ISSN 0006
2979, Biochemistry (Moscow), 2013, Vol. 78, No. 1, pp. 1
7. Pleiades Publishing, Ltd., 2013.

Original Russian Text O. N. Gusachenko, M. A. Zenkova, V. V. Vlassov, 2013, published in Biokhimiya, 2013, Vol. 78, No. 1, pp. 5
13.

REVIEW

Nucleic Acids in Exosomes: Disease Markers

and Intercellular Communication Molecules
O. N. Gusachenko*, M. A. Zenkova, and V. V. Vlassov
Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, pr. Lavrentieva 8,
630090 Novosibirsk, Russia; fax: (383) 363
5153; E
mail: niboch@niboch.nsc.ru; oligonucleotide@gmail.com
Received July 2, 2012
AbstractThe term exosomes is currently used to describe specific vesicular structures of endosomal origin produced by
the majority of eukaryotic cells. These natural vesicles have been under study for more than two decades. Nevertheless, a
real splash of scientific interest in studies on exosomes took place only during recent years, when the concept of the role and
functions of exosomes in multicellular organisms was essentially reconsidered. The major role in this was played by the dis

covery of exosomal mRNA and miRNA in 2007, which stimulated the idea of regulatory and communicative role of exo

somes in the organism and also encouraged considering exosomes and other vesicles as potential biomarkers. The present
review summarizes the up to date knowledge on the composition and probable physiological functions of nucleic acids
released by different cells as components of exosomes. We also touch upon the problem of using these data in clinical diag

nosis.
DOI: 10.1134/S000629791301001X
Key words: exosomes, microvesicles, extracellular nucleic acids, biomarkers, intercellular communication, miRNA, mRNA

Exosomes are small vesicular bodies of 50
90
nm

size [1] (or 30
100 nm according to another reference [2])
formed from late endosomes. The formation of the exo

some starts from an inner invagination of the endosomal
membrane that results in a luminal vesicle, whereas the
endosome becomes a multivesicular body. The fusion of
late endosomes with the cytoplasmic membrane is associ

ated with release of luminal vesicles into the extracellular
space. Thus, exosomes are luminal vesicles deserted from
the cell [3]. The exosomal cavity has cytoplasmic origin,
whereas their membrane is a result of invagination of the
endosomal membrane. Theoretically, any cell containing
multivesicles can secret exosomes. In fact, according to
experimental data, exosomes can be produced in vitro by
the majority of differentiated cells: the exosome produc

tion by B
and T
cells, granulocytes, dendrite cells,
platelets, neurons, and epithelial cells is described rather
in detail. Exosomes were also found in vivo, in particular,
they are present in many physiological liquids, including
blood serum, bronchoalveolar lavage, urine, and breast
milk [4]. The work published in 1983 on the fate of trans

ferrin receptors during maturation of sheeps reticulo

cytes is usually considered to be the first scientific

description of exosomes [5]. The authors found that dur

ing the maturation of reticulocytes, exosomes could par

ticipate in restructuring of membranes by carrying the
transferrin receptors out [6]. Initially researchers consid

ered exosomes as either formations specific for a strictly
definite type of cells or as byproducts of the cells vital
activity free of any significant function. The discovery
that antigen
presenting cells can secret exosomes pos

sessing their own immunostimulating activity changed
the attitude of scientific community to these vesicles [7].
Valadi et al. described in 2007 a large pool of RNA mole

cules found in exosomes of mastocytes [8]. It was this
publication that moved the studies in the field on a fun

damentally new level.
Each new article now adds surprising examples to the
set of supposed functions of exosomes. Exosomes are
involved in elimination of unwanted proteins from the
cell membrane and contribute to initiation and develop

ment of the immune response. Leukocytes use the release
of a huge amount of exosomes during coagulation.
Exosomes can be involved in development of some patho

logical processes, such as growth and spreading of malig

nant tumors, transfer of infectious agents whole virus
particles or viral RNAs, and also of prions. Exosomes
seem to play a special role in the intercellular communi

cation of multicellular organisms as they carry on their

* To whom correspondence should be addressed.

