Facing Alzheimer’s ​0

Facing Alzheimer’s: What You Don’t Know Can Kill You

Olivia Wang

Mrs. Ireland

Intern/Mentor

March 6, 2019
Facing Alzheimer’s 1

Abstract

Currently ranked as the sixth leading cause of death in the United States alone,

Alzheimer’s disease is a serious condition that arises in someone in the United States every 65

seconds. Roughly 10 percent of adults are diagnosed Alzheimer’s after the age of 65 and at least

32 percent after the age of 80. More so, 25 percent of those originally diagnosed with

Alzheimer’s are misdiagnosed, commonly due to the lack of a full diagnostic report and series of

tests​[1]​. The misdiagnosis is due in full to the insufficiency of appropriate testing for patients.

Alzheimer’s is a heavily genetically influenced degenerative disease and the need for companies

to create a new drug or treatment plan for patients with Alzheimer’s. There are three main

reasons why there had been no successful advancements in the last decade and a half in the race

for finding an Alzheimer’s drug cure. The first is because of the complexity of the disease itself.

The symptoms of Alzheimer’s do not actually begin to behaviorally become prevalent until there

has been irreversible damage done to the brain. The second reason is because there are no good

forms of diagnosis for us to really tell at an early stage, if the patient biologically has

Alzheimer’s disease. Yes, there are tests that show if you have Alzheimer’s disease, but a

majority of Alzheimer’s patients don’t actually get this test. The two current existing tests for

Alzheimer’s are PET scans and spinal taps. In other words, a $4000 scan that gives a really high

definition picture for what is going on in a patient’s brain, and an extremely painful process

which includes sticking a needle in a patient’s spine to obtain cerebrospinal fluid. Many of these

patients with Alzheimer’s disease are in their 60’s and 70’s. Using these two forms of testing to

diagnose a patient with Alzheimer’s disease is completely unreasonable because they are costly

and invasive. It is the very reason for so many cases of misdiagnosed Alzheimer’s. The third is

because it is so hard to find good candidates for clinical trials. Patients who are misdiagnosed
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with Alzheimer’s are participating in these clinical trials and are, as a result, skewing the data

substantially. Scientists currently have no way to study Alzheimer’s patients for long enough to

test memory-loss prevention drugs. It was so critical to the point that in 2012, former President

Barack Obama created a national plan called the National Alzheimer’s Act in hopes to find a

successful treatment plan or cure for Alzheimer’s disease by 2025. We are 7 years into this plan

and we haven’t had much substantial progress in this race.

Introduction

A​lzheimer’s disease (AD) is a progressive neurodegenerative disease, which means that it

is the gradual decay of a person’s brain associated with cognitive decline. The decline affects a

person’s cognitive abilities like being able to think, remember, reason,​ and behavioral abilities

like being able to walk, talk, go to the bathroom, etc.. The dementia symptoms in Alzheimer’s

gradually worsen over a number of years. In its early stages, memory loss is mild, but with

late-stage Alzheimer's, individuals lose the ability to respond to their environment. Alzheimer’s

disease is heavily affected by genetics. With Alzheimer’s, there is one main genetic variant

linked to the regression. This is the APOE gene. The APOE gene is responsible for the creation

of the class of proteins called the Apolipoprotein. The Apolipoproteins produce proteins that

cause the degeneration in the brain.There are many different variants of this gene. The e4 allele

of APOE gene increases the risk of Alzheimer’s. People who inherit one copy of the the allele

already have an increased risk; people who inherit two copies have an even greater risk. The

APOE gene provides instructions for making a protein called apolipoprotein E. Apolipoproteins

are partially responsible for the pathological occurrences of Alzheimer’s. Biologically,

Alzheimer’s occurs when there is an accumulation of one protein called beta-amyloid and
Facing Alzheimer’s 3

another protein called tau. When tau accumulates into neurofibrillary tau tangles and

beta-amyloid accumulates into plaques, severed neurodegeneration occurs. This accumulation of

apolipoproteins precisely shows the severity of the disease in a patient. Targeting the amount of

Apolipoprotein from a simple blood sample from a patient to create a new diagnostic test can

show the state of the Alzheimer’s in its degeneration, can be detected by a series of

electrophoresis tests on the Alzheimer’s cell line, and can ease the vexatious uneasiness of those

who are unaware of the progression Alzheimer’s disease in a patient. A full diagnosis of

Alzheimer’s disease is necessary for doctors working on clinical trials, for patients unsure of the

state of their disease, and for pharmaceutical companies looking for treatments and can be

achieved by targeting the creation of a test that analyzes a patient’s blood.

