Overcoming PD-1 targeting antibody resistance using combination strategies
#1681A
J.-F. MIRJOLET1, M. RAMELET1, D. FRANCE1, X. TIZON1, N. JEANRAY1
1 Oncodesign,
Dijon (France)
For more information: [email protected]
EMT6
PD-L1
PD-L2
B7-H3
B7-H4
?
?
CD200R
TIM3
BTLA
A2aR
CD200
Gal9
(HVEM) TNFSF14
Ade
(CD80) B7-1
(CD86) B7-2
Co-inhibitory
receptors
CTLA-4
R
E
S
U
L
T
S
Non-responders - Responders
Responders
Non-Responders
Comparison EMT6 & 4T1
EMT6: R - NR
4T1: R - NR
No NR
CD28
B7RP1
Co-stimulatory
receptors
ICOS
CD137L
CD137
OX40L
CD70
GITR-L
OX40
CD27
GITR
T/C
(D22)
Median TV
(mm3 at D22)
PD-1 mAb
76
470
CD137 mAb
58
360
GITR mAb
61
375
OX40 agonist mAb
20
126
Tim3 mAb
85
524
PD-1 + Anti-CD137 mAbs
47
292
PD-1 + Anti-GITR mAbs
11
66
PD-1 + OX40 agonist mAbs
1
4
PD-1 + Anti-Tim3 mAbs
23
142
Untreated
100
617
Tumor growth inhibition (T/C %) defined as the ratio of the median tumor
volumes of treated groups versus Vehicle group was calculated.
No R
No R
Ant-inf
Efficiency of combinations – EMT6
R
E
S
U
L
T
S
The minimum RTV in the untreated group defines the cutoff between responders (green) and non-responders (red).
No NR
(CD80) B7-1
(CD86) B7-2
Group
Cytokine concentration values have been normalized
by group using the following formula:
𝒗𝒗𝒗𝒗𝒗𝒗𝒗𝒗𝒗𝒗
𝒏𝒏𝒏𝒏𝒏𝒏𝒏𝒏𝒏𝒏𝒏𝒏𝒏𝒏𝒏𝒏𝒏𝒏𝒏𝒏 𝒗𝒗𝒗𝒗𝒗𝒗𝒗𝒗𝒗𝒗 = 𝒍𝒍𝒍𝒍𝒍𝒍
𝒎𝒎𝒎𝒎𝒎𝒎𝒎𝒎 𝒖𝒖𝒖𝒖𝒖𝒖𝒖𝒖𝒖𝒖𝒖𝒖𝒖𝒖𝒖𝒖𝒖𝒖 𝒈𝒈𝒈𝒈𝒈𝒈𝒈𝒈𝒈𝒈
T-cells
TCR
MHC class II
RTV Visualization
Cytokine quantification using multiplex assay
in the blood of mice the day of termination
(day 27).
8 cytokines are plotted (out of 25 measured).
Color of dots represent response : responders
(green) and non-responders (red).
EMT6
1 500
Untreated
Proportion of responders (green) vs non-responders (red).
Response was defined using relative tumor volume values
(RTV: tumor volume at end of experiment divided by
tumor volume at treatment start). Grey cells represent
animals with no available efficacy endpoint at the end of
the study.
American Association for Cancer Research (AACR) Annual Meeting / Washington, DC, USA. April 1-5, 2017
1 500
PD-1 mAb
1 500
CD137 mAb
1 000
1 000
1 000
1 000
500
500
500
500
0
0
0
0
0
4T1
1 500
Pro-inf
1 500
5
10
15
20
25
Time (Days)
30
35
1 500
Untreated
1 250
0
1 250
5
10
15
20
25
Time (Days)
30
35
0
1 500
PD-1 mAb
5
10
15
20
25
Time (Days)
30
1 500
CD137 mAb
1 250
1 000
1 000
750
750
750
750
500
500
500
500
250
250
250
250
5
10
15
20
25
Time (Days)
30
35
0
0
5
10
15
20
25
Time (Days)
30
35
0
0
5
10
15
20
25
Time (Days)
30
35
5
10
15
20
25
Time (Days)
30
CONCLUSIONS
35
PD-1 + CD137 mAbs
1 250
1 000
0
PD-1 + CD137 mAbs
0
35
1 000
0
Ad-Immu
Variation of cytokine profile with treatment response. For each
normalized cytokine value, the mean value of non-responders was
subtracted from the mean value of responders.
• Blue : R < NR
• Orange: R > NR
• White: N = R
Cytokine heatmaps. Normalized values were color-coded as follows:
• Blue: under-expressed values
• Orange : over-expressed values
• White : no difference
Tumor volume (mm3)
APCs
PD-1
Responder vs. non responder
Tumor volume (mm3)
I
N
T
R
O
D
U
C
T
I
O
N
PD-1 targeting antibodies (nivolumab, pembrolizumab) are now
approved in various tumor types either as second line treatment
(locally advanced or metastatic urothelial carcinoma, advanced
renal cell carcinoma, recurrent or metastatic head and neck
squamous cell carcinoma, classical Hodgkin lymphoma) or even as
first line therapy (metastatic melanoma, metastatic non-small cell
lung cancer).
Despite an increase in response rate as well as survival, there is still
issue with resistance to PD-1 targeting therapies. Selection
biomarkers as well as response biomarkers are still under intensive
research. In addition, combination strategies are needed to increase
response rate and overcome resistance.
EMT6 vs 4T1: CD137 mAb combined to PD-1 mAb
0
0
5
10
15
20
25
Time (Days)
30
35
 EMT6 model
• PD-1 mAb: global increase of the cytokine expression by the responders,
• Cytokine profiles after CD137 mAb treatment are different compared to all other tested mAbs,
• No obvious difference in cytokine profiles analyzed at the end of the study could be evidenced,
between responders and non-responders.
 EMT6 model compared to 4T1 model
• PD-1 mAb treatment induces profound changes in cytokine profiles,
• Cytokine profiles differ between a sensitive model EMT6, and a more resistant one 4T1.
 Summary
• Cytokines network is a complex system (that changes in a dynamic manner),
• Cytokine profile analysis combined with heatmap visualization are valuable tools,
 Additional data are needed to identify robust cytokine-based biomarkers
This work was partly supported by a grant from
and the French Government