to
monitor increases in amyloid material, dynamic light scattering to monitor
increase in aggregate size, and TEM to observe changes in aggregate morphology.

Gold nanoparticles overcoated with
citrate, CTAB, and PAH were observed to have little effect on the quantity of
amyloid material formed during Aβ1-40 monomer aggregation. TEM
images (Figure 1), however, demonstrate that aggregations performed in the
presence of CTAB overcoated nanoparticles favored aggregate association over
elongation.  Mechanistic-specific assays confirm these trends and directly tie
aggregation mechanisms to aggregate morphology.  PAA nanoparticles abrogated
the formation of amyloid material in monomer aggregation assays.  Furthermore,
this inhibition was observed at a substiochiomeric ratio of nanoparticles to Aβ
of 1:200.  Elongation and association assays further define the mechanism of
this inhibition.

Together these results demonstrate that
gold nanoparticles serve as effective inhibitors of Aβ self-assembly that
modify aggregation at substiochiometric concentrations. Furthermore, inhibition
of Aβ aggregation by gold nanoparticles is mechanistic-specific in a
manner that is dependent upon the nanoparticle coating. Determining the
mechanisms by which these various nanoparticles inhibit Aβ self-assembly
may be used to develop an inhibitor capable of targeting multiple mechanisms.



Figure 1: TEM
image (100,000X), 40µM
monomer aggregated in the presence of
0.2nM CTAB nanoparticles.  Presence of the CTAB nanoparticles allows for the
development of associated and non-elongated aggregates.