GUSACHENKO et al.

surface various protein and lipid ligands and also deliver

new proteins and nucleic acids into recipient cells [9].
Exosomes have been found to contain different RNAs,
including messenger RNA (mRNA), microRNA
(miRNA), and long non
coding RNA. The main func

tion of mRNA in the organism is the transmission of
information about the amino acid sequence of a protein
to the synthetic machinery of the cell. The transfer of
intact mRNAs as parts of exosomes can serve for
exchanging phenotypic features between cells, because
recipient cells get mRNAs of proteins not expressed in
them initially. The miRNAs are short non
coding RNAs
of 21
25 nucleotides length. They are involved in regula

tion of gene expression on posttranscriptional level by
triggering mechanisms of translation blocking or destruc

tion of mRNAs containing complementary or partially
complementary sequences. In some cases, miRNAs can
regulate gene expression also on the level of transcription,
influencing the modification of histones and methylation
of promoter regions of DNA [10]. Supposedly, the
miRNA targets are present in approximately 60% of all
mammalian genes. In any cell of the body, there are large
amounts of miRNAs, and each miRNA is able to influ

ence hundreds of mRNA targets. The miRNAs are
known to be involved in regulation of various processes
including the cell differentiation, division, apoptosis,
endocrine system functioning, hemopoiesis, and mor

phogenesis of different organs [10]; the role of miRNAs
in development of various pathological processes, such as
cancer, viral diseases, hereditary diseases, etc. is also
under active investigation.
The work of Ratajczak et al. on cultivation of embry

onic stem cells [11] was one of the first demonstrations of
horizontal mRNA transfer between cells [11]. The
authors found that vesicles produced by these cells could
transfer specific mRNAs into precursors of hematopoiet

ic cells, which resulted in changes of cells phenotype.
Later the presence of extracellular miRNAs in various
biological fluids of the body was shown [12]. Based on the
available experimental data, it was supposed that nucleic
acids transferred in the exosomal cavity should have an
epigenetic influence on the organisms cells. This
hypothesis not only changed ideas about intercellular
communication mechanisms, but it also stimulated stud

ies on possible use of exosomal RNAs as biomarkers.

METHODS FOR ISOLATION OF EXOSOMES

In the literature the terms exosome and
microvesicle often overlap, and sometimes they are
even used as synonyms. In the first place, this is due to
preferential use by authors of the more generalized term
microvesicles that also includes larger membrane for

mations (more than 1 m), instead of a strict determina

tion of the type of vesicles under study. Differences in the

initial biological materials and also different approaches

and goals of studies have resulted in a rather confused ter

minology, and within its limits vesicular particles pro

duced by cells can be designated in various ways, e.g.
microparticles, microvesicles, ectosomes or shedding
microvesicle, nanovesicles (nanoparticles), exosomes,
exosome
like particles, apoptotic blebs, promini

nosomes, prostatosomes, dexosomes (dex), texosomes
(tex), epididimosomes, argosomes, archaeosomes, or
oncosomes [13]. Most strictly, the term exosome desig

nates the membrane particles of endocytotic origin with
size less than 100 nm that are produced as a result of
fusion of multivesicular bodies with the plasma mem

brane.
The classical protocol for isolation of exosomes
includes a number of successive centrifugations that
allows to get rid of cells and larger particles, and then exo

somes are precipitated by ultracentrifugation at 100,000g
[14]. In this case, the desired particles are subtracted from
a sample by their size. Thus, this approach is not specific
for vesicles of strictly endosomal origin. A very simple
additional procedure of filtration through small
porous
filters (0.1
0.2 m) can free the final precipitate from
larger particles (microvesicles) [15], but it cannot sepa

rate exosomes from other particles with similar size (pro

tein aggregations). Exosomes can be additionally purified
by isolation in a sucrose gradient or on a sucrose cushion.
Exosomes are known to float at the density range from
1.13 g/ml (products of B
cells) to 1.19 g/ml (products of
intestine cells) [14]. This purification results in a suffi

ciently effective separation of exosomes from unassociat

ed protein complexes and fragments of nucleosomes
released by apoptotic bodies.
The use of successive centrifugation for isolation of a
relatively small quantity of exosomes is a rather inconven

