Transcript WP7_UZH_Rudin - Lungeninformationsdienst

WP7: Imaging cancer therapy
Katerina Dikaiou
Florian Stuker
Steffi Lehmann
Divya Vats
Sandra Buergi
Manoj Desai
Markus Rudin
FP7/FMTXCT
SNF
NCCR/SNF
FP7/LIVIMODE
NCCR/SNF
FP7/LIVIMODE
Institute for Biomedical Engineering UZH/ETH
Institute of Pharmacology and Toxicology UZH
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WP7: Imaging cancer therapy - Deliverables
7.1 Preparation of HIF transfected colon cancer cells (9)
7.2 a) Assessment and in-vivo imaging of HIF induction in sc implanted glioma cells stably
transfected with HIF reporter gene (15)
b) establishment of breast cancer model (24)
7.3 Characterization of the quantification accuracy of FMT-XCT vs. histological
correlates in assessing treatment of glioma & mammary cancrcinoma at primary site
metastatic sites in the lung. (32)
7.4 Characterization of the quantification accuracy of FMT-XCT vs. histological
correlates in assessing treatment of glioma & mamm. carcinoma mouse model (36)
7.5 Characterization of the quantification accuracy of FMT-XCT to resolve differential
treatment levels in glioma & mamm.carcinoma model in the presence or absence
anti-VEGF-R treatment as it relates to histological validation. (40)
7.6 Report on assessment of antiangiogenic therapy effects on HIF induction and
downstream readouts of HIF (induction of proangiogenic factors) with FMT-XCT (46).
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WP7: Imaging cancer therapy
1. FMT-MRI Prototype
2. Beta cell specific imaging: insulinoma
3. Hypoxia imaging: colon carcinoma
4. Outlook
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Multimodal imaging of hypoxia signaling & angiogenesis
parameter
biological information
Mod
vascular permeability
sites of angiogenesis
MRI
vumor blood volume
total vascularization
MRI
TBV and vessel size
vascular architecture
MRI
hypoxia
tumor oxygenation
PET
HIF1a and HIF activity
hypoxia signaling
OPT
VEGF
proangiogenic signaling
OPT
adhesion molecules
activated endothelium
OPT
proteases
ECM degradation
OPT
inflammatory cells
Immune response / ECM degradation
MRI
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process
angiogenic ‚endpoint‘:
vessel morphology
and physiology
angiogenic signaling
tumor-host tissue
interaction
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MRI readouts of angiogenesis
structure
vasculare leakage
tumor blood volume
high
low
2.00
3.00
1.50
-ln(S(t)/S0)
4.00
S(t)/S0-1
2.00
1.00
0.50
0.00
0.00
-1.00
-3.00
1.00
-0.50
0.00
3.00
6.00
9.00
12.00
15.00
18.00
-4.00
0.00
Time (min)
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4.00
8.00
12.00
Time (s)
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1. Multimodal imaging of in tumors (hybrid FMT-MRI)
F Stuker, K Dikaiou, C Baltes
FMT-MRI system
FMT-MRI detection setup
(adapted from Stuker et al. , Manuscript in prep)
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FMT-MRI Performance characterization
Spatial accuracy: in-plane and depth resolution
In-plane resolution
Quantitative analysis:
In-plane resolution
Depth accuracy
Depth accuracy
Dose linearity
(adapted from Stuker et al. , Manuscript in prep)
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Multimodal imaging of in tumors (hybrid FMT-MRI)
In vivo POC of simultaneous FMT-MRI measurement
Protease activity (Prosense®) in subcutaneously implanted C51 colon carcinoma
Limitations of current setup:
- FOV covered by detector array too small
- Dimension of detector (32x32) too small
- Reconstruction does not yet consider MRI
information
(adapted from Stuker et al. , Manuscript in prep)
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2. Target specific imaging of insulinoma: TMEM27
D Vats, K Dikaiou
Alexa 488
Alexa 488 + Dapi
Ins-1E-hTMEM
8/9mAb
Ins-1E
8/9mAb
Ins-1E-hTMEM
HA-mAb
10 m
(adapted from Vats et al. , Manuscript in prep)
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Target specific imaging of insulinoma: TMEM27
Fluoroscence intensity (a.u.)
fluorescence intensity (a.u.)
reconstructed
Fluoroscence intensity (a.u.)
