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The role of CRISPR-Cas9 for treating Human immunodeficiency virus and Alzheimer's

disease

Human immunodeficiency virus (HIV) infection and Alzheimer's disease (AD) are two

terrible diseases with major unmet clinical requirements. AD is a neurological disease that

gradually impairs memory and cognitive function, causing significant challenges for both the

affected person and their caretakers. Extensive research has been conducted, yet conclusive

causes and practical solutions are still unclear. While highly active antiretroviral therapy

(HAART) revolutionized HIV treatment, it does not eliminate the virus or undo any damage

already done. As a result, research for a cure and issues related to long-term infections must

continue (Rodriguez-Penney et al., 2013).

It is unknown exactly what causes AD, although it most likely involves a complicated

interaction between environmental & genetic variables. Some genes, such as PSEN1 and PSEN2,

contribute to familial forms, whereas others, such as APOE-e4, enhance the risk (Guerreiro et al.,

2012; Serrano-Pozo et al., 2011). The condition is characterized by disordered protein aggregates

in the brain, such as neurofibrillary tangles and amyloid plaques, which impede neuronal

connection and function and cause a gradual decline in cognitive function. HIV is a retrovirus

that attacks particular immune cells, gradually impairing the body's defenses against infections. It

multiplies and spreads by integrating its viral DNA into the host's genome and taking over

biological processes. The ongoing process of replication and immune system activation leads to

extensive harm to different organs and systems (Knopman et al., 2021).

The most noticeable early symptom of AD is usually memory loss, which develops

gradually over the years. As the illness worsens, individuals experience trouble speaking,
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thinking, making decisions, and performing daily tasks. Eventually, they become totally

dependent on other people. Around 50 million people are thought to have AD worldwide, and

predictions indicate that number will triple by 2050 (Mukadam et al., 2022). HIV infection

proceeds differently in each person due to a variety of reasons, but in the absence of therapy, it

leads to AIDS, a condition marked by opportunistic infections and significant

immunodeficiency. With HAART, HIV-positive people's life expectancy and quality of life have

increased drastically, and the incidence of AIDS-related fatalities has decreased noticeably.

However, age-related comorbidities like cardiovascular disease and neurocognitive impairment

are more common in people on HAART (Moutinho, 2022). In a nutshell there are a lot of unmet

clinical requirements related to HIV and Alzheimer's. Even though research has advanced, these

difficult and life-threatening illnesses still require effective treatments and cures.

The need for ongoing research and the creation of better treatments is driven by the

global burden of HIV and Alzheimer's disease. With roughly 850,000 people living with

dementia, mostly Alzheimer's disease, the UK confronts a serious dementia crisis. The aging

population is expected to cause a sharp increase in this number. According to projections, the

number of cases worldwide is expected to triple by 2050, affecting around 50 million people

(Tusch et al., 2022). Despite advancements, HIV continues to be an international health problem.

An estimated 38 million people worldwide were predicted to be HIV positive as of 2021, and the

number of new infections and AIDS-related casualties continues to rise at a rapid pace. Due to

the disease's disproportionate impact on particular populations, targeted treatments and better

access to healthcare are required (Presanis et al., 2021).

There is currently no known cure for Alzheimer's disease; the main goals of medications

like memantine and cholinesterase inhibitors are to control symptoms and halt the illness's
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progression. Unfortunately, some drugs have adverse effects like vomiting and vertigo.

Furthermore, research is still ongoing on further interesting treatment approaches that focus on

the root cause of illness mechanisms (Moutinho, 2022). Highly active antiretroviral treatment

(HAART) has changed the management of HIV infection. People are able to live long and

healthy lives because of this combo therapy's efficient viral suppression. Nonetheless, lifetime

devotion to HAART is essential because it is not a cure. Long-term use can also result in drug

resistance and negative effects, which emphasizes the necessity for better treatments with less

adverse reactions and possibly even therapeutic potential (Xu & Ikezu, 2008).

The novel gene editing tool CRISPR-Cas9 presents a viable way to address the unmet

medical requirements in HIV and Alzheimer's disease. The mentioned methodology is

imperative in fixing and introducing genes by precisely altering the human genetic alignment.