ient and laborious procedure. Lamparski et al. worked out
a protocol for isolation of clinical grade exosomes in
which the precipitation stage was replaced by an ultrafil

tration procedure with subsequent purification of the
preparation on a sucrose cushion. The authors proposed
this method as more rapid, productive, and reproducible
than the classic one [16]. In work [17], Cheruvanky et al.
used a commercial nanomembrane concentrator to pre

pare exosomes suitable for clinical analysis, and this also
allowed them to avoid the ultracentrifugation. In addition
to filtration, exosomes can be isolated from specimens
under analysis by precipitation in the presence of differ

ent polymers (as done for isolation of viral particles) with
a subsequent filtration or centrifugation. Supposedly, this
principle is used in ExoQuick product (System
Biosciences, USA; http://www.systembio.com) designed
for exosomes isolation from urine or blood serum.
However, despite the declared efficiency, according to the
users references this preparation can precipitate not only
exosomes but also other particles, and this creates diffi

culties in its use.
BIOCHEMISTRY (Moscow) Vol. 78 No. 1 2013

NUCLEIC ACIDS IN EXOSOMES

Using antibodies against exosomal markers or pro

teins specific for definite cell types seems to be a promis

ing approach for selective isolation of exosomes of known
nature. Depending on the purpose and methodology of
the study, antibodies can be immobilized on magnetic
beads, chromatographic matrices, plates, or other work

ing surfaces. This approach for isolation results in prepa

ration of exosomes with a known set of antigens, but the
subsequent washing of exosomes from the antibodies can
be rather inefficient, and this limits their further applica

tion. Flow cytometry of exosomes on beads is a conven

ient method for detecting the known protein markers in
the preparation and serves as additional method of quali

ty control. Currently other affinity approaches for isola

tion of exosomes including filtration systems or capture
on affinity columns are under development [18]. Using
lectins capable of binding sugar residues exposed on the
surface of vesicles seems to be a new interesting approach
in this field (http://www.aethlonmedical.com).
Vesicles can be characterized as exosomes based on a
combination of morphological, biochemical, and physi

cal properties. Because of their small size, exosomes can
be observed only by electron microscopy [19]. The quali

ty of an exosome preparation can be also assessed by sep

aration in polyacrylamide gel with subsequent specific
and nonspecific staining of proteins. Many proteins are
accumulated in exosomes selectively (e.g. flotillin, Alix,
CD63) and can be used as markers of the exosomal frac

tion. Exosomes are also characterized by saturation of
their membrane with cholesterol, sphingolipids, and
glycerolipids that makes them similar to lipid rafts.
Exosomes of some cells contain not only protein and
membrane components, but also different types of RNA,
which can be easily isolated and identified by standard
analytical methods of molecular biology. It is rather prob

able, that the presence of nucleic acids in exosomes can
also be used for detection of the exosomal fraction in a
preparation and for identification of exosomes with a def

inite histological origin.

RNA AS A COMPONENT OF EXOSOMES

In the work published by Valadi et al. in 2007 about
1300 different mRNAs and 121 miRNAs were mentioned
(according to the database http://mirbase.org there are
about 1500 different miRNA known in human to the
date). Later, RNAs were found in exosomes of various
cells (mast cells, tracheobronchial cells, dendrite cells, B

and T
lymphocytes, cells of intestine, lung, and stomach
cancers, and of pancreatic adenocarcinoma [20
23]) and
also in exosomes isolated from different bodily fluids
(blood, saliva, breast milk, pleural fluid, ascites [24, 25]).
By the initiative of Simpson and Mathivanan, the elec

tronic database ExoCarta (http://exocarta.org) was creat

ed, which includes information about proteins, lipids,
BIOCHEMISTRY (Moscow) Vol. 78 No. 1 2013

mRNAs, and miRNAs of exosomes with different origin.

