Tumor Microenvironment Hijacking the Immune System [Read-Only]
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James H Finke Ph.D.
Department ImmunologyLerner ResearchCleveland Clinic
Tumor Microenvironment; Hijacking The Immune System
Disclosures
Research Support: Pfizer, GSK, Immatics and ChemDiv
Cellular Composition of the Tumor
• The tumor microenvironment is not only composed of malignant cells it contains:
• neovasculature, fibroblast and myeloid cells
• Immune cells critical for mounting an immune response to tumor.-T lymphocytes, CD4 and CD8 subsets- Dendritic cell subsets- NK/NKT cells -B cells/plasma cells
• Many of the tumor infiltrating immune cells are dysregulated, functionally impaired and contribute to tumor progression.
-T cells (anergic)-T-regulatory cells-Macrophages-Myeloid derived suppressor cells
Cellular Constituents of Immune Escape within the T umor MicroenvironmentKerkar SP, Restifo NP.Cancer Res. 2012 Jul 1;72(13):3125-30. Epub 2012 Jun 21. Review
• In metastatic RCC patients peripheral blood CD4+
and CD8+ T cells showed a skewing toward a TH-2
response against EphA2 and MAGE-6-derived
peptides except NED patients
Diminished Type 1 T cell Response in mRCC is Linked to Active Disease.
Tatsumi T et al. Cancer Res 2003;63:4481-4489
• CD8+ EphA2 specific T cells detected in
patients who are long term survivors
with residual disease and NED.
Features of some solid tumors that may mediate poor immune recognition or reduce immune destruction of tumor .
T cell Priming-Reduced recruitment of Dendritic subsets for antigen presentation-Inadequate expression of co-stimulatory molecules on tumor cells or infiltrating DC
Effector Phase-Inadequate recruitment of activated effector cells• chemokines
Presence of dominant immune inhibitor mechanisms th at suppress T cell effectorfunction• Suppressive cells (Treg, MDSC, Macrophages, neutrophils)• Inhibitory cytokines (IL-10, TGFβ)• Inhibitory receptors (CTLA4, PD1/PDL-1)
(n=22) (n=56) (n=40)
Tregs in Blood and Tumor of RCC patie nts
Finke J et al , unpublished and Clin Cancer Res 2008,
Treg Foxp3+ cells
A. B. T-regulatory cell facts
• Foxp3+ Treg cells suppress T effectors via different mechanisms .
• Difficult to selectively reduce Treg cell numbers.
• For adoptive T cell therapy the use of cytoablativestrategies to deplete Treq enhances clinical responses. (Dudley EM et al. J Clin Oncology 2008)
• Within different tumor types there is variable correlationbetween the degree of Treg infiltration and overall survival.
• In colon cancer Treg numbers have not correlated withreduced survival (Loddenkemper C. et al J Trans Med 2006).
Myeloid-derived Suppressor Cells
• Heterogeneous population of immunosuppressive myelo id cells• Normally present in very small amounts but systemic ally accumulate under pathologic conditions – tumor-bearing• Accumulation associated with:
VEGF, SCF, GM-CSF, G-CSF, S100A9, and M-CSF
• Murine MDSC: Granulocytic (CD11b+Gr1hi+),
Monocytic (CD11bGr1low)• Human MDSC multiple subsets:
Granulocytic (CD33lowHLADR-CD15+CD14-)Monocytic (CD33lowHLADR-CD15-CD14+)
Linage Negative (CD33lowHLADR-CD15-CD14-)
Plasticity of MDSCGranulocytic MDSC differentiate into CD31 Endothelial cells Monocytic MDSC differentiate into Tumor Associated MacrophageMonocytic MDSC differentiate into Granulocytic MDSC
Ko J et al Can Res 2010 and unpublished data
MDSC in Cancer Patients
MDSC subset levels in blood and tumor of RCC pts
Baseline levels of Monocytic and Granulocytic MDSC negatively correlate with overallsurvival
MDSC subset morphology .A.
B.
C.
