Biomarkers from the Tumor Microenvironment: Are they Useful?
George Coukos, M.D, Ph.D.Director, Gynecologic Malignancy Research Programs
University of Pennsylvania
Biomarkers from the tumor microenvironment
• Tumor microenvironment events affect tumor outcome
• Tumor microenvironment events are identifiable through cellular or molecular markers
• Markers are mechanistically relevant• Modulation of tumor microenvironment affects
tumor outcome• Modulation of tumor microenvironment can be
traced through these markers
Tumor Microenvironment
Cancer CellsCancer Cells
Immune Cells
Stroma Cells
Endothelial Cells
The Reductionist View A Heterotypic Cell Biology
Do Leukocytes Play a Role in Ovarian Cancer ?
CD45 CD11c CD3
Cellular BiomarkersIntratumoral T cellsT regulatory cellsVascular leukocytes
CD3+
Intratumoral T cells
54.8%
38.7%
6.5%N=174
Zhang et al., NEJM 2003
Impact of TILs on Outcome in Ovarian CancerStage III/IV – All patients (n=174)
PFS Survival
+ TILs- TILs
N=174
Zhang et al., NEJM 2003
TILs correlate with tumor-reactive PBLs
Patient No. ELISPOT TILs
OV22 + +
OV23 -/+ +
OV24 -- --
OV26 -- --
OV27 + +
OV28 -- --
OV29 + +
OV31 + --
OV32 + +
OV34 -- --
P < 0.045
Intratumoral T cells exhibit oligoclonal expansion
A
B D
C
Vβ8
TILs Recognize TAAs in Ovarian Cancer
CD
8S
SC
CD
14-C
D4
Her2-369-APCEBV-PE TERT-PE Her2-689-PETax-PE
CD8FSC
.
Tumor Biopsyor Ascites
Disassociation
PE
APC-CY7
PE-Cy5
APC-Cy7
+Flow Cytometric
Analysis
Antigen Specific TILs
Cy5
PE
FreshlyIsolated
.
Bob Vonderheide
Proposed Biomarkers for Tumor Immune Surveillance
• Intratumoral CD8+ T cells
• IFN-γ, MIG, IP-10
• pLCK, pZap-70
• NKG2D ligands
CD4+CD25+ Treg
R3 live cells/CD3+
Ascites panel.019
1 10 100 1000 10000
FL2-H: CD25 PE
1
10
100
1000
10000
FL4-H: CD4 APC
R3 live cells/CD3+ Ascites panel.018
1 10 100 1000 10000 FL2-H: CD25 PE
1
10
100
1000
10000
FL4-H: CD8 APC
26.3
CD25
CD8 CD4
Numbe r of CD4+ cells added
0 1000 5000 10000 200000
20
40
60
80
100
120
CD4+C D25+
CD4+C D25-
B
% o
f M
axim
um
Res
pons
e
Numbe r of CD4+ cells added
0 1000 5000 10000 200000
20
40
60
80
100
120
CD4+C D25+
CD4+C D25-
B
% o
f M
axim
um
Res
pons
e
Numbe r of CD4+ cells added
0 1000 5000 10000 200000
20
40
60
80
100
120
CD4+C D25+
CD4+C D25-
CD4+C D25+
CD4+C D25-
B
% o
f M
axim
um
Res
pons
e
CD4+CD25+
CD4+CD25-
CTLA-4
CTLA-4
CD4
CD
25
CD4+CD25+
CD4+CD25-
CTLA-4
CTLA-4
CD4
CD
25
Woo et al., Cancer Res 2002; J Immunol 2003
Regulatory CD4+CD25+ T Cells in Ovarian Cancer
0
10
20
30
40
50
0 5 15 25 35 45 55 65 75 85 95
mRNA FOXP3 (M idpoint)
Pe
rce
nt
Curiel, Coukos et al., Nature Med 2004
Proposed Biomarkers for Tolerance
• CD4+CD25+ T cells
• foxP3
• GITR, CTLA-4
• Neuropilin-1
• IL-10, TGF-β• PD-1
CD11c+ cells with DC / endothelial phenotype and
vasculogenic competence
Vascular Leukocytes
Conejo-Garcia et al., Nature Med 2004
Ascites
Endothelial cells and vascular leukocytes coexist in human ovarian cancer
Conejo-Garcia et al., Blood 2004
Proposed Biomarkers for Angiogenesis
• CD45+CD11c+VE-cadherin+ cells
• VEGF-A
Activity of Biological Therapy in Ovarian Cancer
Hodi et al., PNAS 2003
0 1 2 3 4Years
0.2
0.4
0.6
0.8
1.0
PF
S (
%)
CP (n=68)
CP+IFN-γ (n=65)
P=0.031
Conejo-Garcia et al. Nat Med. 2004Windbichler GH, et al. Br J Cancer. 2000
Acknowledgements
Coukos Lab•Lin Zhang•Jose R. Conejo-Garcia•Fabian Benencia•Ron Buchanovitz•Alisha Mohamed•Cecilia Coureges•Heidi Gray•Ann Jenkins•David Holtz•Eugene Kang•Cristina Vezzani•Giorgia Regnani•Shun Liang•Karen Nishida
Dept. Biostatistics•Phyllis Gimotty
Abramson•Carl June
U. of Torino•Dionyssis Katsaros•Marco Massobrio