According to the data of ExoCarta, in the majority of the
published works new RNAs were identified using
microarray methods. Now several thousands of different
RNAs are described as presumable components of mam

malian exosomes (more than 1600 mRNAs and more
than 700 miRNAs according to ExoCarta).
On appearance of the first data about exosomal
RNAs, their presence in the exosome preparations was
initially supposed to be due to contamination of the spec

imens by molecules of nucleic acids released from dying
cells. On the other hand, different RNAs could be
enclosed in an exosome during its formation due to occa

sional capture of molecules occurring in the cytoplasm of
the cells. According to data published by Valadi et al. [8]
and also by several other groups, concentration ratios of
many mRNAs found in the exosome preparations signif

icantly differ from the ratios in the cells cytoplasm. The
results of Ohshima et al. revealed a selectivity of miRNAs
package in exosomes of metastasizing intestinal cancer,
whereas the initial poorly metastasizing cell line did not
display a similar selectivity. The authors concluded that in
this case the secretion of miRNAs by the exosomal path

way could play a role in oncogenesis [21]. The existence
of a specific mechanism of RNA sorting for directed
secretion was suggested. An attempt to perform comput

er
aided analysis of exosomal RNAs was performed by
Batagov et al. [26]. They reported the presence of some
specific repeats in sequences of excreted RNAs that could
act as cis
elements under selective conditions. In the case
of experimental confirmation of the data, this study can
at least partially dispel the mystery of the mechanism of
RNA excretion via exosomal pathway. In any case, no
data was actually found to the date in favor of existence of
specific exosomal RNA sequences. On the contrary, it
was shown for some tumors that exosomes can contain
sets of miRNAs with the composition similar to that in
the initial cancer cells [27, 28]. Although some data indi

cate differences in the pools of intracellular and excreted
RNAs, it is still unclear whether such sorting is selective.
But how the sorting of RNAs into exosome is real

ized? It was found that multivesicular bodies and effector
complexes of miRNAs are functionally related [29, 30].
Such co
localization seems to indicate that multivesicu

lar bodies are places for assembly of miRNAprotein
complexes, and during this process miRNAs also can be
selected for secretion in exosomes. Kosaka et al. [31]
showed that the release of exosomal miRNAs can be con

trolled by sphingomyelinase 2 and realized via a
ceramide
dependent pathway. This corresponds with the
observations of Mittelbrunn et al., showing that inhibi

tion of sphingomyelinase expression leads to disturbance
in the miRNA transfer into antigen
presenting cells and
that ceramide is involved in triggering miRNA secretion
by the cells [23]. Montecalvo et al. [32] studied the mech

anism of exosomal miRNA transfer between dendritic

GUSACHENKO et al.

cells. Exosomes isolated from the culture of dendritic

cells contained 200 miRNAs, and five of them were pres

ent only in exosomes of immature dendritic cells, where

as 58 miRNAs were found only in exosomes of mature
dendritic cells. Thus, the authors showed that dendritic
cells can produce exosomes with different RNA composi

tion depending on the stage of their developmental. The
set of miRNA present in exosomes was also different from
that in the initial cells. The functional activity of two
miRNAs transferred by exosomes was confirmed in this
study using the transfection of treated cells with luciferase
conjugated target.
The fact that nucleic acids can be captured into exo

somes and that their presence inside the vesicles does not
seem to be accidental suggests the supposition on the
probable role of exosomes in the horizontal transfer of
genetic information between cells of a multicellular
organism. In particular, exosomes can be unique trans

ducers of signaling information between distant cells and
tissues. Thus, mRNA inside exosomes of mastocytes was
shown to be translationally active, i.e. on entrance into
recipient cells this RNA could participate in the synthesis
of new proteins [8]. The authors also found that such
transfer of information can occur in vitro between the
cells of two different organisms mouse and human:
upon incubation of a culture of human mastocytes in the
presence of mouse exosomes, mouse proteins were
detected in the human cells, and three of these proteins
were absent in the exosomes themselves, but their
mRNAs were found in the vesicles. The mechanism of
interaction between cells and exosomes is still under
investigation. The difficulty of these studies is associated
in particular with the extremely small size of the exosome,
which prevents observations with a fluorescence micro