Background Information

Alzheimer’s disease (AD) is a progressive neurodegenerative disease that is associated with

cognitive decline. It is also the most common form of dementia in the elderly.​ Alzheimer’s

disease is a form of dementia, which is the loss of cognitive function - thinking, remembering,

and reasoning - and behavioral abilities to such an extent that it interferes with a person’s daily

life and activities​[14]​. Alzheimer's is a progressive disease, where dementia symptoms gradually

worsen over a number of years. In its early stages, memory loss is mild, but with late-stage

Alzheimer's, individuals lose the ability to carry on a conversation and respond to their

environment​[5]​. It is the most common cause of dementia in older adults. Alzheimer’s Disease is

a neurodegenerative disease that is heavily affected by genetics. There have been studies that

have looked into family studies and twin studies to further the knowledge in how the biology of

the genetics works. It has been discovered that the there are specific gene mutations that link
Facing Alzheimer’s 4

directly to Alzheimer’s disease​[13]​. Alzheimer’s disease occurs when there is an accumulation

of beta-amyloid or neurofibrillary tau tangles in the brain. Alzheimer's has no current cure, but

treatments for symptoms are available as the research for a cure continues. However, increasing

efforts are coming into play in order to carve a new path to finding treatment plans for patients.

Apolipoprotein role in Alzheimer’s

The abundance of Apolipoproteins can be measured from the patients blood and shows a direct

correlation in the progression of Alzheimer’s. From a blood sample, the genetic variant tied to

Alzheimer’s in a patient can be detected. Alzheimer’s disease is a heavily genetically influenced

disease. In an Alzheimer’s patient, a specific gene is linked to the influence of onset of

Alzheimer’s. The gene that is linked to Alzheimer’s lies within the chromosomes 1, 19, and 21.

The one main genetic variant linked to the regression is the APOE gene. The APOE gene is

responsible for the creation of the class of proteins called the Apolipoprotein E4. The

Apolipoproteins produce proteins that cause the degeneration in the brain​[15]​.

Effects of Alzheimer’s Disease on Patients

Alzheimer’s disease is an illness of the brain. It causes large numbers of nerve cells in the brain

to die. This affects a person’s ability to remember things, think clearly, and use good judgment.

People with Alzheimer’s disease have trouble doing everyday things like driving a car, cooking a

meal, or paying bills. They may get lost easily and find even simple things confusing. Some

people become worried, angry, or violent​[16]​. Human apolipoprotein E (ApoE) is associated

with high cholesterol levels, coronary artery disease, and mostly in Alzheimer’s disease. There

are different stages of Alzheimer’s disease. In the early stage of Alzheimer's, most people
Facing Alzheimer’s 5

function independently. They may still drive, take part in social activities, work and even athletic

events. The early stage of Alzheimer’s may last for many years. The middle stages of

Alzheimer's are typically the longest and can last for many years. As dementia progresses, the

person with Alzheimer's will require a greater level of care. During the middle stages of

Alzheimer's, damage to the brain can make it difficult to express thoughts and perform routine

tasks. The person with Alzheimer's may begin jumbling words, having trouble dressing, getting

frustrated or angry, or acting in unexpected ways, such as refusing to bathe. The late stage of

Alzheimer's disease may last from several weeks to several years. As the disease progresses,

intensive, around-the-clock care is usually required. As the disease advances, the needs of the

person living with Alzheimer's will change and deepen. A person with late-stage Alzheimer's

usually: have difficulty swallowing and eating, need assistance in walking, and may eventually

be unable to walk, need full time assistance in personal care, and is vulnerable to infections.​[2]

Effects of Apolipoproteins in Alzheimer’s Disease

The Apolipoprotein gene variant is very important and a crucial part in Alzheimer’s research.

This gene variant only determines if the patient is more susceptible to getting Alzheimer’s

disease. However, the amount of the beta-amyloid, the protein that causes Alzheimer’s, is what

determines how far in the progression of Alzheimer’s a patient is​[3]​. ​Genome-wide association

studies have confirmed that the E4 allele of ​APOE​ is the strongest genetic risk factor for AD.

