Abstract
The
integration
of
robust
single‐pot,
solid‐phase‐enhanced
sample
preparation
with
powerful
liquid
chromatography‐tandem
mass
spectrometry
(LC‐MS/MS)
is
routinely
used
to
define
the
extracellular
vesicle
(EV)
proteome
landscape
and
underlying
biology.
However,
EV
studies
are
often
limited
by
availability,
requiring
upscaling
cell
cultures
or
larger
volumes
biofluids
generate
sufficient
materials.
Here,
we
have
refined
data
independent
acquisition
(DIA)‐based
MS
analysis
optimizing
both
protein
enzymatic
digestion
chromatography
gradient
length
(ranging
from
15
44
min).
Our
short
min
can
reproducibly
quantify
1168
(from
as
little
500
pg
peptides)
3882
proteins
groups
50
ng
peptides),
including
quantification
22
core
marker
proteins.
Compared
data‐dependent
acquisition,
DIA
achieved
significantly
greater
coverage
low
abundant
species.
Moreover,
optimal
magnetic
bead‐based
tailored
quantities
EVs
(0.5
1
µg
protein)
obtain
peptides
for
1908–2340
groups.
We
demonstrate
power
robustness
our
pipeline
in
obtaining
proteomes
granularity
different
sources
ascertain
known
This
underscores
capacity
optimised
workflow
capture
precise
comprehensive
EVs,
especially
ultra‐low
(sub‐nanogram),
an
important
challenge
field
where
in‐depth
information
essential.
Theranostics,
Год журнала:
2022,
Номер
12(15), С. 6548 - 6575
Опубликована: Янв. 1, 2022
Extracellular
vesicles,
especially
small
extracellular
vesicles
(sEVs)
are
now
accepted
as
important
messengers
in
cell-to-cell
communication
and
a
promising
drug
delivery
platform.They
involved
nearly
all
physiological
pathological
processes
disease
diagnosis
therapy.However,
their
heterogeneity
of
physicochemical
properties
functions
is
not
fully
understood,
which
hinders
further
clinical
applications.To
obtain
highly
bioactive
sEVs
with
both
high
yield
purity,
will
certainly
facilitate
future
study
application.This
review
informs
up-to-date
research
on
frequently-used
cutting-edge
technologies
isolation
makes
deep
comparison
analysis
different
methods,
including
advantages,
limitations
applications.Pending
questions
about
the
inherent
property
these
well
strategies
discussed.Additionally,
an
overview
applications
treatment,
some
on-going
trials,
also
reviewed.
Abstract
In
recent
decades,
research
on
Extracellular
Vesicles
(EVs)
has
gained
prominence
in
the
life
sciences
due
to
their
critical
roles
both
health
and
disease
states,
offering
promising
applications
diagnosis,
drug
delivery,
therapy.
However,
inherent
heterogeneity
complex
origins
pose
significant
challenges
preparation,
analysis,
subsequent
clinical
application.
This
review
is
structured
provide
an
overview
of
biogenesis,
composition,
various
sources
EVs,
thereby
laying
groundwork
for
a
detailed
discussion
contemporary
techniques
preparation
analysis.
Particular
focus
given
state‐of‐the‐art
technologies
that
employ
microfluidic
non‐microfluidic
platforms
EV
processing.
Furthermore,
this
discourse
extends
into
innovative
approaches
incorporate
artificial
intelligence
cutting‐edge
electrochemical
sensors,
with
particular
emphasis
single
proposes
current
outlines
prospective
avenues
future
research.
The
objective
motivate
researchers
innovate
expand
methods
analysis
fully
unlocking
biomedical
potential.
Abstract
Extracellular
vesicles
(EVs)
contain
various
regulatory
molecules
and
mediate
intercellular
communications.
Although
EVs
are
secreted
from
cell
types,
including
skeletal
muscle
cells,
present
in
the
blood,
their
identity
is
poorly
characterized
vivo,
limiting
identification
of
origin
blood.
Since
largest
organ
body,
it
could
substantially
contribute
to
circulating
as
source.
However,
due
lack
defined
markers
that
distinguish
muscle-derived
(SkM-EVs)
others,
whether
releases
vivo
how
much
SkM-EVs
account
for
plasma
remain
understood.
In
this
work,
we
perform
quantitative
proteomic
analyses
on
released
C2C12
cells
human
iPS
cell-derived
myocytes
identify
potential
marker
proteins
mark
SkM-EVs.
These
identified
apply
tracking
The
results
show
makes
only
a
subtle
contribution
source
both
control
exercise
conditions
mice.
On
other
hand,
demonstrate
concentrated
interstitium.
Furthermore,
interstitium
highly
enriched
with
muscle-specific
miRNAs
repress
expression
paired
box
transcription
factor
Pax7,
master
regulator
myogenesis.
Taken
together,
our
findings
confirm
previous
studies
showing
release
exosome-like
specific
protein
miRNA
profiles
suggest
mainly
play
role
within
microenvironment
where
they
accumulate.
Abstract
Extracellular
vesicles
(EVs)
are
extremely
versatile
naturally
occurring
membrane
particles
that
convey
complex
signals
between
cells.
EVs
of
different
cellular
sources
capable
inducing
striking
therapeutic
responses
in
neurological
disease
models.
Differently
from
pharmacological
compounds
act
by
modulating
defined
signalling
pathways,
EV-based
therapeutics
possess
multiple
abilities
via
a
variety
effectors,
thus
allowing
the
modulation
processes
may
have
very
potent
effects
on
brain
tissue
recovery.
When
applied
vivo
experimental
models
diseases,
revealed
remarkable
immune
responses,
cell
metabolism
and
neuronal
plasticity.
This
multimodal
neuroimmune
networks
profoundly
influences
highly
synergistic
context-dependent
way.
Ultimately,
EV-mediated
restoration
functions
helps
to
set
stage
for
With
this
review
we
first
outline
current
understanding
mechanisms
action
EVs,
describing
how
released
various
identify
their
targets
recipient
Then,
applicable
key
conditions
such
as
stroke,
sclerosis
neurodegenerative
diseases
presented.
Pathways
deserve
attention
specific
contexts
discussed.
We
subsequently
showcase
considerations
about
EV
biodistribution
delineate
genetic
engineering
strategies
aiming
at
enhancing
uptake
signalling.
By
sketching
broad
view
EV-orchestrated
plasticity
recovery,
finally
define
possible
future
clinical
applications
propose
necessary
information
be
provided
ahead
trials.
Our
goal
is
provide
steppingstone
can
used
critically
discuss
next
generation
diseases.