scope. The transfer of material through exosomes
between cells can also vary depending on the type and
state of the cells. Thus, if an interaction of an exosome
with a cell occurs with involvement of surface receptors,
we may speak about a specific directed delivery of the
exosomal material into strictly definite types of cells. The
possible existence of such mechanism was also confirmed
by data of Valadi et al. [8]. They found that exosomal
RNA could be transferred between mastocytes, but it did
not penetrate into CD4 lymphocytes.
The transport of nucleic acids inside the exosomes
can play an important role in functioning of a multicellu

lar organism; moreover, the supposed specificity of this
process can, on necessity, contribute to the directed
transfection of an exogenous material. Alvares
Erviti et
al. [33] used exosomes of mice dendritic cells as a trans

fection vector for short interfering RNAs. It was supposed
that the use of exosomes produced by animals own cells
will lead to the decreased immunogenicity of this vector,
whereas inclusion into exosomes of a hybrid protein con

sisting of Lamp2b (a membrane protein of the exosomal
fraction) and a neuron
specific peptide RVG should

result in a directed delivery of RNA into the cells of nerv

ous tissue. The intravenous injection of exosomes electro

porated with delivered RNAs resulted in a specific deliv

ery of biologically active RNAs into neurons, microglia
cells, and brain oligodendrocytes, but not into other tis

sues (liver, spleen, kidneys, etc.). The exosome
mediated
transfection allowed the authors to obtain significant
decrease of the levels of the target mRNA and of the pro

tein encoded by it (by 60 and 62%, respectively). In this
case the authors showed not only the possibility of using
short RNAs inside exosomes as a transfecting system
capable of inhibiting the target gene expression in brain
tissues (i.e. of penetrating across the bloodbrain barri

er), but the principle of in vivo functioning of exosomes
was demonstrated, which is of no less importance.
What possible biological role the exosomal RNAs, or
shuttle RNAs (shRNAs) as they are often referred to,
might play? According to analysis performed with the
Ingenuity system software (http://www.ingenuity.com),
mRNA
transcripts from exosomes of mastocytes are
functionally related to the processes of cytogenesis, pro

tein synthesis, and posttranscriptional modification of
RNA [8]. Analysis of miRNAs found in exosomes of these
cells is more complicated, first of all because of multiplic

ity of miRNA functions. The noticeable difference
between the miRNA pool in the exosomes of mastocytes
and in the cytoplasm probably indicates a certain direct

ed regulatory function of these RNAs inside the vesicles.
The identified miRNAs include let
7, lin
4, miR
1, miR

15, miR
16, miR
17, miR
18, miR
181, miR
375, etc.
These miRNAs are involved in the regulation of cell
development, angiogenesis, hematopoiesis, exocytosis,
and oncogenesis. The released miRNAs are now consid

ered to represent a new class of paracrine regulation mol

ecules. A possible example of this function might be an
involvement of exosomal miRNAs in the regulation of
cardiomyocyte growth. It was shown that the miRNAs
from fibroblasts can penetrate into cardiomyocytes and
induce the development of hypertrophy [34].
Regulation of the immune system can be an impor

tant function of released miRNAs. Both T
and B
lym

phocytes and also dendritic cells of the immune system
are known to secrete miRNAs as components of exo

somes [23]. It was also shown that the interaction of T

lymphocytes with antigen
presenting cells could be asso

ciated with antigen
induced directed transfer of
miRNAs. Exosomal miRNAs are transferred into anti

gen
presenting cells during formation of the immune
synapse with T
lymphocyte, and they can function in the
recipient cell. The involvement of exosomes in the trans

fer of biologically active RNAs from mother to child dur

ing pregnancy and breast
feeding might represent a
unique example of immunological functions of exosomes.
Specific placental miRNAs were detected in exosomes of
the villiferous trophoblast [35]. The exchange of exoso

mal contents between the fetus and mothers tissues is
BIOCHEMISTRY (Moscow) Vol. 78 No. 1 2013

