Emergent
breast
tumor
resistance
and
microenvironment
(TME)
heterogeneity
can
lead
to
decreased
drug
delivery
efficacy,
resulting
in
therapeutic
failure.
Preclinical
molecular
imaging
is
a
crucial
tool
the
advancement
of
targeted
therapeutics
for
supporting
development
new
drugs
but
also
elucidate
factors
hampering
optimal
delivery.
However,
noninvasive
modalities
that
quantify
drug-target
engagement,
which
critical
actuation,
are
still
lacking.
We
have
demonstrated
utility
macroscopic
fluorescence
lifetime
Forster's
Resonance
Energy
Transfer
(MFLI
FRET)-based
optical
measure
labeled
trastuzumab
(TZM)-human
epidermal
growth
factor
receptor
(HER2)
binding
human
HER2+
cell
lines
xenograft
mice
models.
established
clinically
relevant
TZM
antibody
containing
Meditope
(MDT)
peptide
conjugated
near-infrared
(NIR)
dyelabeled
FRET
pairs,
retain
full
HER2
capability.
Herein,
we
demonstrate
measurements
using
MFLI
vivo
platform
ability
MDT-TZM
bind
living
xenografts.
AU565
xenografts
bearing
nude
were
injected
retro-orbitally
with
(NHS-conjugated)
or
MDTTZM
AlexaFluor700
(donor)
AlexaFluor750
(acceptor)
MFLIFRET
was
performed
24
h
48
post-injection.
Preliminary
data
suggest
shows
higher
uniform
consistent
signal
compared
TZM,
suggesting
increased
efficacy
TZM-MDT-HER2
binding.
Also
staggered
injections
donor
acceptor
may
be
quantifying
single
injections.
Biomedical Optics Express,
Год журнала:
2022,
Номер
13(9), С. 4637 - 4637
Опубликована: Июль 19, 2022
We
report
on
the
system
design
and
instrumental
characteristics
of
a
novel
time-domain
mesoscopic
fluorescence
molecular
tomography
(TD-MFMT)
for
multiplexed
imaging
in
turbid
media.
The
is
equipped
with
supercontinuum
pulsed
laser
broad
spectral
excitation,
based
high-density
descanned
raster
scanning
intensity-based
acquisition
2D
3D
augmented
high-dynamical
range
linear
time-resolved
single-photon
avalanche
diode
(SPAD)
array
lifetime
quantification.
system’s
spatio-temporal
its
sensitivity
specificity
controlled
experimental
settings.
Also,
phantom
study
undertaken
to
test
performance
image
deeply-seated
inclusions
tissue-like
In
addition,
ex
vivo
tumor
xenograft
performed
validate
applicability
biological
sample.
characterization
results
manifest
capability
sense
small
concentrations
(on
order
nanomolar)
while
quantifying
lifetimes
lifetime-based
parameters
at
high
resolution.
demonstrate
potential
perform
thanks
contrast
(at
millimeters
depth).
exhibits
prospect
TD-MFMT
resolve
intra-tumoral
heterogeneity
depth-dependent
manner.
Sensors and Actuators B Chemical,
Год журнала:
2022,
Номер
371, С. 132486 - 132486
Опубликована: Авг. 5, 2022
Optochemical
sensors
are
actively
used
in
cell
analysis,
however
existing
systems
have
limitations
with
respect
to
their
robustness
and
analytical
performance.
We
developed
advanced
multimodal
multi-parametric
solid-state
pH
for
analysis
based
on
hydrophobic
protonable
metal-free
porphyrins,
such
as
octaethylporphine
(OEP),
octaethylporphine-ketone
(OEPK),
fluorescent
indicators.
The
internally
referenced
ratiometric
intensity
nanosecond
lifetime-based
versions
of
the
were
also
multiplexed
O2
phosphorescent
PtOEP
dye.
optimised
key
parameters
sensor,
including:
dye
encapsulation
matrix,
type
concentration
proton
transfer
reagent,
measurement
range
pKa,
concentrations
cross-talk
sensor.
Subsequently,
sensor
coatings
deposited
common
substrates
(96-well
plates),
fine-tuned
operational
performance,
dual
O2/pH
sensing
functionally
ability
measure
Extracellular
Acidification
(ECAR)
Oxygen
Consumption
(OCR)
rates
biological
samples
containing
cells.
stable
calibrations,
convenient
spectral
characteristics
low
cytotoxicity,
demonstrated
cultured
cells
3D
spheroid
structures,
measuring
ECAR,
OCR
responses
stimulation.
These
pH/O2
well-suited
detailed
metabolic
studies
widely
available
laboratory
equipment.
Journal of Visualized Experiments,
Год журнала:
2022,
Номер
182
Опубликована: Апрель 6, 2022
Multicellular
spheroids
are
important
tools
for
studying
tissue
and
cancer
physiology
in
3D
frequently
used
engineering
as
assembling
units
biofabrication.
