bioRxiv (Cold Spring Harbor Laboratory),
Journal Year:
2024,
Volume and Issue:
unknown
Published: Dec. 22, 2024
Abstract
Lipid
nanoparticles
(LNPs)
are
the
most
advanced
delivery
system
currently
available
for
RNA
therapeutics.
Their
development
has
accelerated
since
success
of
Patisiran,
first
siRNA-LNP
therapeutic,
and
mRNA
vaccines
that
emerged
during
COVID-19
pandemic.
Designing
LNPs
with
specific
targeting,
high
potency,
minimal
side
effects
is
crucial
their
successful
clinical
use.
These
characteristics
have
been
improved
through
microfluidic
platforms,
which
enhanced
efficacy
uniformity
LNP
batches.
However,
our
understanding
how
composition
mixing
method
influences
structural,
biophysical,
biological
properties
resulting
particles
remains
limited,
hindering
LNPs.
Our
lack
structural
extends
from
physical
compositional
polydispersity
LNPs,
render
traditional
characterization
methods,
such
as
dynamic
light
scattering
(DLS),
unable
to
accurately
quantitate
physicochemical
In
this
study,
we
address
challenge
structurally
characterizing
polydisperse
formulations
using
emerging
solution-based
biophysical
methods
higher
resolution
provide
data
beyond
size
polydispersity.
techniques
include
sedimentation
velocity
analytical
ultracentrifugation
(SV-AUC),
field
flow
fractionation
followed
by
multi-angle
(FFF-MALS),
size-exclusion
chromatography
in-line
synchrotron
small-angle
X-ray
(SEC-SAXS).
Here,
show
intrinsic
in
size,
loading,
shape,
these
parameters
dependent
on
both
formulation
technique
lipid
composition.
Lastly,
demonstrate
can
be
employed
predict
transfection
three
models
examining
relationship
between
translation
characteristics.
We
envision
employing
will
essential
determining
structure-function
relationships,
facilitating
creation
new
design
rules
future
bioRxiv (Cold Spring Harbor Laboratory),
Journal Year:
2024,
Volume and Issue:
unknown
Published: Nov. 15, 2024
The
unprecedented
success
of
mRNA-lipid
nanoparticles
(LNPs)
has
highlighted
their
power
for
protein
expression,
but
the
hours-long
half-life
mRNA
severely
limits
use
in
chronic
diseases.
In
contrast,
DNA
LNPs
display
months-long
expression
and
genetically
encode
cell
type
specificity,
been
hindered
by
poor
(orders
magnitude
lower
than
LNPs).
To
overcome
this,
we
introduce
multi-stage
mixing
(MSM)
microfluidics
to
control
internal
structure
it
create
core-then-shell
(CTS)
structured
LNPs.
CTS
distinct
thermal
transitions
organization
compared
amorphous
conventional
improves
transfection
three
orders
ACS Bio & Med Chem Au,
Journal Year:
2024,
Volume and Issue:
5(1), P. 154 - 165
Published: Nov. 27, 2024
The
current
rapid
advancement
in
ribonucleic
acid
(RNA)
therapeutics
research
depends
on
innovations
drug
delivery,
especially
the
development
of
a
lipid-nanoparticle
(LNP)-based
system.
conventional
LNP
formulation
typically
contains
four
components,
including
an
ionizable
cationic
lipid,
phospholipid,
cholesterol
or
derivative,
and
poly(ethylene
glycol)
(PEG)-lipid,
with
each
contributing
to
formulation's
overall
stability
effectiveness.
Among
these
types
lipids,
phospholipid
component
is
often
known
provide
structural
support
for
nanoparticles
but
also
class
bioactive
molecules
strong
cell
signaling
potential.
This
study
explores
possibility
incorporating
some
structurally
related
phospholipids
as
fifth
four-component
assesses
impacts
such
approach
physicochemical
properties
biological
functions
mRNA
formulation.
We
screened
library
formulations
containing
7
different
at
molar
concentrations
5%,
15%
30%
addition
(base).
observed
differences
between
that
could
be
attributed
both
examined
used.
Cryo-EM
analysis
revealed
similarity
Base
other
formulations.
characterized
protein
expression
level
HeLa
cells
picked
up
distinct
cytokine
panel
signature
human
peripheral
blood
mononuclear
(hPBMCs).
Further
immunophenotyping
showed
most
were
transfected
CD4+
T
cells,
slightly
altered
cellular
tropism.
exploratory
illustrates
how
adding
can
used
modulate
function,
further
expanding
design
space
RNA
potentiating
LNPs
use
therapeutics.
bioRxiv (Cold Spring Harbor Laboratory),
Journal Year:
2024,
Volume and Issue:
unknown
Published: Dec. 22, 2024
Abstract
Lipid
nanoparticles
(LNPs)
are
the
most
advanced
delivery
system
currently
available
for
RNA
therapeutics.
Their
development
has
accelerated
since
success
of
Patisiran,
first
siRNA-LNP
therapeutic,
and
mRNA
vaccines
that
emerged
during
COVID-19
pandemic.
Designing
LNPs
with
specific
targeting,
high
potency,
minimal
side
effects
is
crucial
their
successful
clinical
use.
These
characteristics
have
been
improved
through
microfluidic
platforms,
which
enhanced
efficacy
uniformity
LNP
batches.
However,
our
understanding
how
composition
mixing
method
influences
structural,
biophysical,
biological
properties
resulting
particles
remains
limited,
hindering
LNPs.
Our
lack
structural
extends
from
physical
compositional
polydispersity
LNPs,
render
traditional
characterization
methods,
such
as
dynamic
light
scattering
(DLS),
unable
to
accurately
quantitate
physicochemical
In
this
study,
we
address
challenge
structurally
characterizing
polydisperse
formulations
using
emerging
solution-based
biophysical
methods
higher
resolution
provide
data
beyond
size
polydispersity.
techniques
include
sedimentation
velocity
analytical
ultracentrifugation
(SV-AUC),
field
flow
fractionation
followed
by
multi-angle
(FFF-MALS),
size-exclusion
chromatography
in-line
synchrotron
small-angle
X-ray
(SEC-SAXS).
Here,
show
intrinsic
in
size,
loading,
shape,
these
parameters
dependent
on
both
formulation
technique
lipid
composition.
Lastly,
demonstrate
can
be
employed
predict
transfection
three
models
examining
relationship
between
translation
characteristics.
We
envision
employing
will
essential
determining
structure-function
relationships,
facilitating
creation
new
design
rules
future