G-MDSC M-MDSCLinage
Negative
G-MDSC Linage
Negative
M-MDSC
Walter S, et al Nat Med. 2012 Aug
Patient Granulocytic MDSC and Neutrophils: Suppress ive and Angiogenic Activities
% T
ce
ll
sup
pre
ssio
n
PatientG-MDSC Normal4:1 8:1 16:12:1 4:1 8:1 16:12:1 4:1 8:1 16:12:1
Neutrophil
T cell suppression
MDSC promote angiogenesis of RCC implanted in SCID mice
Ko J et al Manuscript in preparation
MDSC
ArginaseiNOS
InduceTreg
Macraphage M2 Cell IL-12
Th2Cell
IL-10
Tumor Progression
ArginineReductionCysteine Reduction
CD3ζ ζ ζ ζ CD4
T cell
CD3ζ ζ ζ ζ CD8
T cell
Cell Cycle Arrest
ROS NO (H2O2, peroxynitrate)
CD4T cell
CD8T cell
Apoptosis Nitrate TCR
MDSC –Mechanisms of Suppression
Granulocytic subsetROS, Arginase
Monocytic subset NO, Arginase
Neutrophils
• Elevated blood neutrophils and elevated neutrophil/lymphocytes ratio is associated with poor clinicaloutcome in several human cancers (RCC, melanoma colorectal, lung , ovarian etc) ((Donskov F. Seminar in Cancer Biology 2013)
• In non-metastatic localized clear cell RCC the presence of intratumoral CD66+ neutrophils was associated with short recurrence-free survival and overall survival (Jensen HK et al J Clin Oncology 2009)
• Neutrophils from cancer patients but not healthy donors can suppress T cell function and produce elevated levels of proangiogenic proteins (Schmielau J. and Finn O., Can Res 2001 , Rodriguez PC et al. Can Res 2009)
• The relationship between granulocytic -MDSC and patients neutrophils is being assessed (functional andgene array studies).
• Tumor microenvironment promotes pro-tumor neutrophils (TAN )Claudia A. Dumitru , et al Seminars in Cancer Biology Volume 23, Issue 3 2013 141 - 148
IL-1ββββTNFIL-6
IL-23IL-12
LPS IFNγγγγ/TNF
MHCII
M1 M2
IL-4, IL-10& IL-13
ImmuneComplexes+ IL-1/LPS
TGFββββIL-10
CCL22
Arg
MHCII
Functions
Stimuli Stimuli
Tumor Associated Macrophages
Type 1 Inflammation
Th1 ResponsesTumoricidal
Type II Inflammation
Th2 Responses
Tumor Promotion
Functions
Tissue Macrophages
PDL-1
Angiogenesis
Impair T Cell Activation
Macrophages
Gabrilovich et al Nat Rev Immunol. 2012 Mar 22;12(4):253-68
Multiple Subsets of TAM are Induced by the Tumor M icroenvironment
TAMs in RCC
RCC associated TAM can mediate immune suppression and tumor escape via the activation of the 15-Lipoxygenase-2 pathway.
Daukrin I. et al. Can Res ,2011
15-LOX
15-(S) HETEBioactive lipid
CCL2 MonocyteRecruitment
IL-10
T cell
FOXP3CTLA4Up-regulation
TAM
Tumor Hypoxia (Hif1-αααα)
Treg
CCL28
Angiogenesis
MDSC
Tumor Hypoxia (Hif1-αααα)
ArginaseNO Production
TAM
TAM
DCCD40CD80CD86MHC Class IICCR7
TNFαIL1βCCR5
SuppressiveFunction
PDL-1 (B7-H1) Expression by RCC and Its Suppression of T Cell Function
PDL-1
PDL-1
PDL-1
PDL-1 Expression in RCC-Poor Outcome
Thompson H et. al. Clin Can Res 13:1757-1761, 2007
Can be expressed on- Tumor- Macrophages-Dendritic Cells
Secreted form (sB7-H1)Immunosuppressive
Frigola X et al Clin Can Res 17:1915-1923,2011)
Gaurisankar Sa et al (Unpublished)
PD-L1
Bright Field
PDL-1 in tumor supernatant binds T cells which is blocked by anti-PDL-1 Ab
(Cancer J 2013;19: 353Y364)
Pre-Tx Cycle 1 Cycle 2 Cycle 4
G-MDSC ( ), CD3+ ( )
One Example of Modulating The Tumor Microenvironment : Sunitinib Treatment
Blood Tumor
Ko et a Clin Cancer Res. 15:2148-57, 2009 and unpublished data
5 10 15 20 25 300
500
1000
1500
2000
2500
3000UntreatedSUT OnlyVAC OnlySUT/VAC
1000
1500
MO
5 T
umor
Vol
ume
(mm
3 )
Days Post Tumor Inoculation
VAC
SUT
(0/30)
(0/30)
(0/30)
(12/50)
****** **
**
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5 10 15 20 25 300
500
1000
1500
2000
2500
3000UntreatedSUT OnlyVAC OnlySUT/VAC
1000
1500
MO
5 T
umor
Vol
ume
(mm
3 )
Days Post Tumor Inoculation
VAC
SUT
(0/30)
(0/30)
(0/30)
(12/50)
****** **
**
*
*
*
*
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*
Bose et al. Int. J. Cancer 2011; 129: 2158 - 2170.
