AXL Inhibition Improves Immunotherapy by …...Agata Rybicka3, Muntequa I. Siraji1,2, Sushil...
Transcript of AXL Inhibition Improves Immunotherapy by …...Agata Rybicka3, Muntequa I. Siraji1,2, Sushil...
References: 1Gubin et al., 2018. 2Chen and Mellman, 2017. 3Lemke and Rothlin, 2008. 4Ye et al., 2010. 5Chiu et al., 2015. 6Davidsen K.T et al., 2017. 7Terry et al., 2017.
ICB + bemcentinib targets tumor-immune crosstalk AXL is highly expressed in suppressive myeloid cells AXL+ M2φs are reduced by bemcentinib treatmentAXL is expressed in distinct DC populations with highly suppressive profiles
Conclusions
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NK cells
NK-S1
NK-S2NK-S3
NK-S5
CD11b-
CD11b+
NK-S4 Ly6G
+
MHC
II
+
-
iNOS+pDCs
DC-S9
DC-S8
CD4+
CD4-
DC
-S5
CD4+ DC-S4
CD206+ DC-S7
cDC2
DC-S1
CD80+
DC-S6
DC-S3
DC-S2
CD24+CD8+cDC1
Neutrophils
Immature cDC2
PD
L1high
DC-T3DC-T4
Ly6C+(pDCs)
CD4+
CD4-
DC-T1
Neutrophils
CD103+cDC1
CD80 low
DC-T8
DC-T9
CD80 int
CD11b+MHCII-CD24+
DC-T2
cDC2
DC-T5
DC-T6
DC-T7
Arg
-1+
Axl
+
NK cells
NK-T2
NK-T3
NK-T
1
CD11bCD
11b-
+
-
Ly6C
+
-
Spleen Tumor
Force-directed layout of X-Shift clusters visualizes rare cell populations in spleens and tumors.
Lorem ipsum
AXL is Expressed in Rare Dendritic Cell Populations
T1Sp
leen
Axl MerTK
T2Sp
leen
Axl MerTK
Tumo
rT1
T2Tu
mor
Uniform Manifold Approximation and Projection (UMAP ) of single immune cells showing AXL and MerTK expression at timepoint 1 and timepoint 2 in spleens and tumors.
Tumors mobilize AXL in Suppressive Myeloid Cells
CD4 T cells
CD8 T cells
gMDSC
NK cells
B cells
Macrophages/monocytes
Macrophages/monocytes
T1
T2
Spleen Tumor
BALB/C mice bearing established 4T1 mammary tumors were treated as indicated, n=5 per treatment per timepoint. Spleens and tumors were analyzed by CyTOF using a 34- marker myeloid focused panel. Graphs show single immune cells mapped with UMAP (left; colored by marker expression of CD11b, right;colored by density showing changes over time in immune populations in untreated animals)
Day 0 Day 12 Day14 Day 16 Day 18 Day 22
αCTLA4/αPD1
Bemcentinib
Rand
omiza
tion
T1T2
ET2ET1
High Dimensional Single Cell Immune Phenotyping
CSF1
mRN
A fol
d cha
nge
relat
ive to
ctr
0
10
20
30
CSF1
αCTLA4/ αPD 1 αCTLA4/ αPD1/Bem
Non- responder s Responder s
P = 0.0416 Fold change of CSF1 gene expression in 4T1 tumors unresponsive to ICB (n=4) or ICB and bemcentinb (n=5) and responsive tumors treated with ICB/bemcentinib (n=2), compared to vehicle
AXL Inhibition Blocks Tumor-Myeloid Crosstalk
0 50 100 1500
50
100
Days of treatment
Perce
ntsu
rviva
l
Vehicle n= 1 1Bemcentinib n=1 2
αCTLA4 /αPD1 n=17αCTLA4 /αPD1/Bemcentinib n=2 6
p = 0.0189
Durable tumor clearance in 25.6% of animalswith established orthotopic 4T1 mammary tumors treated with bemcentinib + anti-CTLA-4 + anti-PD-1 vs 5.6% of animals treated with anti-CTLA-4 + anti-PD-1
AXL Inhibition Potentiates the Effect on Immune Checkpoint Inhibitors
Tumor
Single immune cells clustered with PhenoGraph and analyzed with Marker Enrichment Modeling (MEM). Heat map of MEM-scores with changes in individual PhenoGraph clusters over time and between treatments.