NUCLEIC ACIDS IN EXOSOMES

supposed to provide adaptation of the two organisms to
one another during the progression of pregnancy. Breast
milk also contains a significant amount of immuno
rele

vant miRNAs, and at least some of them are inside the
exosomes [36]. These miRNAs might play an important
role in development of the childs immune system.
In addition to involvement in normal functioning of
the body, exosomal RNAs can also play a crucial role in
development of health disorders. Progression of oncolog

ical diseases directly depends on the efficiency of com

munication between the malignant and healthy cells.
Primary glioblastoma cells were shown to secrete micro

vesicles and exosomes containing mRNAs and miRNAs
[37]. It was found that a culture of multivesicular brain
cells is capable of taking up these vesicles, which resulted
in translation of the transported mRNAs. Thus, using the
exosomal pathway, tumor cells could modify the transla

tion profiles of surrounding healthy cells. Oshima et al.
[21] suggested that exosomal RNAs could have an onco

genetic function. They found that miRNAs of the let
7
family were intensively excreted inside exosomes of the
AZ
P7a cell line from metastasizing intestinal cancer.
Because miRNAs of the let
7 family usually mediate the
suppression of tumor growth, the authors supposed that
secretion of these RNAs inside the exosomes allowed the
tumor to decrease the anti
oncogenic activity of the cells
and increase the probability of the tumor metastasis.
Exosomes are also known to promote distribution
and maintain the populations of some pathogens includ

ing certain viruses and prions [38]. Type
4 herpes virus, or
EpsteinBarr virus, is considered to be one of the most
widespread human viruses. On the grounds of exosomal
RNA profiles analysis for infected cells, Pegtel et al.
detected in the vesicles virus
encoded miRNAs [39]. This
work was performed on a co
incubation model using
infected B
lymphoid cells with a culture of dendritic
cells. The results revealed an ability of the viral miRNAs
to be transferred into non
infected cells and display there
their biological activity. While the viral DNA was found
only in the B
lymphocyte population, miRNAs of the
EpsteinBarr virus were detected not only in the infected
B
lymphocytes, but also in other (non
infected) cells of
patients with an increased level of the virus in the body.
These data on transduction of viral miRNA into non

infected cells confirm the hypothesis about the evolution
of herpes viruses in the direction of exploitation of the
hosts cells interference machinery.

EXOSOMAL NUCLEIC ACIDS AS BIOMARKERS

The cells in the organism release exosomes into the
extracellular space normally and also on development of
diseases. These vesicles contain macromolecules of pro

teins, lipids, and nucleic acids that can be used as bio

markers of the producing cell state. The presence of a sig

BIOCHEMISTRY (Moscow) Vol. 78 No. 1 2013

nificant amount of exosomes in physiological fluids of the

body (blood plasma, urine, breast milk, sperm, saliva,
etc.) provides the opportunity to use noninvasive or lowly
invasive approaches for sampling; therefore, these parti

cles are a desirable object for development of approaches
for routine clinical analyses. Exosomal nucleic acids pos

sess many features of good biomarkers. The excreted
nucleic acids are highly stable in biological fluids (this
observation is often used to indirectly confirm the release
of nucleic acids as parts of special protective complexes).
The levels of RNAs in preparations can be easily deter

mined by standard methods of molecular biology based on
quantitative PCR, which possess high sensitivity and effi

ciency. Strictly speaking, RNAs can be found in physio

logical fluids not only in exosomes, but also in other vesi

cles, as well as in complexes with proteins. Comparison of
mRNA pools in exosomes and microvesicles of similar
origin (or in combined specimens of exosomes and
microvesicles) might be of particular interest [29]. At pres

ent, there is no uniform opinion on the main form of RNA
transfer in intercellular fluids: for example, the results of
Turchinovich et al. [40] show that the major part of extra

cellular RNA is vesicles
free. In our opinion, it is reason

able to suggest that the release of RNA can exploit differ

ent mechanisms and depends on the secretion conditions
(developmental state of the cell, macro
and microenvi

ronment, presence of pathology, general state of the
organism) and on the type of RNA targets (i.e. what form
of the transported RNA can be taken up by the recipient
cell). However, the lack of clear knowledge on this subject
does not prevent many medico
biological companies from
the initiation of new projects, which include the use of
exosomal nucleic acids in diagnostics. Exosomes have
been shown to contain tissue
or tumor
specific sets of
miRNAs that seems to be promising for diagnosis. Before
the discovery of exosomal RNAs, the analysis of tumor
miRNA profile usually required biopsy an invasive pro