Superior Anti-Tumor Efficacy of Vaccine + TKI Co-Therapy
MO5 (B16.OVA)
injected s.c .
(2 x 10 5)
+/- oral Sunitinib(0.1 mg/day, d10
+/- s.c . DC/OVA 257 -262
vaccines d10, d17
Monitor tumor size
TME analysis (d34)
Immune monitoring (d34)
MO5 (B16.OVA)
injected s.c .
(2 x 10 5)
+/- oral Sunitinib(0.1 mg/day, d10-16)
+/- s.c . DC1/OVA 257- 262
vaccines d10, d17
Monitor tumor size
TME analysis (d34)
Immune monitoring (d34)
Bose et al. Int. J. Cancer 2011; 129: 2158 - 2170.
Vaccine/TKI Co-Therapy Promotes the Inhibition of Regulatory Cells and the Activation/Recruitment of
Protective CD8+ T cells
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umor
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Abs
olut
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ell
Num
ber
in T
umor
(x 1
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Days Post-Inoculation
UntreatedSUT OnlyVAC OnlySUT/VAC
CD8-T CD4-TMDSC
(CD11b+Gr1+)
Treg (CD4+/ Foxp3 +)
DC (CD11c+/CD86+)
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umor
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umor
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Abs
olut
e C
ell
Num
ber
in T
umor
(x 1
06 )
Days Post-Inoculation
UntreatedSUT OnlyVAC OnlySUT/VAC
UntreatedSUT OnlyVAC OnlySUT/VAC
CD8-T CD4-TMDSC
(CD11b+Gr1+)
Treg (CD4+/ Foxp3 +)
DC (CD11c+/CD86+)
VEGF
HIF-1ααααHIF-2αααα
PDGFRββββ
Un
tre
ate
d
SU
T
VA
C
SU
T/V
AC
ββββ−−−−Actin
CXCL9
CXCL10
CXCL11
CXCR3IFN-γγγγ
Combination Therapy Results in a
Type-1 Biased Immune Profile in
the Tumor
Combination Treatment with Vaccine
and Sunitinib Improved T cell Response
Clinical Trials with Sunitinib Plus Vaccine therapy
Argos Therapeutics
DC loaded with autologous total tumor RNA + sunitinib
Immatics Biotechnology
Multipeptide vaccine + sunitinib
UPMC/Cleveland Clinic
DC pulsed with tumor blood vessel-associated antigens (TBVA) + sunitinib
Conclusions
• There is heterogeneity in patient response to immune-based Immunotherapy.
• A significant component of that heterogeneity comes from differences at the level ofthe tumor microenvironment.
• Key determining factors in the tumor (RCC) environment include recruitment of T effector cells, local production of chemokines and the presence of localimmunosuppressive mechanisms.
• Further Identifying the different mechanisms mediating immune suppression and angiogenesis by tumor stromal cells has already yielded new strategies forimproving cancer therapy (CTLA4 and PD1 antibodies) and is likely to
yield additional targets.
• The fact that different histological types of tumors all share in common the infiltrationof stromal cells should provide unique universal targets for therapy of solidtumors including RCC.
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