NSCLC (adeno) HNSC BRCA COAD TNB KIRC KIRP...Preclinical Characterization of AB154, a Humanized...
Transcript of NSCLC (adeno) HNSC BRCA COAD TNB KIRC KIRP...Preclinical Characterization of AB154, a Humanized...
Preclinical Characterization of AB154, a Humanized α-TIGIT Antibody, For
Use in Combination Therapies
Anderson AE1, Becker A1, Yin F1, Singh H1, Zhao X1, Seitz L1, Udyavar A1, Stanton R2, Walker NPC1, Tan JBL1
1 Arcus Biosciences, Inc., Hayward, CA2 Stanton Biosciences, Newbury Park, CA
IntroductionTIGIT (T-cell immunoreceptor with Ig and ITIM domains) is an
inhibitory receptor expressed on NK and CD8+ T cells, as well as
immunosuppressive Tregs. TIGIT and DNAM-1 (CD226) are paired
receptors which compete for shared ligands CD155 (PVR) and
CD112 (Nectin-2). As malignancies progress, high TIGIT expression
often occurs alongside the up-regulation of other immune checkpoint
proteins and markers of T cell exhaustion such as PD-1. Shifting
CD155/CD112 from an unfavorable binding with immunosuppressive
TIGIT towards the more productive DNAM-1 interaction promotes T
cell and NK cell activation. We have developed AB154 to inhibit
and modulate the tumor microenvironment towards a more effective
anti-cancer response.
Figure 2. TIGIT and PD-1 expression were derived from RNAseq in
The Cancer Genome Atlas (TCGA) database. Numbers indicate log2
transformed expression of counts per million. Tumors with high
expression of both PD-1 and TIGIT (red), high TIGIT expression but
low PD-1 (blue) and lower levels of both TIGIT and PD-1 (black) are
shown. (B.) TIGIT and PD-1 expression on select tumor types
compared to solid normal tissue. *p ≤ 0.05. **p ≤ 0.01. ***p ≤ 0.001.
****p ≤ 0.0001.
PD-1 Expression Strongly Correlates
with TIGIT in Tumor Infiltrating T cells
Similar AB154 Potency on Peripheral
Blood Lymphocytes Isolated from
Healthy Donors and NSCLC Patients
AB154 Binding to Human TIGIT Blocks
Interaction with CD155
Conclusions
Figure 3. Tumor infiltrating lymphocytes (TILs) analyzed from
dissociated tumor samples revealed a strong correlation between
PD-1 and TIGIT expression on the surface of intratumoral T cells. (A.)
TIGIT is highly expressed on CD8+ and CD4+ CD25+ tumor infiltrating
T cells in different cancers. Right panel shows representative TIGIT
expression levels. (B.) Immunophenotyping of tumor infiltrating T cells
highlights significant TIGIT and PD-1 double positive cells.
Representative data from NSCLC shown.
B.
A.
Figure 4. (A.) In a CHO cell line over-expressing human TIGIT,
AB154 binds with sub-nanomolar affinity (0.35 nM). (B.) Binding of
soluble CD155-Fc to TIGIT was abrogated in the presence of AB154
with an IC50 of 0.69 nM.
Inhibition of TIGIT and PD-1 / PD-L1
Increases IFNγ Production in DC-MLR
AB• 154 is a humanized monoclonal antibody that potently
inhibits binding of human TIGIT to CD155.
TIGIT• and PD-1 expression are correlated in many tumor
types and are often co-expressed on tumor infiltrating
lymphocytes (TILs).
In• combination, AB154 enhances T cell activation relative to
α-PD-1 (AB122) or α-PD-L1 alone in MLR settings.
AB• 154 has sub-nanomolar potency on peripheral blood
lymphocytes derived from both healthy donors and NSCLC
patients.
Flow• cytometry will be used to monitor receptor occupancy
on CD8+ and CD4+ T cells, regulatory T cells, NK cells, and
NKT cells in Phase 1 dose escalation studies for AB154
monotherapy and AB154 / AB122 combination therapy.
