ROLE OF EXOSOMES IN ALZHEIMER’S DISEASE

1. INTRODUCTION

Alzheimer's disease (AD) is a chronic, progressive and irreversible neurodegenerative

disorder that causes decline in cognitive function as well as behavioral and social
performance deterioration[1]. According to the latest estimations, Alzheimer's disease is the
most common cause of dementia, accounting for 60% and 80% of cases each year[2]. It is a
potentially fatal situation for the elder people, as well as a social and financial burden on
families and the healthcare system [3]. Alzheimer's disease is an aging disorder characterized
pathologically by cerebral amyloid-beta (Aβ) plaques and intracellular neurofibrillary tangles
(NFTs) [4]. It has also been proposed that Alzheimer's disease is linked to the aggregation of
tau proteins. Changes in tau protein phosphorylation impair its ability to stabilise tubulins,
resulting in microtubule disorganisation[5]. Neurofibrillary tangles are formed as a result of
tau protein aggregation. Neurofibrillary tangles consist of hyperphosphorylated tau proteins
found in areas of the brain responsible for learning, memory, and emotion. Although these
features are considered characteristic pathologic hallmarks of the disease[6]. Alzheimer's
disease (AD), neurobiological brain disorder that gradually kills brain cells, causes memory
and cognitive capacity deficits, and ultimately speed up the loss of ability to perform even the
most basic tasks [7]. Alois Alzheimer described the disease for the first time in 1906, he
described a 50-year-old woman suffering from an unusual and progressive illness
characterized by aggression, confusion, and memory loss[8]. She died 5 years later, and at
autopsy, Alzheimer discovered shrinkage of the brain as well as abnormal deposits, which he
named plaques and neurofibrillary tangles. Since then, there have been many technological
advances that have aided in unlocking the secrets of this devastating disease [9]. The clinical
manifestations of Alzheimer's disease, such as memory loss and altered cognitive functions,
are caused by synaptic and neuronal loss, as well as a prominent inflammatory state in the
entorhinal cortex, hippocampus, and neocortex[10]. The disease is divided into 2 subtypes
based on the age of onset: early-onset AD (EOAD) and late-onset AD (LOAD). EOAD
affects between 1% to 6% of the population, with ages ranging from 30 to 60-65 years[11].
However, the most frequent form of AD, LOAD, is described as AD that begins after the age
of 60 or 65 years[9]. People with a positive family history of AD are more likely to develop
both EOAD and LOAD[12]. Genetically, AD is usually divided into familial cases with
Mendelian inheritance (Familial AD, FAD) and sporadic cases with no familial
aggregation[13].Mutations in three major genes cause the familial form of Alzheimer's
disease: amyloid precursor protein (APP), presenilin 1 (PSEN1), and presenilin 2 (PSEN2)
[14] . Mutations in these genes increase the production of Aβ from APP (Amyloid precursor
protein) via sequential cleavage by β- and γ-secretases, causing neuronal death and the
accumulation of amyloid plaques in the brain [15]. The main components of amyloid plaques
are two abnormally folded APP metabolism products: Aβ40 and Aβ42, which have a higher
rate of oligomerization and insolubility[16]. The researchers discovered that approximately
60% of people with Alzheimer's disease carry the gene apolipoprotein E4 or ApoE4[17].
Apolipoprotein E (APOE) is the primary apolipoprotein found in the brain and is essential for
lipid transport and homeostasis. Another gene, SORL1, appears to be frequently involved as
well[18]. However, many people who carry these genes will never develop the disease, while
others who do not carry these genes may develop the disease[19]. Allelic abnormalities on
the APOE-gene on chromosome 19 are best known for predicting the onset and increasing the
severity of both inherited and sporadic Alzheimer's disease [20]. Different isoforms of the
ApoE gene in humans have been linked to the occurrence of sporadic Alzheimer's disease.
Apolipoprotein E (APO-E) binds to the β-amyloid peptide and is found in senile neuritic
plaques in Alzheimer's disease [21]. These isoforms differ only in two amino acids and are
known as ApoE 2 (Cys112, Cys158), ApoE 3 (Cys112, Arg158), and ApoE4 (Arg112,
Arg158), which differ by the presence of cysteine or arginine at specific residues [22]. ApoE
4 has been linked to both sporadic and familial late-onset Alzheimer's disease, implying a
causal role . Moreover, the APOE4 allele is a factor that modifies the age of onset in EOAD
caused by APP and PSEN2 mutations [23]. Although APOE is the well-studied genetic
contributor to Alzheimer's disease, the exact molecular mechanisms underlying its effects on
disease onset remain unknown[24]. A variety of lifestyle factors, including diet, smoking, and
physical activity, may interact with the APOE genotype to influence cognitive
impairment[25]. In contrast, the vast majority of Alzheimer's cases (95% to 90%) were
discovered to be late-onset and sporadic. Susceptibility genes may play a role in the
development of sporadic Alzheimer's disease (SAD) [26]. Other pathological hallmarks of
AD include neuronal loss in specific brain areas, neuroinflammation, oxidative stress and
metal dyshomeostasis[27] .

