1. THE ADDITION OF BETA-SITE
AMYLOID PRECURSOR PROTEIN
CLEAVING ENZYME INHIBITORS TO
SLOW DOWN THE CREATION OF
BRAIN PLAQUES IN MICE WITH THE
ALZHEIMER’S DISEASE
Victor Esparza
Tayo Omoniyi
P.J. Rahimi
Alexis Sleeper
Marvin Thomas, Jr.
2. There are about 5.2 million people in the
United States with the Alzheimer’s
Disease, which includes about 5 million
over the age of 65 and approximately
200,000 people under the age of 65. This
number is expected to increase to about
16 million by the year 2050.
Background
3. Alzheimer’s Disease is a degenerative
brain disease that occurs in people
usually over the age of 65. This disease is
caused by plaques in the brain that
makes the communication between
nerve cells difficult. This is the reason why
people with Alzheimer’s start forgetting
things.
Alzheimer’s Disease
4. Plaques in the brain are created by a cluster of
protein pieces. This protein is called Amyloid-beta
(sometimes called Beta-amyloid), which is
created when the BACE is cleaved. These protein
pieces clump together and block cell-to-cell
signaling at synapses. The blocking of signaling
leads to a slow, but constant and consistent cell
death which starts affecting memory and
shrinking the brain, and eventually leads to death.
Alzheimer’s in Depth
7. References
& Questions
Explanation of how we plan to conduct our
project
Results, Expected Outcomes, and Potential
Problems and Remedies are explained
Our Project
Explanation of the project
Explanation of our resources used, timeline,
and budget
A listing of our references used and a Q&A
with us
Objective
Methods
Results and
E.O.
Resources
8. The objective of our project is to add
inhibitors to limit the production of the
Beta-site APP-Cleaving Enzyme (BACE),
the enzyme that creates the Amyloid-
beta protein which, in an amount that is
vast compared to a normal person,
creates the plaques in the brain that is
associated with the Alzheimer’s Disease.
Objective
9. We will have four main groups of mice
split into four subgroups each with 25
mice for a total of 100 wild-type mice
and 300 transgenic mice. The transgenic
mice are genetically altered to have the
Alzheimer’s Disease.
Methods
10. Mice weigh around 300 milligrams and the max
dosage of a substance you can inject into a
mouse is 1/30th of their body weight, which is 10
mg. In order to test its maximum effectiveness,
we will administer different dosages of the
inhibitor starting with a 2 mg dosage, then a 5
mg dosage, and the maximum 10 mg dosage of
the Beta-secretase inhibitor.
Methods Cont.
11. Group 1 will be our control mice which do not have
Alzheimer’s Disease. Subgroup A, consisting of 25 mice, will
not receive a dosage of the Beta-secretase inhibitor.
Subgroup B will receive the minimum of 2 mg dosage.
Subgroup C will receive the 5 mg dosage and then
subgroup D will receive the 10 mg dosage. Giving the
inhibitor to the control group allows us to see its effects on
normal mice. In theory, the normal mice should also have a
decrease in Beta-secretase production, therefore
confirming the legitimacy of the decrease of beta-amyloid
in mice with Alzheimer’s disease.
Group 1 of Mice (Control/Wild-Type)
12. The second group of mice will be divided up much like the
control group. Group 2 will consist of 100 mice split into the 4
subgroups of 25 mice. These mice will have been mutated with
the Alzheimer’s disease and will be tested four months after they
have been given the disease. Since it takes around six months for
the mice to form plaque on the neurons, we will be testing group
2 to see whether or not the inhibitor can be used as a preventive
measure during the onset of the disease to keep the plaques from
forming, therefore saving neurons. Similar to group 1, subgroup A
will be given no Beta-secretase inhibitor, subgroup B will receive
the 2 mg dosage, subgroup C will receive the 5 mg dosage, and
subgroup D will receive the full 10 mg dosage.
Group 2
13. The third and fourth groups of mice will follow the same
patterns as before. These mice will be mutated with
Alzheimer’s Disease and will be tested eight and
twelve months, respectively, after mutation. These
mice should show already formed plaques of Amyloid-
beta on their neurons and will have an excess of Beta-
secretase in their brain. These two group will be used
to show if the inhibitor can be used to slow down the
progression of Alzheimer’s in mice whose progression is
further along than the other.
Groups 3 and 4
14. In order to test the effectiveness of the Beta-
secretase inhibitor, we have devised a series of
tests that target the effects of the Beta-amyloid
plaques. We will be testing their
learning/memory abilities by administering the
Morris Water Maze Test, cerebrospinal fluid for
Amyloid-beta levels, and motor coordination.
There will be a five day trial of all the tests two
weeks before and after the injection.
Testing the Effectiveness of the
Inhibitor
15. We expect the Amyloid-beta to decrease, but not fully go away. Mice that
initially have plaques are likely to not develop a further aggregation of these
plaques. Administration of the compound on mice that are predisposed to
developing plaques, but have not yet developed them, are likely to never
develop plaques or have a significantly small amount of buildup. In the
learning and memory tests, we expect mice who have not yet developed
the plaques to perform nearly as well as the control mice and the mice who
have developed the plaques to perform better than they did in the initial
trials prior to injection. The mice’s cerebrospinal fluid tests should also show a
decrease in the amount of Amyloid-beta potency in the second round of
testing as plaques should be forming at a slower rate or, even better, not
forming whatsoever. We also can expect improved motor coordination if the
inhibitor indeed decreases the protein production in the brain. Overall, the
Beta-secretase inhibitor should improve cognitive function since less neurons
are dying and the amount of plaques are decreasing.
Results
16. We expect to find a significant decrease in BACE without
completely eliminating it. The plaque in the brains of the
mice should reduce significantly as a result. Overall, the
mice who had plaque before administering the inhibitors
should have improved motor control and memory once the
inhibitor is administered. We would expect them to show
improved results during the second trials of tests. We do not
expect the transgenic mice to perform as well as the wild-
type mice during the tests, but we expect them to perform
close to the wild-type mice after administering the inhibitor.
Expected Outcomes
17. Studies show that the deletion of all the Beta-secretase to
stop Amyloid-beta production had only a minor impact on
the mice. This shows that eventually, it will lead to a
problem. Only a partial reduction of production of Amyloid-
beta should be taken to allow for better results. Reports
demonstrate that just a partial reduction and a little less
than fifty percent of the brain plaque have showed
significant improvements with the mice. These reports
conclude that these symptoms can be improved in spite of
the leftover Amyloid-beta production.
Potential Problems
18. Resources, Timeline, Budget
Budget
• Lab: $100,000
• Mice: $7,200
• Mice Necessities: $20,000
• Equipment: $150,000
• Employees ($30,000/employee): $300,000
• BACE Inhibitor: $680,000
• Miscellaneous: $17,800
Total: $1,275,000
• It will take about 15 months to complete experiment and
gather data.
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References