Figure 6. Whole blood was obtained from healthy donors (n=3) and
non-small cell lung carcinoma (NSCLC) patients and assessed by
flow cytometry. (A.) Patients and healthy donors had similar
abundance of total lymphocytes, including CD8+ and CD8- T cells, NK
cells and NKT cells. TIGIT expression was also consistent in these
populations, as measured using saturating levels of a commercially
available α-TIGIT antibody. (B.) Fluorophore-conjugated AB154 was
used to directly determine binding affinity in whole blood, with
equipotency observed on lymphocytes isolated from healthy donors
and cancer patients.
Figure 5. Combinatorial settings were assessed using mixed
lymphocyte reactions (MLRs) consisting of CD4+ T cells and
monocytic DC cultures. Inhibition of TIGIT in combination with an (A.)
α-PD-L1 or (B.) α-PD-1 antibody (AB122) resulted in increased IFN-g
production relative to monotherapy. *p ≤ 0.05. **p ≤ 0.01.
Figure 7. (A.) TIGIT receptor occupancy was determined using
saturating levels (>EC90) of a commercially-available human α-TIGIT
antibody that binds competitively with AB154. Therefore, the apparent
IC50, as measured in this assay, is higher than the binding affinity of
AB154 shown in Figure 6B. (B.) In human whole blood, ex vivo
addition of AB154 achieved complete inhibition of TIGIT. Similar
receptor occupancy was observed between cancer patient and
healthy donor blood samples at all AB154 spike-in concentrations.
A Robust Assay to Measure Target
Engagement by AB154
TCGA Analysis of TIGIT and PD-1
Expression on Human Tumors
Figure 1. TIGIT binds to CD155 and results in decreased activation
of the TIGIT-expressing immune cells. AB154 blockade of TIGIT
allows CD155 to bind DNAM-1, favoring T cell and NK cell activation.
0
2 0
4 0
6 0
8 0
C D 4+
C D 2 5+
Pe
rc
en
t
T IG IT+
P D -1-
T IG IT+
P D -1+
T IG IT-
P D -1+
T IG IT-
P D -1-
0
1 0
2 0
3 0
4 0
5 0
C D 4+
C D 2 5-
Pe
rc
en
t
T IG IT+
P D -1-
T IG IT+
P D -1+
T IG IT-
P D -1+
T IG IT-
P D -1-
0
2 0
4 0
6 0
8 0
C D 8+
T C e lls
Pe
rc
en
t
T IG IT+
P D -1-
T IG IT+
P D -1+
T IG IT-
P D -1+
T IG IT-
P D -1-
Expression Level
+/-
++++
Cancer Cell type Cell Surface Markers TIGIT
TCRb+ CD4+ CD25- +/-
TCRb+ CD4+ CD25+ ++++
TCRb+ CD8+ ++++
CD11b+ CD14+ HLA-DR+ -
CD11b- CD14- HLA-DR+ -
TCRb+ CD4+ CD25- +/-
TCRb+ CD4+ CD25+ ++++
TCRb+ CD8+ ++++
CD11b+ CD14+ HLA-DR+ -
CD11b- CD14- HLA-DR+-
TCRb+ CD4+ CD25- +/-
TCRb+ CD4+ CD25+ ++++
TCRb+ CD8+++++
CD11b+ CD14+ HLA-DR+ -
CD11b- CD14- HLA-DR+-
TCRb+ CD4+ CD25- +/-
TCRb+ CD4+ CD25+ ++++
TCRb+ CD8+++++
CD11b+ CD14+ HLA-DR+ -
CD11b- CD14- HLA-DR+ -
SKCM
CRC
NSCLC
KIRC
T cells
Antigen Presenting Cells
T cells
Antigen Presenting Cells
T cells
Antigen Presenting Cells
T cells
Antigen Presenting Cells
0
1 0 0 0
2 0 0 0
3 0 0 0
IFN
-g (
pg
/ml)
A B 1 5 4 -P D -L 1 A B 1 5 4 +
-P D -L 1
Ig G 1
* *
A124
0
1 0 0 0
2 0 0 0
3 0 0 0
IFN
-g (
pg
/mL
)
*
Ig G 4 Ig G 1 Ig G 4 +
Ig G 1
A B 1 2 2 A B 1 2 2 +
A B 1 5 4
T cell and NK cell
Activation
CD155
TIGIT
DNAM
T cell and NK cell
Activation
CD155
TIGIT
DNAM
AB154
-2 0
0
2 0
4 0
6 0
8 0
1 0 0
L y m p h o c y t e P o p u la t io n s
Pe
rc
en
t
H e a lth y
N S C L C
C D 8+
T c e lls(T c e lls % )
C D 8-
T c e lls(T c e lls % )
N K T( ly m p h o c y t e s % )
N K( ly m p h o c y t e s % )
T o ta l
L y m p h o c y te s( s in g le t s % )
T c e lls( ly m p h o c y t e s % )
0
2 0
4 0
6 0
8 0
1 0 0
T IG IT + (% )
TIG
IT+
%
H e a lth y
N S C L C
C D 8+
T c e lls
C D 8-
T c e lls
N K T N KT o ta l
L y m p h o c y te s
T c e lls
A.