619141334
122320252
1617101
159
2411217
1858
22
CD
206
CD
11b
CD
3
CD
4
Ly6C
MH
CII
CD
80Ly
6G-C
CD
24
CD
40iN
OS
CD
11c
CD
19
CD
25A
xl
Mer
TK
CD
49b
CD
69
CD
86
CD
45P
D1
CTL
A4
CD
83
CD
8a
PD
L1
CD
335
CD
114
Arg
-1
CD
115
CD
64
F4/8
0
CD
103
CD
116
CD
206
CD
11b
CD
3
CD
4
Ly6C
MH
CII
CD
80Ly
6G-C
CD
24
CD
40iN
OS
CD
11c
CD
19
CD
25 Axl
Mer
TK
CD
49b
CD
69
CD
86
CD
45P
D1
CTL
A4
CD
83
CD
8a
PD
L1
CD
335
CD
114
Arg
-1
CD
115
CD
64
F4/8
0
CD
103
CD
116
T1 T2
Ctr
Bem
ICB
ICB
+ B
em
ICB
+ B
emCtr
Bem
ICB
gMDSC
NK cells
DCs
Mφ
mMDSC
CD4 T cells
Axl+MerTK+Mφ/M2φ
gMDSC
pDCs
Lin-
B cells
B cells
Axl+MerTK+Mo/Mφ
CD8 T cellsCD4-CD8-T cells
CD
116
CD
80C
D11
cF4
/80
CD
206
CD
64M
HC
IIC
D11
bC
D24
Ly6G
-CLy
6C Axl
Mer
TKiN
OS
Arg
-1C
D11
5C
D69
PD
L1C
D10
3C
D86
CD
3C
TLA
4C
D49
bC
D40
CD
45P
D1
CD
83C
D33
5C
D25
CD
114
CD
19C
D4
CD
8
CD
116
CD
80C
D11
cF4
/80
CD
206
CD
64M
HC
IIC
D11
bC
D24
Ly6G
-CLy
6CA
xlM
erTK
iNO
SA
rg-1
CD
115
CD
69P
DL1
CD
103
CD
86C
D3
CTL
A4
CD
49b
CD
40C
D45
PD
1C
D83
CD
335
CD
25C
D11
4C
D19
CD
4C
D8
1819
136
20
1115
16251410
2422
85
12
34
97
1723
Axl+ M2φint. Mφ
iNOS+Arg-1+MφMerTK+M2φ
M2φ
Ly6C high Mo/MφCD49b+Axl+MDSC
CD24+ Mo/Mφ
gMDSC
pDCLy6C+ CD8 T cells
DCsMo/Mφ
Lin-
CD4-CD8-T cells
CD8 T cells
CD4 T cells
NK cellsLin-
T1 T2
Ctr
Bem
ICB
ICB
+ B
em
ICB
+ B
emCtr
Bem
ICB
Spleen
AXL Inhibition Reshapes the Immune LandscapeBackground
Tumor mobilization of suppressive myeloid cells remains a primary obstacle to immune checkpoint blockade (ICB) efficacy(1).
Tumor microenvironments enriched in suppressive myeloid cells are often T cell infiltrated(2) providing a rationale for ICB combination strategies with myeloid targeting agents.
The receptor tyrosine kinase AXL is a key tolerogenic regulator of the innate immune response(3).
Tumor associated macrophages (TAM) express AXL(4) and mediates M2-macrophage polarization(5).
In cancer AXL is associated with epithelial-to-mesenchymal (EMT) -related phenotypic plasticity and therapy resistance (6).
EMT is increasingly recognized as a major contributor to tumor immune evasion(7).
The small molecule AXL inhibitor bemcentinib targets tumor cell intrinsic phenotypic plasticity and myeloid immune suppression.
Bemcentinib in combination with ICB is currently in phase II trials for NSCLC (NCT03184571), melanoma (NCT02872259) and mesothelioma (NCT0365483).
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AXL Inhibition Improves Immunotherapy by Targeting Local and Systemic Tumor Myeloid Cross-Talk Kjersti T.Davidsen1,2, Katarzyna Wnuk-Lipinska3, Sturla M. Grøndal1,2, Noelly Madeleine1,2, Stacey A. D’mello Peters1,2, Magnus Blø3, Linn Hodneland3, Lavina Ahmed3, Agata Rybicka3, Muntequa I. Siraji1,2, Sushil Dhaka1,2, Oddbjørn Straume2,4, Michael A. Curran5, Rolf A.Brekken6, Gro Gausdal3, James B. Lorens1,2
1 Department of Biomedicine, 2 Centre for Cancer Biomarkers, University of Bergen, 3 BerGenBio ASA, 4 Department of Oncology, Haukeland University Hospital, 5 Department of Immunology, The University of Texas MD Anderson Cancer Center, 6 Department of Surgery and Hamon Center for Therapeutic Oncology Research, UT Southwestern Medical Center