Harnessing the
Power of the
Host Immune
System to Fight
Cancer
Robert Kastelein
Immuno-Oncology Discovery
Texas FreshAIR Conference 2014
Houston October 23-24
Immunological Homeostasis
Activation Signal
Inhibitory Signal
T cell receptor
B cell receptor
NCR
Immunity
CD20
FcR
Homeostasis
Costimulatory Signal
2
CD28
CD40
Adhesion molecules
Cytokines
GITR
TIM1
HVEM
CD27
CTLA-4
PD-1
FcR
KIRs
Cytokines
TGFb
CD200R
TIM3
LAG3
Tolerance
Joe Phillips, Lewis Lanier
Role of PD-1 in Chronic Viral Infection
3
PD-1 is Important for Immune Tolerance
• PD-1 interacts with its ligands (PD-L1 and PD-L2) to inhibit
activation of T lymphocytes
• Cancers can ‘hijack’ the PD-1 pathway
• PD-1 blockade may reactivate anti-tumor immunity
*
MK-3475 Is a Potent Antagonistic
Antibody Against PD-1
• First-in-human study of MK-3475 in melanoma (Hamid et al, NEJM 369:134)
• Expanded study platform – reported activity in NSCLC, H&N cancer, bladder
and gastric cancer
• A signal detection study is exploring activity in 20 different tumor types
selected on the basis of PD-L1 expression
• Sep 4, 2014 – U.S approval of Keytruda for treatment of patients with
advanced or unresectable melanoma no longer responding to other treatments
Best Overall Response in Melanoma Patients
Treated with Pembrolizumab (RECIST 1.1)
ORR 34% by RECIST 1.1; 88% of responses ongoing
6
Kaplan Meier Estimate of PFS per
RECIST 1.1
69% OS at 12 months; median OS not reached
7
Immunotherapy in Oncology: Can we Alter the
Survival Curve by Combining Therapies?
Control
Standard or other therapy
Checkpoint blockade
Combination with standard
of care or other IMRs
Placing Immune Interventions in the
Context of Tumor Cell Recognition
Chemokines &
homing receptor
modulators
Cancer vaccines &
adjuvants
Cytotoxic agents
Adapted from Chen and Mellman, Immunity 39: 1 (2013)
Adoptive T cell
therapies
Immune checkpoint
targets and other
immunomodulators
Targeting Immunomodulators (IMRs):
The Opportunity and the Challenge
• By investigating how antiPD-1 works we will be
able to develop
meaningful combination
strategies
Melero I, et al. Clin Cancer Res. 2013;19:997-1008.
©2013 by American Association for Cancer Research.
Leveraging Preclinical Data to Inform IMR
Selection
Mouse Syngeneic
Tumor Models
•
•
•
•
•
11
Responder vs. non-responder
signature at baseline
Post-treatment signatures in
responder vs. non-responder
Flow cytometry of TILs, markers,
IMRs
IHC cell markers, IMRs
Gene expression in tumors
Human Clinical
Response with
MK-3475
Human Tumor
Analysis
Anti-mouse PD1 Surrogate is Efficacious in
Multiple, but not all, Syngeneic Tumor Models
Subcutaneous MC38
16 days post single injection of
DX-400
Subcutaneous CT-26
single injection of DX-400
ns
Tumor Volume (mm3)
2000
**
***
1500
1000
500
12
X40
0
D
X40
0
10
m
g/
K
g
D
g/
K
g
m
5
m
1
10
m
g/
K
g
g/
K
g
D
X40
0
Is
ot
yp
e
0
Initial tumor volumes at start ~100mm3
* Indicates statistical significance
PD1 and PDL1 are Upregulated Following
Anti-PD1 Treatment
DX400 0.3mpk
PDL1
DX400 0.3mpk
GLIOMA-26
Tumor Volume (mm3)
3000
2000
DX400 1.25mpk
DX400 1.25mpk
1000
0
0
5
10
15
20
Days post treatment
Isotype (0/5 CR)
0.3 mg/Kg muDX400 (0/5 CR)
0.6 mg/Kg muDX400 (0/5 CR)
1.25 mg/Kg muDX400 (2/5 CR)
2.5 mg/Kg muDX400 (4/5 CR)
5 mg/Kg muDX400 (4/5 CR)
DX400
5mpk
DX400
5mpk
PD1
Control mAb
Anti-PD-1 mAb induced Expansion of Mouse
Tumor-associated CD3 and CD8b+ T Cells
CD8
CD3
CD8
Anti-PD-1 mAb
CD3
14
MC38 mouse syngeneic tumor model
4 days post anti-PD-1 mAb treatment (DX400; at 5 mg/kg)
tumors were analyzed (pink – antigen; blue – nuclei)
4 days
post anti-PD-1
treatment
Anti-PD-1 Treatment Elicits Genes Associated
with Immune Response (preclinical models)
ene Expression
in aPD-1 treated mice
T umor expression
Blood Expression Ln Expression
Blood
LN
TUMOR
Cell Marker
Cell Marker
Tumor
Biology
Anti-Tumor
Immune Response
Chemokines
and Cell
Chemokines
and Cell
Activation/Recruitment
Activation/Recruitment
M2Immuno
and Immuno-Regulatory
M2 and
Regulatory
Profiling Mouse Models:
Blood ≠ LN ≠ Tumor
I
24h
pfd
I
4d
pfd
I
24h
psd
I
4d
psd
I
24h
pfd
I
4d
pfd
I
24h
psd
I
4d
psd
I
24h
pfd
I
4d
pfd
I
24h
psd
I
4d
psd
Intra-tumoral Clonal T-Cell Expansion
Following Anti-PD-1 Therapy
mDX400
0.35
Isotype control
6000
4000
2000
0
Naïve
Day 4
Day 9
TCR repertoire clonality,
tumor
# of T cells in tumor
tissue, tumor
8000
mDX400
Isotype control
0.3
0.25
0.2
0.15
Naïve
Time
Day 4
Day 9
Time
• MC38-tumor bearing mice were treated with 5 mg/kg anti-PD-1. Tumors were
harvested at 4 and 9 days following a single mAb dose.