B.
B.
A.
Healthy Donors (n=3) NSCLC Patients (n=3)
0 .0 1 0 .1 1 1 0 1 0 0
0
1 0
2 0
3 0
4 0
A B 1 5 4 -A F 6 4 7 (n M )T
IGIT
+ %
(To
tal
Ly
mp
ho
cy
tes
)
M e a n E C 50 = 0 .1 3 n M
0 .0 1 0 .1 1 1 0 1 0 0
0
1 0
2 0
3 0
4 0
A B 1 5 4 -A F 6 4 7 (n M )
TIG
IT+
%
(To
tal
Ly
mp
ho
cy
tes
)
M e a n E C 50 = 0 .1 4 n M
0 .1 n M 1 n M 1 0 n M 1 0 0 n M
0
5 0
1 0 0
L y m p h o c y t e R e c e p t o r O c c u p a n c y
A B 1 5 4 S p ik e -In C o n c e n tra tio n s
Re
ce
pto
r
Oc
cu
pa
nc
y (
%)
N S C L C
H e a lth y
A. B.
A.
B.
A. B.
0 .0 1 0 .1 1 1 0 1 0 0
0
1 0 0 0 0 0
2 0 0 0 0 0
3 0 0 0 0 0
4 0 0 0 0 0
A B 1 5 4 (n M )
MF
I
h T IG IT
E C 50 = 0 .3 5 n M
-2 0 2 4
-2
-1
0
1
2
M e d ia n T IG IT C P M (L O G 2 )
Me
dia
n P
D-1
CP
M (
LO
G2
)
B L C A
B R C A
C E S C
C O A D
E R / P R + B R C A
E S C A
G B M
H E R 2 + B R C A
H N S C
K I R C
K I R PL I H C
N S C L C (a d e n o )
L U S C
O V
P A A D
P R A D
R E A D
S A R C
S K C M
S TA D TN B CU C E C
U C S
R2 = 0 .6 5 , p < 0 .0 0 0 1
0 .0 1 0 .1 1 1 0 1 0 0
0
2 0 0
4 0 0
6 0 0
8 0 0
1 0 0 0
A B 1 5 4 (n M )
MF
I
IC 50 = 0 .6 9 n M
AB154
Human
TIGIT
-1 0
-5
0
5
1 0
G B M
CP
M
* n s
P D -1 T IG IT-1 0
-5
0
5
1 0
K IR P
CP
M
* * *
P D -1 T IG IT-1 0
-5
0
5
1 0
B R C A
CP
M
P D -1 T IG IT
* * * * * * *
-1 0
-5
0
5
1 0
C O A D
CP
M
n s * *
P D -1 T IG IT
-1 0
-5
0
5
1 0
H N S C
CP
M
n s * * * *
P D -1 T IG IT-1 0
-5
0
5
1 0
K IR C
CP
M
* * * * * * * *
P D -1 T IG IT-1 0
-5
0
5
1 0
T N B C
CP
M
* * * * *
P D -1 T IG IT
-1 0
-5
0
5
1 0
N S C L C (a d e n o )
CP
M
* * * * * * * *
P D -1 T IG IT
S o lid N o rm a l T is s u e T u m o r