cedure, which might be strongly undesirable or even
impossible in certain types of cancer. Application of RNA
profiles analysis can be preferable in diagnostics of the
tumors which do not possess any known and reliable
molecular markers, e.g. in ovarian cancer [37]. Skog et al.
were first to demonstrate that secreted miRNAs can be
used for diagnosis of glioblastoma and determining the
stage of desease [37]. It was found that the level of exo

somes in blood serum of patients with ovarian cancer cor

related with the disease stage; moreover, the total number
of exosomes in the patients blood was higher than in the
control group [27]. The authors also reported that miRNA
profiles in specimens of the exosomes were close to the
miRNA set of the tumors. Similar results were obtained in
a study on RNA composition of exosomes of healthy
donors and patients with lung cancer [28]. The knowledge
on the presence of viral RNAs in the exosomes (e.g. exo

somes of type
4 herpes virus) can also be applied for the
infection detection in the organism.

GUSACHENKO et al.

Sjogrens syndrome is an autoimmune systemic dis

ease manifesting itself in particular by chronic influence
on the salivary and lachrymal glands. Michael et al. isolat

ed exosomes from the saliva of patients with this syndrome
and showed that these vesicles contain significant amounts
of miRNA. On the grounds of obtained data, the authors
suggested the possibility of new diagnostic method elabo

ration for the asymptomatic patients [41]. In this case, the
noninvasive and rapid analysis of the patients saliva can
be a good alternative for the routinely used biopsy.
Urine is another easily accessible fluid naturally
released from the body. Tests using exosomes in urine are
now actively developed for diagnosis of urogenital dis

eases, in particular, of prostate cancer [42]. Two specific
mRNAs (PCA3 and a product of specific chromosome
aberration TMPRSS2:ERG), which are hyperexpressed
on prostate carcinoma, were found in exosomes of the
patients urine.
Other prospects of using exosomal RNAs as bio

markers include diagnosis of probable traumas of the
brain by the presence of specific vesicular miRNAs in the
cerebrospinal fluid, the early detection of vascular dis

eases, and also diagnosis in the case of asymptomatic dia

betes based on specific profiles of blood miRNAs. Finally,
the diagnostic potential of exosomal biomarkers
embraces not only pathological processes. Thus, compar

ison of miRNA levels in blood plasma of pregnant and
non
pregnant women revealed the correlation with the
placenta
associated miRNAs and the presence of preg

nancy and also with its stage [43].
The concept of intercellular communication via
directed and selective transfer of genetic material by exo

somes is very prospective for modern biology and medi

cine. It should be emphasized that the overwhelming
majority of experimental data describing various process

es involving exosomes in vitro and in vivo has been
obtained with microparticles isolated from cell cultures.
The correlation between the conclusions based on these
data and the real principles of exosome functioning under
conditions of a living body still relates to one of the most
significant issues in this field. Nevertheless, this does not
diminish the value of the obtained results, as they direct

ly indicate the practical potential of exosomes applica

tion. Although current knowledge fails to give a complete
idea about the origin and nature of RNA
containing vesi

cles, many pharmacological companies are intensively
working to create test systems based on exosomes. In
some laboratories, the contents of exosomes (and also of
other extracellular vesicles) are comparatively analyzed in
specimens obtained from healthy donors and patients
with different diseases. There are many prerequisites
favoring the application of exosomal RNAs as biomark

ers. Among the problems, which are to be solved by the
test system developers, lies the necessity to elaborate an
effective and economical isolation method of exosomes

(or their components, e.g. RNA) from small volumes of

biological fluids. Another issue is standardization of
methods for RNA analysis in specimens, including the
search for stable control biomarkers with low variability in
the population and high reproducibility in standard ana

lytical procedures. Considering the amount and intensity
of modern studies in this field, it will be possible to verify
the efficiency of currently developing approaches within
1
2 years. In which areas of diagnostics the exosomes
based methods will be actually applicable practice and
further studies will show.
The work was financially supported by RFFI (grant
No. 11
04
01429
a), program Molecular and Cellular
Biology of the Presidium of the Russian Academy of
Sciences, and Siberian Branch of the Russian Academy
of Sciences (project No. 84).

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