– Increased intratumoral T cells at 4 and 9 days post-treatment
– T cell infiltrate is more clonal by 9 days post-treatment
16
Next-generation sequencing and data analyses were performed by Adaptive Biotechnologies.
Glucocorticoid-Induced TNFR-Related protein (GITR)
Biology and Target Rationale
17
Anti-GITR Treatment Specifically Targets
‘Intra-Tumor’ Tregs
Innoculate
MC38
Analyze TILs and draining LN
Anti-mGITR
or isotype
Kinetics of aGITR tx
response
Day
-8
0
1
4
12
T IL T r e g
T IL s
****
100
****
M C 3 8 is o ty p e
2000
M C 3 8 a G IT R
1500
1000
500
C D 8 :T re g ra tio
M C 3 8 is o ty p e
80
M C 3 8 a G IT R
60
40
20
0
2
1
y
a
y
a
D
D
D
a
D
y
a
1
y
2
4
y
a
D
D
a
y
1
1
0
4
T re g / m g tu m o r
2500
+
DLN
T c e lls
10
*
C D 8 :T re g ra tio
40000
20000
6000
4000
2000
8
6
4
2
1
2
4
y
y
a
D
D
a
D
y
a
a
y
2
1
4
y
a
D
D
a
y
1
18
1
0
0
D
CD8
+
T c e ll / m g t u m o r
T IL C D 8
Kinetics of Anti-GITR Treatment Response in
Tumor Micro-Environment
CTL
T cells /mg tumor
Treg
0
1
4
Days post anti-GITR treatment
19
12
Treatment with human GITR Agonist mAb Results in DoseDependent Reduction of MLR Induced Regulatory T cells
TREGs were induced by stimulating human PBMC with allogeneic dendritic cells for 7 days in the
presence of indicated concentrations of anti-human GITR.
***
***
**
40
30
20
g
0
0
2
1
G
IT
R
R
IT
G
IT
G
/m
/m
g
1
R
.1
0
R
IT
G
g
/m
l
/m
g
/m
g
u
0
2
e
p
ty
is
o
l
10
l
% C D 4 + /C D 2 5 + /F o x P 3 +
50
Treatment with anti-GITR agonist results in significant
reduction of FoxP3 TREGs
20
Cross-regulation of PD-1 and GITR in vivo
Upregulation of PD-1 and PD-L1 mRNA in
tumor after anti-GITR treatment
PD-1
PD-L1
Upregulation of GITR
mRNA in tumor after
anti-PD-1 treatment
Differential response
Min(normalized values)
GITR
d1 d4 d1 d4
d1 d4 d8 d12
Tumor
Anti-GITR
isotype control
d1 d4 d8 d12
Tumor
Anti-GITR single dose
isotype
anti-PD-1
Tumor
Anti-PD-1 single dose
Anti-PD-1 + Anti-GITR Combination Efficacy
Large MC38 tumors
Mean Tumor Volum (mm3)
(300mm3 starting volume)
3000
2/10 CR
Isotype ctrl d0,7,14
anti-GITR d0,7,14,21
anti-PD1 d0,7,14,21
10/10 CR
(anti-GITR+anti-PD1) d0,7
0/10 CR
2/10 CR
2000
1000
0
0
7
14
21
28
Day post anti-GITR and/or anti-PD1
All doses 5 mg/kg
●
●
●
●
22
Large established MC38 tumors used
Synergistic combination anti-tumor efficacy observed
Combination efficacy observed in additional models
All mAbs dosed at 5mg/kg
Exploiting the Patient’s Immune System for Optimal
Anti-Tumor Activity
Pushing the accelerator
•
•
•
•
•
•
•
cancer ‘vaccines’
adjuvants (e.g. BCG, TLRs)
tumor-specific antigens
adoptive immunotherapy
cytokines (IL-2, IL-12, IFNa)
cellular immunotherapy
agonists to immunostim.
IMRs – anti-GITR
Releasing the brakes

• anti-CTLA-4
• anti-PD-1

• antagonists to additional
inhibitory IMRs
Are both approaches needed for optimal efficacy?
23
Acknowledgements
• MK-3475 team and Anti-GITR team
• Discovery Team (Palo Alto & Boston) especially:
– Amy Beebe
– Smita Mauze
– Renu Jain
– Ashley Mahne
– Terri Mcclanahan
– Svetlana Sadekova
– Doug Wilson
– Debbie Law
24