Apolipoprotein Role in Alzheimer’s And Effects of Alzheimer’s Disease on Patients

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Introduction

A little over a year ago, there was a sweet 90 year old woman at a nursing home. Her name was Analisa Caroler. During her stay at this nursing home, she used to tell the most amazing stories of her past. However, a year ago, she began showing signs of Alzheimer’s. She was shocked. 20 years ago, she discovered she carried a genetic marker for Alzheimer’s. Because no symptoms ever surfaced before she was 85, her doctors felt that her Alzheimer’s would never surface. Still, during these 20 years, she never knew if she was going to have Alzheimer’s or when she was going to begin showing symptoms. This was because she never received a thorough diagnostic test on her Alzheimer’s disease. 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. The misdiagnosis is due in full to the insufficiency of appropriate testing for patients. Alzheimer’s is a heavily genetically influenced degenerative disease. 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. However, detecting the abundance of Apolipoprotein can, currently, only be done two ways: PET scans and Spinal Taps, which are expensive and invasive, causing many patients to be dissuaded of getting a full diagnosis. 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 disease (AD) is a progressive neurodegenerative disease associated with cognitive decline and is the most common form of dementia in the elderly. Dementia 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[13]. 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[4]. 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 directly to Alzheimer’s disease[12]. Alzheimer’s has no current cure, but treatments for symptoms are available and research continues. However, 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. In an Alzheimer’s patient, the gene that is linked to Alzheimer’s lies within their genes. From a blood sample, the genetic variant tied to Alzheimer’s in a patient can be detected. Apolipoprotein E is a major cholesterol carrier that supports lipid transport and injury repair in the brain. APOE polymorphic alleles are the main genetic determinants of Alzheimer’s disease risk: 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[11].

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[15]. Alzheimer’s disease occurs when there is an accumulation of beta-amyloid or neurofibrillary tau tangles in the brain. AD is a heavily genetically influenced disease. 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 E4. The Apolipoproteins produce proteins that cause the degeneration in the brain[14]. Human apolipoprotein E (ApoE) is associated with high cholesterol levels, coronary artery disease, and mostly in Alzheimer’s disease.

Effects of Apolipoproteins in Alzheimer’s Disease

The Apolipoprotein gene variant is very important an crucial 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[2]. Genome-wide association studies have confirmed that the ε4 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 chromosome 21, the chromosome known for its association with Down Syndrome, is a candidate for Alzheimer’s genes. Those with Down Syndrome have a higher risk and link into getting Alzheimer’s disease[12].

Using An Electrophoresis Test

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 easy detection.

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[17].

Experimentation With Electrophoresis and Exosomes

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, instead of having an amplified target DNA sequence, we can look at the amplified cDNA sequence that was reverse transcribed from RNA[5]. 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[9]. 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 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[3].

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[16]. However, finding a new diagnostic test does not just benefit people like Sandra. Since Bill Gates announced his 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[7]. 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[10].

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[8]. 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[6].

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 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.

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