harnessing the innate and adaptive immune system to eradicate … · harnessing the innate and...
Post on 27-Jul-2018
214 Views
Preview:
TRANSCRIPT
Harnessing the innate and adaptive immune system to eradicate treated and distant untreated solid tumorsSaul Kivimäe, Werner Rubas, Rhoneil Pena, Joseph McLaughlin, Marlene Hennessy, Yolanda Kirksey, Wildaliz Nieves, Myong Lee, Clive Law, Kavitha Bhasi, Phi Quach, Janet Cetz, John L. Langowski, Christie Fanton, Jose Zandro Aquino, Zhongxu Ren, Haiying Cai, BoLiang Deng, Wen Zhang, Neel K. Anand, Jennifer A. Riggs-Sauthier, Steve Doberstein, Jonathan Zalevsky
Nektar Therapeutics, San Francisco, CA
INTRODUCTION
RESULTS
RESULTS
• Tumor antigen release and T cell priming by antigen presenting cells is a critical �rst step for tumor growth inhibition by the adaptive immune system
• Toll-like-receptor (TLR) stimulation can induce differentiation of functional antigen presenting cells in the tumor environment and reduce immune suppression in tumors facilitating T cell priming
• Pharmacological induction of tumor antigen presentation combined with sustained in vivo expansion of tumor speci�c CD8 T cells can potentially increase diversity and numbers of tumor killing cytotoxic T cells enabling ef�cacious anti-tumor immune therapies
• Combination treatment with a novel intratumoral TLR7/8 targeting agent NKTR-262 and a systemic CD122-biased agonist NKTR-214 leads to synergistic activation of innate and adaptive anti-tumor immune response resulting in highly ef�cacious growth inhibition of treated and abscopal lesions in multiple preclinical mouse tumor models
− Intratumorally delivered NKTR-262 provides local sustained release of TLR7/8 agonist with minimal systemic exposure
− Systemic sustained CD122-biased IL-2 pathway activation by NKTR-214 expands and maintains tumor in�ltrating CD8+ T cell clones
Bilateral subcutaneous tumors model NKTR-262 abscopal effect:
Single-sided subcutaneous tumors:
Right �ank tumors in mice bearing subcutaneous bilateral CT26 tumors were intratumorally treated with NKTR-262 (0.1 µg or 10 µg) on Day 0. NKTR-214 (0.8 mg/kg) was administered IV on Day 4. Immune cells in blood and in both treated and abscopal tumors were analyzed by �ow cytometry on Day 1 (NKTR-262 single agent activity) and Day 7 (NKTR-262 + NKTR-214 combination activity). (*p<0.05 with bars indicating comparisons)
Right �ank tumors in mice bearing subcutaneous bilateral CT26 tumors were treated intratumorally with NKTR-262 (10 µg). NKTR-262 and released TLR7/8 agonist concentrations were measured from both treated and abscopal left �ank tumors and from plasma at indicated timepoints after treatment.
CT26 tumor bearing mice were treated intratumorally with 10 µg of NKTR-262. Cytokine levels in treated tumors and in plasma were measured at Tmax of each cytokine (2 hr and 6 hr after treatment).
TLR7 and type I interferon pathway gene expression induction after NKTR-262 dose response measured by qRT-PCR relative to vehicle in CT26 subcutaneous tumors.(*p<0.05 relative to vehicle at Tmax)
A single NKTR-262 dose (20 µg) combined with NKTR-214 (0.8 mg/kg, i.v., q9d x 3 doses) inhibits tumor growth in several syngeneic mouse tumor models. Note: only the right �ank tumor was treated with NKTR-262 in bilateral models to assess abscopal ef�cacy in the untreated left �ank tumors (*p<0.05 relative to vehicle).
NKTR-262 and NKTR-214 engage non-overlapping immune mechanisms enhancing antigen presentation and anti-tumor T cell response
Complete anti-tumor immune response cascade induction by NKTR-262 and NKTR-214 − combination treatment optimally couples locally initiated intratumoral innate immune activation with systemic expansion and in�ltration of cytotoxic tumor reactive CD8 T cells
NKTR-262 + NKTR-214 combination treatment leads to relative decrease of immune suppressive cells in tumors
The injected tumors exhibit extended exposure to TLR7/8 agonist with minimal systemic exposure after local NKTR-262 administration
Rapid and transient activation of TLR7 pathway and type I interferon response genes in NKTR-262 treated tumors
Effective tumor growth inhibition by combining NKTR-262 with NKTR-214 in multiple preclinical syngeneic tumor models