2.0-210 - 2
2.0×10
TM EM /TM EM Alexa680-8/9-mAb
Ins-1E-hTMEM;
TM EM /TM EM
TM EM /Ha
Ins-1E-hTMEM;
TM EM /Ha Alexa680-anti-HA
Tumor/TM EM
Tumor/TM;EM
Ins-1E
Alexa680-8/9-mAb
1.510 - 2
1.5×10 -2
***
1.010 - 2
1.0×10 -2
***
5.010 - 3
5.0×10 -3
0
0
1
2
3
4
5
6
Days
Days after
injection
0 Vats et al. , Manuscript in prep)
(adapted from
0
1
2
3
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Days
4
5
6
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3. hypoxia inducible factor: HIF-1a regulation by pO2
S Lehmann, S Buergi, M Desai
O2
O2
FIH
HO-N
VHL
HO-N
VHL
HO-P
HO-P
a
a
PHD
a
b
a b
a b
Proteasomal degradation
HRE
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hypoxia inducible factor: HIF-1a regulation by pO2
regulation of HIF1a-mCherry fusion protein in cells
mouse embryonic fibroblasts
normoxic
induced (DMOG)
1) inhibition of degradation machinery stabilizes HIF1a reporter construct
2) stabilized product is found in cellular nucleus (transcription factor)
3) stabilzed product is transctiptionally active (expression of downstream genes)
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Multimodal imaging hypoxia signaling
Hypoxia: 18F-MISO PET
Hypoxia
hypoxia inducible factor (HIF1a)
HIF1a stability: luminescent reporter gene
HIF1a
transcription factor: HIF-1a/HIF1b
binding to
HRE-elements on DNA  Gene Expression
mCherry
fluc
viral
promoter
HIF activity: bioluminescent reporter gene
SV40
VEGF
vasodilation
i-NOS and HO-1
angiogenesis
EPO
erythropoiesis
fluc
HRE
VEGF
Angiogenesis: MRI vascular permeability
permeability
blood volume
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Multimodal imaging hypoxia signaling
multimodal hypoxia readouts
C51: HRE-luciferase
20
3.0
HIF1a-luc (group 1)
HIF1a-luc (group 2)
d6
hypoxia
pgl101-luc (group 1)
pgl101-luc (group 2)
d10
FMISO TMR (group 1)
2.0
FMISO TMR (group 2)
FMISO TMR
d8
HIF1a stability or
HIF transcriptional activity
15
10
1.0
5
HRE
HIF1a
d13
0
0.0
0
2
4
6
8
10
12
14
16
Days after inoculation
(adapted from Lehmann et al. , PNAS 2009)
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Multimodal imaging hypoxia signaling
day 7
8
9
11
14
H&E
pimonidazole
min
max
min
max
min
max
min
max
HIF1a
GLUT1
CD31
(adapted from Lehmann et al. , PNAS 2009)
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Multimodal imaging hypoxia signaling
Hypoxia
Non-hypoxic
HIF1a regulation
hypoxia inducible factor (HIF1a)
transcription factor: HIF-1a/HIF1b
binding to
HRE-elements on DNA  Gene Expression
VEGF
vasodilation
i-NOS and HO-1
angiogenesis
Feedback
EPO
erythropoiesis
permeability
blood volume
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Multimodal imaging hypoxia signaling
S Lehmann
(adapted from Lehmann et al. , Manuscript in prep)
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Multimodal imaging hypoxia signaling
50
45
40
35
30
25
20
15
10
5
0
(adapted from Lehmann et al. , Manuscript in prep)
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Outlook
1. Second generation FMT-MRI
- increased dimension of SPAD (FSt, KDi)
- improved reconstruction, irregular shaped object (KDi)
and accounting for tissue heterogeneity
- application: hypoxia signaling (FMT) and angiogenesis
(MRI)
2. Establishment of breast tumor model:
- sc and orthotopic (DVa)
- evaluation of novel protease probes (DVa, LIVIMODE)
3. Novel hypoxia assays using IFPs:
- fluorescence assays for HRE (SBu, MDe)
4. Orthotopic brain tumor model:
- hypoxia signaling in glioma models(SBu)
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Manuscripts
Manuscript published
S.Lehmann, D Stiehl, M Honer et al. Longitudinal and multimodal in vivo imaging of tumor hypoxia
and its downstream molecular events PNAS (2009)
Manuscript in preparation
S Lehmann, R Keist, M Rudin. GPI anchored avidin- a novel protein reporter for in vivo imaging
S Lehmann, D Stiehl, M Dominietto et al. Longitudinal imaging of the pharmacological inhibition of
PHDs in mouse tumor allografts
D Vats, H Wang, K Dikaiou et al. Multimodal imaging of pancreatic β-cells in vivo via TMEM27
antibody
F Stuker, C Baltes, K Dikaiou et al Hybrid small animal imaging system combining magnetic resonance
imaging with fluorescence tomography using single photon avalanche diode detectors
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