While
the
main
power
of
spheroid
model
is
mimicking
physical-chemical
gradients
at
microscale,
real
physiological
environment
(including
dynamics
metabolic
activity,
oxygenation,
cell
death,
proliferation)
inside
generally
ignored.
At
same
time,
effects
growth
medium
composition
formation
method
on
resulting
phenotype
well
documented.
Thus,
characterization
standardization
required
to
ensure
reproducibility
transparency
research
results.
The
analysis
average
oxygenation
value
O2
three
dimensions
(3D)
can
be
a
simple
universal
way
characterization,
pointing
their
overall
viability,
potential
recapitulate
vivo
microenvironment.
visualization
easily
combined
with
multiparametric
additional
parameters
(such
proliferation,
composition)
applied
continuous
monitoring
and/or
end-point
measurements.
loading
probe
performed
during
stage
compatible
various
protocols
generation.
protocol
includes
high-throughput
generation
introduced
red
near-infrared
emitting
ratiometric
fluorescent
nanosensors
description
multi-parameter
assessment
death
before
after
bioprinting.
experimental
examples
show
comparative
homo-
hetero-cellular
spheroid-based
bioprinted
constructs.
conventional
fluorescence
microscope
having
multiple
filters
light-emitting
diode
light
source.
Advanced Science,
Год журнала:
2024,
Номер
unknown
Опубликована: Ноя. 27, 2024
Abstract
Trastuzumab
(TZM)
is
a
monoclonal
antibody
that
targets
the
human
epidermal
growth
factor
receptor
2
(HER2)
and
clinically
used
for
treatment
of
HER2‐positive
breast
tumors.
However,
tumor
microenvironment
can
limit
access
TZM
to
HER2
across
whole
thereby
compromising
TZM's
therapeutic
efficacy.
An
imaging
methodology
non‐invasively
quantify
binding
TZM‐HER2,
which
required
action,
distribution
within
tumors
with
varying
microenvironments
much
needed.
Near‐infrared
(NIR)
fluorescence
lifetime
(FLI)
Forster
Resonance
Energy
Transfer
(FRET)
performed
measure
TZM‐HER2
binding,
using
in
vitro
microscopy
vivo
widefield
macroscopy,
overexpressing
ovarian
cancer
cells
xenografts,
respectively.
Immunohistochemistry
validate
results.
NIR
FLI
FRET
data
show
variations
intracellular
bound
AU565
tumor‐passaged
XTM
cell
lines
comparison
SKOV‐3
cells.
Macroscopy
(MFLI)
display
reduced
compared
tumors,
as
validated
by
ex
immunohistochemistry.
Moreover,
AU565/XTM
xenografts
different
amounts
distributions
TME
components,
such
collagen
vascularity.
Therefore,
these
results
suggest
are
refractory
delivery
due
their
disrupted
vasculature
increased
content.
The
study
demonstrates
powerful
analytical
tool
monitor
antibodydrugs
both
cultures
live
systems.
Especially,
MFLI
unique
modality
directly
target
engagement
potential
elucidate
role
drug
efficacy
intact
xenografts.
Emergent
breast
tumor
resistance
and
microenvironment
(TME)
heterogeneity
can
lead
to
decreased
drug
delivery
efficacy,
resulting
in
therapeutic
failure.
Preclinical
molecular
imaging
is
a
crucial
tool
the
advancement
of
targeted
therapeutics
for
supporting
development
new
drugs
but
also
elucidate
factors
hampering
optimal
delivery.
However,
noninvasive
modalities
that
quantify
drug-target
engagement,
which
critical
actuation,
are
still
lacking.
We
have
demonstrated
utility
macroscopic
fluorescence
lifetime
Forster's
Resonance
Energy
Transfer
(MFLI
FRET)-based
optical
measure
labeled
trastuzumab
(TZM)-human
epidermal
growth
factor
receptor
(HER2)
binding
human
HER2+
cell
lines
xenograft
mice
models.
established
clinically
relevant
TZM
antibody
containing
Meditope
(MDT)
peptide
conjugated
near-infrared
(NIR)
dyelabeled
FRET
pairs,
retain
full
HER2
capability.
Herein,
we
demonstrate
measurements
using
MFLI
vivo
platform
ability
MDT-TZM
bind
living
xenografts.
AU565
xenografts
bearing
nude
were
injected
retro-orbitally
with
(NHS-conjugated)
or
MDTTZM
AlexaFluor700
(donor)
AlexaFluor750
(acceptor)
MFLIFRET
was
performed
24
h
48
post-injection.
Preliminary
data
suggest
shows
higher
uniform
consistent
signal
compared
TZM,
suggesting
increased
efficacy
TZM-MDT-HER2
binding.
Also
staggered
injections
donor
acceptor
may
be
quantifying
single
injections.