The presence of this allele is associated with increased risk for both early-onset AD. ​The

amyloid precursor protein gene is linked to the chromosomes 1 and 19. The targeted protein,

Apolipoprotein E gene variant, has been reported to be located on the chromosome 19 for

patients with late-onset Alzheimer’s. In addition, it has also been discovered that the
Facing Alzheimer’s 6

chromosome 21, the chromosome known for its association with Trisomy 21, or Down

Syndrome, is a candidate for Alzheimer’s genes. Those with Down Syndrome have a higher risk

and link into getting Alzheimer’s disease​[13]​. The Apolipoprotein E(APOE) is a major

cholesterol carrier that supports lipid transport and injury repair in the brain. APOE​ ​polymorphic

alleles are the main genetic determinants of A​lzheimer’s disease risk. There are two main alleles

in determining risk: the E4 allele and the E3 allele. Individuals carrying the E4 allele are at

increased risk of AD compared with those carrying the more common E3 allele. Presence of the

APOE​ E4 allele is also associated with increased risk for cerebral amyloid angiopathy and

age-related cognitive decline during normal ageing​[12]​.The apolipoprotein production associated

with the E4 allele is thought to be involved in the formation of a plaque. These plaques are

beta-amyloid plaques. Beta-amyloid is a peptide that causes neurological degeneration. It is the

hallmark of Alzheimer’s disease. The E4 allele allows for a higher production of beta-amyloid

and an increased risk in onset of Alzheimer’s.

Using An Electrophoresis Test

Because Apolipoproteins are bound together, the DNA fragments need to be separated. A Gel

electrophoresis test can separate these DNA fragments by filtering them by size and charge. An

electrophoresis test on an Alzheimer’s cell line should be able to detect Apolipoprotein

abundance without being invasive or costly. After the cell line from an Alzheimer’s patient is

isolated, the genetic variant tied to Alzheimer’s disease can be targeted and amplified. This

amplification allows for an easier detection for Apolipoproteins.

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Presence of Apolipoprotein RNA in Exosomes

The presence of Apolipoprotein RNA in exosomes can show the regression of Alzheimer’s

disease. In the brain, there are small vesicles secreted by neurons that function in intercellular

communication called exosomes. These exosomes have been observed to play a role in spreading

pathological misfolded proteins. In Alzheimer’s disease, there are two types of lesions: Amyloid

plaques and neurofibrillary tangles. The role of exosomes in the metabolism and secretion of

APP and Tau proteins and their subsequent impact on AD pathogenesis is studied in this review.

The biogenesis and function of exosomes impact amyloidogenic processing and tau pathology.

The exosomes have a “double-edged sword” effect on the progression of AD. The discovery of

the path in which exosomes travel could provide insight into the diagnosis of AD and the

platform to allow therapeutic drugs for AD​[18]​.

Experimentation With Electrophoresis and Exosomes

In an electrophoresis test, the mutated cells are isolated and filtered to get rid of extremities.

RNA strands from the specific cell line are isolated and amplified. This process is completed my

using High-Capacity cDNA Transcription. RNA is converted into cDNA by using the

technology, Reverse-Transcriptase Polymerase Chain Reaction (RT-PCR). This approach is used

to study gene expression. Although RT-PCR and the traditional PCR process both produce

multiple copies of particular DNA isolates through amplification, the applications of the two

techniques are fundamentally different. Regular PCR is used to exponentially amplify target

DNA sequences. RT-PCR is used to copy expressed genes by reverse transcribing the RNA that

was extracted into cDNA (complement DNA) through the use of reverse transcriptase.

Subsequently, the newly synthesized cDNA is amplified using traditional PCR. Therefore,
Facing Alzheimer’s 8

instead of having an amplified target DNA sequence, we can look at the amplified cDNA

sequence that was reverse transcribed from RNA​[6]​. Comparing the results from testing a

mutated cell line and a non-mutated cell line to discover the presence of apolipoproteins in the

cell line using electrophoresis tests, experimentation has proven to be successful in detecting

exosomal presence in electrophoresis.

RNA Expression of Apolipoprotein in Blood Sample

Many people know that Alzheimer’s is a hereditary disease. However, what they don’t know is

how it works and they there is a probable chance that it can even be transmissible through blood.

A study has found that Alzheimer’s protein can spread between mice that share a blood supply,

causing brain degeneration. Other diseases like Creutzfeldt-Jakob Disease (CJD) can spread

through meat products or blood transfusions infected with prion proteins because they have

misfolded proteins that are known to cause CJD. Like CJD, Alzheimer’s also has involves a

misfolded protein called beta-amyloid. There has been recent evidence that beta-amyloid may be

able to spread like the prions in CJD. A recent tried conjoining a healthy mouse and a mouse

with Alzheimer’s plaques. They surgically attached the two mice in a way that made them share

a blood system. The study proved to be truthful in the transference of those beta-amyloid

proteins from the mouse with Alzheimer’s to the healthy mouse​[10]​. RNA transcriptome in AD

can help the understanding of how the disease presents itself. The effects of white matter