According to De Plano et al (2022), the accumulation of the amyloid beta plaques which is a

disordered protein aggregates particularly in Alzheimer's disease is the primary cause of

neurodegeneration. The issue of neurodegeneration can be mitigated through the application of

CRISPR-Cas9 techniques with one main tactic being elimination of the amyloid beta’s gene

producing agents such as BACE1. CRISPR-Cas9 has the capacity to effectively limit the

production of disordered protein aggregates and contain the progression of the Alzheimer's

disease through gene editing. Besides, according to a study done by Bhardwaj (2021), the author

asserts that the pre-existing amyloid beta clumps can be broken down by inserting genes

expressing enzymes through CRISPR-Cas9. Therefore, the primary course of CRISPR-Cas9 is to

completely or partially eliminate the accumulated disordered protein aggregates in the body

system. Lastly, Karimian et al (2020) argue that neuronal degeneration can be prevented by
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deploying CRISPR-Cas9 to modify genes which are fundamental for health and survival of the

vital to neuronal which are associated with improved cognitive functioning.

HIV has two key regions that are targeted by CRISPR-Cas9 for functional cure to be

affected. Firstly, the CRISPR-Cas9 targets the HIV co-receptor CCR5 and permanently disables

it to ensure that the virus does not infiltrate immune cells. In the case of inhibition of the CCR5,

the body cells become immune to the HIV infection, and hence controlled (Saayman et al.,

2015). Secondly, more emphasis is currently underway through comprehensive research on the

possibility of deploying CRISPR-Cas9 to introduce coding genes to prevent replication or

assembly of the HIV in the body of the host. According to Bhowmik & Chaubey (2022), the

"molecular shields" of immune cells might stop the virus from spreading and doing damage.

However, it is worth noting that the research on the use of CRISPR-Cas9 for HIV and

Alzheimer's disease is in early stages and more is expected to unfold with numerous obstacles.

The challenges are associated with the possibilities of CRISPR-Cas9 damaging the body immune

system and untargeted genes by modifying the unintended sections of the DNA through editing

tools. Besides, the challenges can be considered on the ethical grounds where the next

generations are viable to germline editing of the genes since they are inheritable. Despite these

challenges, CRISPR-Cas9 holds great promise for addressing the unfulfilled clinical criteria in

HIV and Alzheimer's illness. Therefore, to fully realize the success of the CRISPR-Cas9 on

mitigating of HIV and Alzheimer's illness, the research and development community should

within ethical considerations to avoid any harm to the future generation.

Figure 1: Targeting HIV-1 with CRISPR (Rohn et al., 2018)

While CRISPR-Cas9 holds enormous promise for treating HIV and Alzheimer's disease,

its application is contingent upon a number of critical factors. These include the distribution

method and specific design decisions, all of which affect the efficacy and security of the strategy.

To reach the intended cells, the delivery method should be appropriate. Besides the viral vector

being the appropriate delivery method, it is limited to the conveying gene editing and may trigger

immune responses. Contrarily, the non-viral vectors are limited to penetrate some body tissues

which make them to have reduced immunogenicity. However being invasive, injection can be

easily directed to the targeted cells, including the brain regions infected by Alzheimer. Targeting

the intended gene is fundamental in realizing full success of the CRISPR-Cas9 application (Cai

et al., 2018). The treatment of Alzheimer's, involves the targeting of amyloid beta or neuronal

survival in the related genes. On the other hand, treatment of HIV involves the introduction of

antiviral genes or CCR5 disabling. During the treatment process, it is necessary to select
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appropriate version of Cas9 to ensure that gene editing and is successful and safe. Besides, this

helps in reducing the consequences of off-target since different types of Cas9 enzymes produce

distinctive degeneration degree. RNA design can help in reducing off-target consequences by

directing CRISPR-Cas9 on targeted genes. According to Wigdahl (2014), the reduction of off-

target alteration enhances CRISPR-Cas9 specificity.