2. EXOSOMES BIOGENESIS AND PHYSIOLOGICAL FUNCTION

The term exosome was first used in the early 1980s to describe vesicles that were released by
various types of cultured cells and ranged in size from 40 to 1000 nm[28]. Over the next ten
years, this term came to be used to describe endosomal vesicles ranging in size from 30 to
100 nm[29]. Exosomes are extracellular vesicles (EV) that are secreted by all brain cells,
including neurons, microglia, and astrocytes, and play an important role in cell-cell
communication[30]. Apart from exosomes, the two other major subtypes of EVs are
microvesicles and apoptotic bodies. Exosomes are the smallest EVs with an endocytic origin,
whereas microvesicles (50-1000 nm) are formed by outward budding from the plasma
membrane and apoptotic bodies (500- 4000 nm) are derived from apoptotic cells[31].
Exosomes are formed when multivesicular bodies (MVBs) fuse with membranes. Indeed,
early endosomes mature into late endosomes, where they form intraluminal vesicles (ILVs)
within MVBs [32]. When ILVs form, they secrete cytoplasmic molecules such as proteins,
messengerRNA (mRNA), and small non-coding microRNAs (miRNAs) within them and
accumulate inside the late endosome, resulting in the formation of MVB [33]. MVBs can fuse
with lysosomes (resulting in the degradation of their contents) (dMVBs) or with the plasma
membrane, releasing their contents, the exosomes, into the extracellular space (sMVBs)[34].
In comparison to dMVBs, which are high in bismonoacylglycerophoshate (BMP, LBPA),
sMVBs are higher in ceramides. The formation of ILVs is a key step in the biogenesis of
exosomes[35]. The formation of ILVs is primarily regulated by the Endosomal Sorting
Complex Required for Transport (ESCRT) complex of multi-molecular machinery. However,
studies have shown that depletion of ESCRT subunits does not completely impair the
composition of MVBs, implying that other mechanisms may be involved in the formation of
ILVs[36]. It was suggested that proper levels of lipids and tetraspanin-enriched micro-
domains are required for the formation of MVBs. Membrane depolarization also influences
exosome secretion. By the end of the twentieth century, it was discovered that B-lymphocytes
and dendritic cells secreted comparable vesicles, and most cell types, including those in the
central nervous system (CNS), are now known to release exosomes[37]. Exosomes are
secreted by reticulocytes, mesenchymal cells, neurons, fibroblasts, epithelial cells, endothelial
cells, platelets, allophycocyanins (APCs), tumour cells, and other cells, and are found in
bronchioalveolar lavage, synovial fluid, urine, bile, breast milk, serum, and other body fluids
because they display specific cell-derived components on their surfaces exosomes from
various cell types contain the same set of molecules, including enzymes, nucleic acids,
cytokines, and bioactive compounds [38]. Exosomes are secreted in both physiological and
pathophysiological conditions, and their actions on neighbouring and distant cells lead to the
modulations of cellular behaviours [39]. Exosomes not only play a housekeeping role in
maintaining normal cellular functions and viability, but they also protect various functions of
multicellular organisms[40].

3.EXOSOMES IN ALZHEIMER’S DISEASE

AD arises as a result of the extracellular deposition of amyloid-β (Aβ; encoded by AβPP)

fibrils and of abnormally phosphorylated tau protein (encoded by MAPT) in neurons Aβ
propagation via exosomes to extracellular milieu was reported by (Rajendran et al.
2006). who observed cleavage of APP followed by secretion of β-amyloid in exosomes.
Subsequently, secretions of the C-terminal part of APP was observed in vitro  and in vivo .
Though, exosomes have been reported to induce the formation of neurotoxic oligomers of Aβ
, they have also been reported to have a neuroprotective role of clearing toxic oligomeric
species in exosomal lumen. Impaired Aβ clearance in AD patients and the neuronal and
microglial exosome disposal of Aβ hints at the possibility of a dual clearance mode for Aβ in
AD. Exosomal proteins such as Alix and Flotillin-1 were also found to be accumulated in the
plaques of AD patient brains. Neurofibrillary tangles representing hyper-phosphorylated
misfolded tau have been observed in the CSF of AD patients . APP, β- secretase, γ- secretase
has been detected in exosomes APP undergoes enzymatic cleavage by β and γ secretases to
generate Aβ peptide, which is released into the extracellular space. Because its clearance
from the brain slows with aging, Aβ peptide gradualy accumulates and ultimately results in
the characteristic amyloid plaque formation observed in AD patients. Exosomal secretion of
tau is considered critical for the spread of tauopathy to different areas of the AD brain.
Contrary to the dependence of tau propagation on the exosomal secretory pathway , recent
studies on the role of exosomes in proteinopathies revealed regulation of Tau secretion by
neuronal activities.

4.miRNA-MEDIATOR OF AD

5.EXOSOMES AS BIOMARKERS IN AD
6.EXOSOMES-OXIDATIVE STRESS

7.EXOSOMES-ANGIOGENESIS

8.EXOSOMES-INFLAMMATION

9.CONCLUSION AND FUTURE PROSPECTIVE

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