hyperintensities on AD blood transcriptome has not been considered. The RNA from whole

blood was processed on whole-genome microarrays. It is suggested that blood-based biomarkers

could be useful for evaluating diagnosis, pathogenesis and progression of AD. RNA expression

in blood of patients with AD has been assessed in several recent independent studies.. The aim of
Facing Alzheimer’s 9

this study was to examine RNA expression in blood of AD patients with and without WMH as

compared to controls.The study shows that a number of the genes regulated in this study have

been reported to be regulated in other studies of both AD blood and AD brain, and strongly

support a systemic effect of AD that includes altered RNA expression of peripheral immune

cells. Further validation in an independent cohort to specifically test the sensitivity and

specificity of the identified genes to predict AD is needed​[4]​.

Purpose For Finding An Alternative Diagnostic Test

Finding an alternative form to testing for Apolipoprotein abundance in patients is necessary

because it allows for more effective treatment procedures. For the 113 years since Alzheimer’s

disease was discovered, neither a treatment nor a cure has been found. This absence of a

treatment for Alzheimer’s has many negatory costs.

Benefits for Patients

Sandra Day O'Connor, the first woman on the Supreme Court, announced Tuesday in a frank and

personal letter that she has been diagnosed with "the beginning stages of dementia, probably

Alzheimer's disease."O'Connor said doctors diagnosed her some time ago and that as her

condition has progressed she is "no longer able to participate in public life." After her 2006

retirement from the high court O'Connor had appeared around the country championing an

educational organization she founded and serving as a visiting appeals court judge, among other

activities. But she stopped speaking publicly more than two years ago. Her Alzheimer’s was not

diagnosed until she began showing symptoms of dementia​[17]​. However, finding a new

diagnostic test does not just benefit people like Sandra. Since Bill Gates announced his
Facing Alzheimer’s 10

investment in research on Alzheimer’s disease, he received widespread appreciation and support.

When asked on the reason behind his newfound support, Gates revealed that it was because his

father, Bill Gates Sr. suffered from Alzheimer’s disease. Gates further explains that the reason

with no cure found for Alzheimer’s yet is because it is a chicken-and-egg issue. Without an

accurate, precise, painless, and non-costly form of diagnosis, Alzheimer’s will be unable to be

treated or cured. Because of this, Gates invests more than $30 million towards the project,

Diagnostics Accelerator, of the Alzheimer’s Drug Discovery Foundation​[8]​. Finding an

alternative testing allows for closure from patients. At age 49, a woman by the name of Julie

Gregory paid an online service to sequence her genes. She was not originally looking for it, but

soon found out that she had two copies of the ApoE4 gene. This gene is strongly linked to

Alzheimer’s. Many scientists say that there is no other way to detect whether you have

dangerous plaques that are known to cause Alzheimer’s unless you get a $4,000 PET scan or an

extremely painful spinal tap. While genetic tests can help predict risk of getting the disease, it is

still unable to reveal the state of the disease in your brain. Many people will go years without

ever showing a single sign of Alzheimer’s even if they had it. Another woman, acknowledged as

D., also found out about her ApoE genotype. After finding out, she, like many upon finding out,

began to have the looming thought/ question about when and if she would be getting

Alzheimer’s. The biggest fear with knowing that someone carries the ApoE4 gene variant is that

those people would never know what state the disease is at in their head, when they would begin

showing symptoms of the disease, and how bad the disease can get. The big issue is that people

in the early stages of Alzheimer’s will never know unless they got tested​[11]​.
Facing Alzheimer’s 11

Benefits for Researchers and Pharmaceutical Companies

Alzheimer’s disease is expected to increase prevalently in the near future. As of now, there is no

definitive diagnosis process that can determine the state of the Alzheimer’s progression in a

patient with ease in accuracy and precision. Many patients are incorrectly diagnosed.

Cerebrospinal Fluid biomarkers have been discovered to be the closest thing to getting an

accurate diagnosis. However, just like many things on this earth, CSF has its flaws and is

difficult to obtain. Although these CSF biomarkers have supported the identification of tau

tangles in the brain, the performance in giving an appropriate diagnosis is still at risk because it

only gives a moderate diagnosis. It is also possible that when running labs on the CSF, it was

frozen throughout the duration in order to preserve the biomarkers, which could have caused

error in the process​[9]​. Out of all dementia cases, roughly 6o to 80 percent of these cases are

Alzheimer’s disease. The number of cases introduced each year is ever increasing. Nearly every

66 seconds, there is a new case of Alzheimer’s introduced. There are fairly effective psychiatric

medication options for the anxiety that comes with living with constant confusion. There are

three main reasons that scientists have not discovered a drug to treat Alzheimer’s disease, among

them being that there is 25% chance that the patients in clinical trials where these drugs are

tested have, in fact, been misdiagnosed with Alzheimer’s​[7]​.