Poster #P275: Society for Immunotherapy of Cancer 2017 Annual Meeting
Vehicle
NKTR-262 + NKTR-214
NKTR-262
NKTR-214
Vehicle
NKTR-262 + NKTR-214
NKTR-262NKTR-214
Treated tumors:
Abscopal tumors:
0 10 20 300
300
600
900
1200
EMT6 (Bilateral)Mammary Carcinoma
Days after treatment start
Tum
or
Vo
lum
e (m
m3
± S
EM
)
100% CR
CT26 (Bilateral)Colon Carcinoma
0 10 20 300
300
600
900
1200
Days after treatment start
Tum
or
Vo
lum
e (m
m3
± S
EM
)
100% CR
0
20000
40000
60000
80000
Tumor Dendritic CellsActivation (MHCII MFI)
Mea
n F
luo
resc
ence
Inte
nsi
ty
*
*
Day 1 Day 7
% o
f L
euko
cyte
s
0
20
40
60
Tumor Macrophages
**
Day 1 Day 70
10
20
30
40
Tumor CD4 Treg(CD25+ FoxP3+)
% o
f T
lym
ph
ocy
tes
*
Day 1 Day 70
1
2
3
4
5
Tumor Monocytes
% o
f L
ive
Cel
ls
*
Day 1 Day 7
Vehicle treated
Abscopal
0.1 µg NKTR-262
Abscopal
10 µg NKTR-262
Abscopal
+ NKTR-214
Day 1 Day 70
20
40
60
80
100
Lymph Node TargetedTumor Dendritic Cells
(CCR7+)
% o
f D
end
riti
c C
ells
*
0
20
40
60
Antigen ExperiencedTumor CD8 T Cells
(CD44hi)
% o
f C
yto
toxi
c T
Lym
ph
ocy
tes
*
Day 1 Day 7
Tumor CD8 T Cells
0
5
10
15
20
25
% o
f L
ive
Cel
ls
*
*
*
Day 1 Day 7
0
25
50
75
100
Tumor CD8/Treg
TIL
Rat
io
*
*
Day 1 Day 70
20
40
60
80
100
Checkpoint ReceptorsTumor CD8 T Cells
(PD-1+ CTLA-4+)
% o
f C
D8+
T L
ymp
ho
cyte
s *
Day 1 Day 7
Day 1 Day 70
20
40
60
80
100
Tumor Cell Viability
% o
f To
tal C
ells
* *
Day 1 Day 70
2
4
6
8
10
Tumor Neutrophils
% o
f L
ive
Cel
ls
*
*
Day 1 Day 70
20
40
60
80
100
Tumor Dendritic CellsActivation (CD86+)
% o
f D
end
riti
c C
ells
*
NKTR-262 and Released TLR7/8 Agonist Concentration in:
Treated Tumor
Time (hr)0 24 48 72 96 120 144
1
10
100
1000
10000
100000
Tum
or C
once
ntra
tion
(pm
ol/g
)
NKTR-262
TLR7/8 agonist
Plasma
1
10
100
1000
10000
100000
0 24 48 72 96 120 144
Time (hr)
Plas
ma
Conc
entr
atio
n (p
mol
/mL)
Abscopal Tumor
1
10
100
1000
10000
100000
0 24 48 72 96 120 144
Time (hr)
Tum
or C
once
ntra
tion
(pm
ol/g
)
2
0
20
40
60
80 Il12b
Fold
-ch
ang
e
6 24 48
20
10
20
30 Mx1
Hours post-dose
Fold
-ch
ang
e
6 24 482
0
2
4
6 Irf7
Hours post-dose6 24 48
Fold
-ch
ang
e
2
0
10
20
30
40
50Cxcl10
Hours post-dose Hours post-dose
Fold
-ch
ang
e
6 24 48
NKTR-262 (100 µg)NKTR-262 (10 µg)
Vehicle
NKTR-262 (1 µg)NKTR-262 (0.1 µg)NKTR-262 (0.01 µg)
*
Regulatory T cells are expanded selectively in peripheral blood
Intratumoral NKTR-262 administration leads to high proin�ammatory cytokines at treated tumor but reduced exposure in plasma
Day 1 Day 70.0
0.5
1.0
1.5
2.0
Blood CD4 Treg(CD25+ FoxP3+)
% o
f Leu
kocy
tes
*
Day 1 Day 70
5000
10000
15000
Blood CD4 Treg activation(CD25 MFI)
Mea
n Fl
uore
scen
ce In
tens
ity
*
Day 1 Day 70
2
4
6
8
Blood CD8/Treg
CD8/
Treg
Rat
io Vehicle
0.1 µg NKTR-262 + NKTR-214
10 µg NKTR-262 + NKTR-214
*
*
CONCLUSIONS• Intratumoral NKTR-262 treatment simultaneously induces tumor antigen release
and activation of antigen presenting cells
• NKTR-262 and NKTR-214 combination treatment relieves immune suppression selectively in tumor environment facilitating CD8 T cell activity
• NKTR-262 design enables TLR7/8 agonist release and induction of proin�ammatory cytokines at the injection site reducing systemic exposure
• NKTR-262 and NKTR-214 combination treatment optimally couples localized innate immune activation to systemic CD8 T cell expansion enhancing cytotoxic T cell in�ltration and activity in tumor lesions
NKTR-262Local i.t. dose
NKTR-214Systemic i.v.
TreatedtumorAbscopal
tumor
VehicleNKTR-262 + NKTR-214
0 10 20 300
250
500
750
1000
H22Hepatocarcinoma
*
Days after treatment start
Tum
or
Vo
lum
e (m
m3
± S
EM
)
0 5 10 15
WEHI-164Fibrosarcoma
*0
250
500
750
1000
Days after treatment start
Tum
or
Vo
lum
e (m
m3
± S
EM
)
0 5 10 150
500
1000
1500
2000
2500
3000
RM-1 − ProstateAdenocarcinoma
*
Days after treatment start
Tum
or
Vo
lum
e (m
m3
± S
EM
)
0 10 20 300
500
1000
1500
2000
2500
JC − MammaryAdenocarcinoma
*
Days after treatment start
Tum
or
Vo
lum
e (m
m3
± S
EM
)
Plasma and Tumor Cytokine Levels6 hr post dose
Co
nce
ntr
atio
n (p
g/m
L)
PlasmaTumor
IL-6
KC
/GR
O
TNFα
IL-1β
IFNγ
IL-5
IL-2
0
20
40
60
500
1000
1500
2000
100IL
-6
KC
/GR
O
TNFα
IL-1β
IL-5
IFNα
IFNγ
IL-2
IL-1
2p70
0
100
200
300
400
500
2000
4000
6000
Plasma and Tumor Cytokine Levels2 hr post dose
Co
nce
ntr
atio
n (p
g/m
L)
top related