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Construction and evaluation of a novel humanized HER2-specific chimeric receptor.
[severe combined immunodeficiency]
The
human
epidermal
growth
factor
receptor
2
(
HER
2
)
represents
one
of
the
most
studied
tumor
-associated
antigens
(
TAAs
)
for
cancer
immunotherapy
.
The
monoclonal
antibody
(
mAb
)
trastuzumab
has
improved
the
outcomes
of
patients
with
HER
2
+
breast
cancer
.
However
,
a
large
number
of
HER
2
+
tumors
are
not
responsive
to
,
or
become
resistant
to
,
trastuzumab-based
therapy
,
and
thus
more
effective
therapies
targeting
HER
2
are
needed
.
H
ER
2
-
specific
T
cells
were
generated
by
the
transfer
of
genes
that
encode
chimeric
antigen
receptor
(
CAR
)
.
Using
a
multistep
overlap
extension
PCR
method
,
we
constructed
a
novel
,
humanized
HER
2
CAR-containing
,
chA
21
single
-chain
variable
fragment
(
scFv
)
region
of
antigen-
specific
mAb
and
T
-
cell
intracellular
signaling
chains
made
up
of
CD
28
and
CD
3
ζ
.
An
interferon
γ
and
interleukin
2
enzyme-linked
immunosorbent
assay
and
a
chromium-
51
release
assay
were
used
to
evaluate
the
antitumor
immune
response
of
CAR
T
cells
in
coculture
with
tumor
cells
.
Furthermore
,
SKBR
3
tumor
-bearing
nonobese
diabetic
/
severe
combined
immunodeficiency
(
NOD
/
SCID
)
mice
were
treated
with
HER
2
CAR
T
cells
to
evaluate
antitumor
activity
.
Human
CD
3
+
T
cell
accumulation
in
tumor
xenograft
was
detected
by
immunohistochemistry
.
chA
21
-
28
z
CAR
was
successfully
constructed
,
and
both
CD
4
+
and
CD
8
+
T
cells
were
transduced
.
The
expanded
HER
2
CAR
T
cells
expressed
a
central
memory
phenotype
and
specifically
reacted
against
HER
2
+
tumor
cell
lines
.
Furthermore
,
the
SKBR
3
tumor
xenograft
model
revealed
that
HER
2
CAR
T
cells
significantly
inhibited
tumor
growth
in
vivo
.
Immunohistochemical
analysis
showed
robust
accumulation
of
human
CD
3
+
T
cells
in
regressing
SKBR
3
lesions
.
The
results
of
this
study
show
that
novel
chA
21
scFv-based
,
HER
2
-
specific
CAR
T
cells
not
only
recognized
and
killed
HER
2
+
breast
and
ovarian
cancer
cells
ex
vivo
but
also
induced
regression
of
experimental
breast
cancer
in
vivo
.
Our
data
support
further
exploration
of
the
HER
2
CAR
T
-
cell
therapy
for
HER
2
-
expressing
cancers
.
Diseases
Validation
Diseases presenting
"tumor cells"
symptom
alpha-thalassemia
carcinoma of the gallbladder
cholangiocarcinoma
cushing syndrome
dedifferentiated liposarcoma
dentin dysplasia
esophageal adenocarcinoma
esophageal carcinoma
esophageal squamous cell carcinoma
hodgkin lymphoma, classical
junctional epidermolysis bullosa
kindler syndrome
liposarcoma
lymphangioleiomyomatosis
pleomorphic liposarcoma
primary effusion lymphoma
severe combined immunodeficiency
triple a syndrome
von hippel-lindau disease
waldenström macroglobulinemia
well-differentiated liposarcoma
werner syndrome
wiskott-aldrich syndrome
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