Conclusion

In the last 113 years, since Alzheimer’s disease was discovered, no cure or treatment plan has

been procured. An alternative to accurately and precisely diagnosing Alzheimer’s disease is a

necessary advancement in medicine because it affects a large portion of the current population.

The amount of Apolipoproteins in a cell line from a patient can help to determine the presence of
Facing Alzheimer’s 12

the Alzheimer’s and its position in the pathway to degeneration of a patient’s brain, as well as,

help patients gain closure for their disease, even if there is not cure discoverable. Although

current Alzheimer's treatments cannot stop Alzheimer's from progressing, they can temporarily

slow the worsening of dementia symptoms and improve quality of life for those with Alzheimer's

and their caregivers. Today, there is a worldwide effort under way to find better ways to treat the

disease, delay its onset, and prevent it from developing.

Research Methods and Data Collection

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Works Cited

 Alzheimer’s Association. “Facts and Figures.” ​Alzheimer's Disease Facts and Figures​,
Alzheimer's Association[1]

 Alzheimer’s Association. “Stages and Behaviors.” ​Alzheimer's Disease and Dementia​,

Alzheimer's Association, www.alz.org/help-support/caregiving/stages-behaviors.[2]

 Apostolopoulou, Maria. Personal Interview. September 18, 2018 [3]

 Bai, Z., Stamova, B., Xu, H., Ander, B. P., Wang, J., Jickling, G. C., . . . Sharp, F. R. ​Distinctive
RNA Expression Profiles in Blood Associated With Alzheimer’s Disease After Accounting
for White Matter Hyperintensities​ (2014, July 1). [4]

 Documentary, Best. ​Alzheimers - One of the Greatest Medical Mysteries of Our Time,​ YouTube,
30 Nov. 2016. [5]

 Farrell, Robert E. ​RNA Methodologies: a Laboratory Guide for Isolation and Characterization​.
Academic Press, an Imprint of Elsevier, 2017. [6]

 
Foley, Katherine Ellen, and Katherine Ellen Foley. “Why the Pharmaceutical Industry Is Giving
up the Search for an Alzheimer's Cure.” ​Quartz,​ Quartz, 28 Jan. 2019. [7]

 Gates, Bill. “Why Diagnosing Alzheimer's Today Is so Difficult-and How We Can Do Better.”
Bio | Bill Gates,​ Gatesnotes, 17 July 2018. [8]

 Genius, J, Klafki, H, Benninghoff, J, Esselmann, H, Wiltfang, J. ​Current Application of

Neurochemical Biomarkers in the Prediction and Differential Diagnosis of Alzheimer’s
Disease and Other Neurodegenerative Dementias. S ​ pringer-Verlag, 2012. [9]

 Hamzelou, Jessica. “Alzheimer's May Be Able to Spread through Blood Transfusions.” ​New
Scientist​. [10]

 Kennedy, Pagan. “What If You Knew Alzheimer's Was Coming for You?” ​The New York Times,​
The New York Times, 17 Nov. 2017. [11]

 
Liu, Chia-Chen, et al. “Apolipoprotein E and Alzheimer Disease: Risk, Mechanisms, and
Therapy.” ​National Institute of Aging​, National Institute of Health , 29 July 2013.[12]
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 Matsuyama, Steven S., Kumar, Vinod, and Carl Eisdorfer. ​Advances in the Diagnosis and
Treatment of Alzheimer’s Disease: Chapter 4: Genetics of Alzheimer’s Disease.​ Springer,
1998. [13]

 National Institute of Aging. “Alzheimer's Disease Fact Sheet.” ​National Institute on Health,​ U.S.
Department of Health and Human Services, Accessed March 1, 2019. [14]

 National Institute of Aging. “How Alzheimer’s Changes the Brain.” ​National Institute on Aging,​
U.S. Department of Health and Human Services, 16 May 2017. [15]

 National Institute on Aging. ​Understanding Alzheimer's Disease: What You Need to Know.​ U.S.
Department of Health and Human Services, 2017. [16]

 Press, The Associated. “Sandra Day O'Connor Announces Likely Alzheimer's Diagnosis.” ​The
New York Times​, The New York Times, 23 Oct. 2018. [17]

 Xiao, T., Zhang, W., Jiao, B., Pan, C., Liu, X., & Shen, L. (2017, February 22). NIH [18]