Journal for ImmunoTherapy of Cancer,
Год журнала:
2022,
Номер
10(9), С. e005189 - e005189
Опубликована: Сен. 1, 2022
Background
There
is
an
increasing
demand
for
chimeric
antigen
receptor
(CAR)
T
cell
products
from
patients
and
care
givers.
Here,
we
established
automated
manufacturing
process
CAR
cells
on
the
CliniMACS
Prodigy
platform
that
scaled
to
provide
therapeutic
doses
achieves
gene-transfer
with
virus-free
Sleeping
Beauty
(SB)
transposition.
Methods
We
used
advanced
connected
electroporator
unit
performed
a
series
of
small-scale
development
large-scale
confirmation
runs
primary
human
cells.
Transposition
was
accomplished
minicircle
(MC)
DNA-encoded
SB100X
transposase
pT2
transposon
encoding
CD19
CAR.
Results
defined
bi-pulse
electroporation
shock
bi-directional
unidirectional
electric
field,
respectively,
permitted
efficient
MC
insertion
maintained
high
frequency
viable
In
three
large
scale
runs,
2E8
were
enriched
leukapheresis
product,
activated,
gene-engineered
expanded
yield
up
3.5E9
total
cells/1.4E9
CAR-modified
within
12
days
(CAR-modified
cells:
28.8%±12.3%).
The
resulting
product
contained
highly
pure
(97.3±1.6%)
balanced
CD4/CD8
ratio
central
memory
phenotype
(87.5%±10.4%).
copy
number
7.0,
9.4
6.8
in
#1–3,
gene
analyses
showed
expression
activation/exhaustion
markers.
conferred
potent
anti-lymphoma
reactivity
pre-clinical
models.
Notably,
operator
hands-on-time
substantially
reduced
compared
conventional
non-automated
campaigns.
Conclusions
report
first
transposon-based
ready
formal
validation
use
clinical
This
have
potential
facilitate
access
therapy
accelerate
scaled,
multiplexed
both
academic
industry
setting.
Science,
Год журнала:
2022,
Номер
378(6622), С. 853 - 858
Опубликована: Ноя. 24, 2022
Immune
cells
are
being
engineered
to
recognize
and
respond
disease
states,
acting
as
a
"living
drug"
when
transferred
into
patients.
Therapies
based
on
immune
now
clinical
reality,
with
multiple
T
cell
therapies
approved
for
treatment
of
hematologic
malignancies.
Ongoing
preclinical
studies
testing
diverse
strategies
modify
the
fate
function
applications
in
cancer,
infectious
disease,
beyond.
Here,
we
discuss
current
progress
treating
human
therapeutics,
emerging
engineering,
challenges
facing
field,
particular
emphasis
where
most
effort
has
been
applied
date.
Science Immunology,
Год журнала:
2023,
Номер
8(82)
Опубликована: Апрель 14, 2023
Chimeric
antigen
receptor
(CAR)
T
cells
have
achieved
true
clinical
success
in
treating
hematological
malignancy
patients,
laying
the
foundation
of
CAR
as
a
new
pillar
cancer
therapy.
Although
these
promising
effects
generated
strong
interest
expanding
treatment
to
solid
tumors,
reproducible
demonstration
efficacy
setting
tumors
has
remained
challenging
date.
Here,
we
review
how
metabolic
stress
and
signaling
tumor
microenvironment,
including
intrinsic
determinants
response
cell
therapy
extrinsic
obstacles,
restrict
treatment.
In
addition,
discuss
use
novel
approaches
target
rewire
programming
for
manufacturing.
Last,
summarize
strategies
that
aim
improve
adaptability
enhance
their
potency
mounting
antitumor
responses
survival
within
microenvironment.
Molecular Therapy — Methods & Clinical Development,
Год журнала:
2023,
Номер
31, С. 101139 - 101139
Опубликована: Окт. 18, 2023
Engineered
T
cells
expressing
chimeric
antigen
receptors
(CARs)
have
been
proven
as
efficacious
therapies
against
selected
hematological
malignancies.
However,
the
approved
CAR
cell
therapeutics
strictly
rely
on
viral
transduction,
a
time-
and
cost-intensive
procedure
with
possible
safety
issues.
Therefore,
direct
transfer
of
in
vitro
transcribed
CAR-mRNA
into
is
pursued
promising
strategy
for
engineering.
Electroporation
(EP)
currently
used
mRNA
delivery
method
generation
clinical
trials
but
achieving
only
poor
anti-tumor
responses.
Here,
lipid
nanoparticles
(LNPs)
were
examined
ex
vivo
compared
EP.
LNP-CAR
showed
significantly
prolonged
efficacy
comparison
EP-CAR
result
extended
persistence
expression,
attributed
to
different
mechanism
less
cytotoxicity
slower
proliferation.
Moreover,
expression
functionality
mRNA-LNP-derived
comparable
stably
transduced
exhausted.
These
results
show
that
LNPs
outperform
EP
underline
great
potential
mRNA-LNP
modification
next-generation
transient
approach
studies.
Frontiers in Immunology,
Год журнала:
2023,
Номер
14
Опубликована: Май 2, 2023
Advancements
in
chimeric
antigen
receptor
engineered
T-cell
(CAR-T)
therapy
have
revolutionized
treatment
for
several
cancer
types
over
the
past
decade.
Despite
this
success,
obstacles
including
high
price
tag,
manufacturing
complexity,
and
treatment-associated
toxicities
limited
broad
application
of
therapy.
Chimeric
natural
killer
cell
(CAR-NK)
offers
a
potential
opportunity
simpler
more
affordable
“off-the-shelf”
treatment,
likely
with
fewer
toxicities.
Unlike
CAR-T,
CAR-NK
therapies
are
still
early
development,
few
clinical
trials
yet
reported.
Given
challenges
experienced
through
development
CAR-T
therapies,
review
explores
what
lessons
we
can
apply
to
build
better
therapies.
In
particular,
explore
importance
optimizing
immunochemical
properties
CAR
construct,
understanding
factors
leading
product
persistence,
enhancing
trafficking
transferred
cells
tumor,
ensuring
metabolic
fitness
product,
strategies
avoid
tumor
escape
loss.
We
also
trogocytosis,
an
important
emerging
challenge
that
equally
applies
cells.
Finally,
discuss
how
these
limitations
already
being
addressed
future
directions
may
be
possible.