Figure 2: Targeting Alzheimer's disease with CRISPR (Gearing, 2016)

In Alzheimer's disease, crossing the blood-brain barrier is critical. Lipid nanoparticles are

promising, but direct viral vector injection may be a better way to deliver drugs to certain brain

areas. According to study done by Cai et al (2018), immunological responses and safety concerns
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must be carefully evaluated. To reach the required number of cell count in the body, it is

necessary to attain HIV systematic delivery which of importance involves the viral vectors which

target particular cells after alteration. To ensure a long term treatment optimization, it is essential

to make sure that the altered cells survive within the body immune system as long as possible.

According to Wang et al (2020), optimizing guide RNA design is critical to minimizing off-

target effects in both illnesses. The RNA designs are continuously improved through accuracy

and safety of computational forecasting and Cas9 variations techniques Advances in high-

fidelity Cas9 variations and computational forecasting techniques.

CRISPR-Cas9 treatment techniques for HIV and Alzheimer's experience various

uncertainties during the clinical trials. Such obstacles limits the usability and viability of other

related researches associated with treating then mentioned diseases. One of the main challenges

highlighted by various investigated studies surrounds getting the CRISPR-Cas9 components past

the blood-brain barrier. This is because; the blood-brain barrier is a highly selective membrane

that prevents the CRISPR-Cas9 from reaching the human brain. The nanoparticles and viral

vectors, which are the current techniques, are liable to safety and efficacy issues. Consequently,

it is important to reduce the accidental genome alterations through CRISPR-Cas9 targeting

precision. In cases where a broader application of CRISPR-Cas9 is thought through, the

challenges of off-target impacts must be taken into consideration. On the other hand, the

application of gene editing in the brain has posed ethical questions since they can be inherited by

the future generation. Therefore, according to Barman et al (2020), the healthcare fraternity must

consider ethical limitations before approving global application of CRISPR-Cas9.

The immune cells that are altered must remain active in long term to achieve successful

viral suppression besides the treatment of HIV by blocking CCR5 or adding CCR5. However, a
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more thorough research is needed to provide long-term solution to application of CRISPR-Cas9.

When an individual has more than one strain of virus in the body, it impacts the usefulness of

CRISPR-Cas9 targeting. Therefore, to successfully address the intrinsic variability in the

treatment of HIV, there is need for further research in the functionality of CRISPR-Cas9 editing

(Wigdahl, 2014). Currently, there are numerous distinctive Alzheimer's and HIV treatment

options being investigated and they present both pros and cons. Despite the techniques showing

potentials in the research studies, the progression of the diseases still remains a big challenge to

be stopped or reversed. As concluded by Ashrafian et al (2021), there are clinical trials for the

treatment of Alzheimer's through antibodies that are aimed at targeting particular molecules

associated with Alzheimer's. However, the long term efficacy and safety should be determined

before such medicines are released into the market.

The development of broadly neutralizing antibodies (bNAbs), which can neutralize a

range of HIV strains as well as drugs that target various aspects of Alzheimer's disease, such as

tau tangles and amyloid beta plaques, offers hope for an effective treatment in the fight against

HIV. According to a study done by Kumar et al (2018), the development of bNAbs that can be

effective and safe in long run is still faced with uncertainties. However, in addition to the

treatment mentioned for long term treatment option, the introduction of antiviral genes by gene

therapy which employing lentiviral vectors remains applicable. Poeschla et al (1996) asserted

that in the process of HIV and Alzheimer's treatment, safety and inadequacy concerns in the gene

editing should be prioritized.

The bottom line in the treatment of HIV and Alzheimer's using the CRISPR-Cas9

remains a promising strategy that should be researched and developed further. However, research

and development still plays an integral part in overcoming the associated obstacles before the
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technique could be widely used. Overcoming delivery barriers, controlling off-target effects,

addressing ethical concerns, ensuring the long-term viability of modified cells, and controlling

the complexity of HIV infection are important areas that still need research and improvement.

The researchers are also considering the use of small molecule medication, lentiviral vectors

application on gene therapies and antibody therapies besides CRISPR-Cas9 on the treatment of

HIV and Alzheimer's disease. Lastly, the treatment of HIV and Alzheimer's highly depends on

the successful research and development of CRISPR-Cas9 to be able to overcome challenges and

obstacles. However, it is important to consider the alternatives and have autonomy of choices

when it comes to fight against HIV and Alzheimer's.

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