wp7: imaging cancer therapy

FMTXCT Meeting / IESL-FORTH Heraklion / 26032010 1 / 25

WP7: Imaging cancer therapy

Katerina Dikaiou FP7/FMTXCTFlorian Stuker SNFSteffi Lehmann NCCR/SNFDivya Vats FP7/LIVIMODESandra Buergi NCCR/SNFManoj Desai FP7/LIVIMODEMarkus Rudin

Institute for Biomedical Engineering UZH/ETHInstitute of Pharmacology and Toxicology UZH


WP7: Imaging cancer therapy - Deliverables7.1 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).


WP7: Imaging cancer therapy

1. FMT-MRI Prototype

2. Beta cell specific imaging: insulinoma

3. Hypoxia imaging: colon carcinoma

4. Outlook


Multimodal imaging of hypoxia signaling & angiogenesis

parameter biological information Mod process

vascular permeability sites of angiogenesis MRI angiogenic ‚endpoint‘: vessel morphology and physiology

vumor blood volume total vascularization MRI

TBV and vessel size vascular architecture MRI

hypoxia tumor oxygenation PETangiogenic signalingHIF1a and HIF activity hypoxia signaling OPT

VEGF proangiogenic signaling OPT

adhesion molecules activated endothelium OPTtumor-host tissue interactionproteases ECM degradation OPT

inflammatory cells Immune response / ECM degradation MRI


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structure vasculare leakage tumor blood volume

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MRI readouts of angiogenesis


1. Multimodal imaging of in tumors (hybrid FMT-MRI)

FMT-MRI system

FMT-MRI detection setup

F Stuker, K Dikaiou, C Baltes

(adapted from Stuker et al. , Manuscript in prep)


FMT-MRI Performance characterizationSpatial accuracy: in-plane and depth resolution

In-plane resolution Depth accuracy Dose linearityQuantitative analysis:

Depth accuracyIn-plane resolution



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



Ins-1E-hTMEMHA-mAb

Ins-1E-hTMEM8/9mAb

Ins-1E8/9mAb

Alexa 488 Alexa 488 + Dapi

10 m

2. Target specific imaging of insulinoma: TMEM27D Vats, K Dikaiou

(adapted from Vats et al. , Manuscript in prep)


0 1 2 3 4 5 60

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TMEM/TMEMTMEM/HaTumor/TMEM

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Days after injection

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TMEM/TMEMTMEM/HaTumor/TMEM

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Ins-1E-hTMEM; Alexa680-8/9-mAb

Ins-1E-hTMEM; Alexa680-anti-HA

Ins-1E ; Alexa680-8/9-mAb

Target specific imaging of insulinoma: TMEM27

(adapted from Vats et al. , Manuscript in prep)


3. hypoxia inducible factor: HIF-1a regulation by pO2

HO-N

HO-P

HO-N

HO-P

a

a

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HRE

PHDFIH

VHL

VHL

Proteasomal degradation

O2

O2

S Lehmann, S Buergi, M Desai


regulation of HIF1a-mCherry fusion protein in cells

induced (DMOG)normoxic

mouse embryonic fibroblasts

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)

hypoxia inducible factor: HIF-1a regulation by pO2


viral promoter

HIF1a mCherry

permeabilityblood volume

erythropoiesisangiogenesisvasodilation

i-NOS and HO-1VEGF EPO

Hypoxia

hypoxia inducible factor (HIF1a)

transcription factor: HIF-1a/HIF1b

binding to HRE-elements on DNA Gene Expression

SV40fluc

HRE VEGF

Hypoxia: 18F-MISO PET

fluc

HIF1a stability: luminescent reporter gene

HIF activity: bioluminescent reporter gene

Angiogenesis: MRI vascular permeability

Multimodal imaging hypoxia signaling


hypoxia

HIF1aHRE

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Days after inoculation

HIF

1a s

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FMIS

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HIF1a-luc (group 1)

HIF1a-luc (group 2)

pgl101-luc (group 1)

pgl101-luc (group 2)

FMISO TMR (group 1)

FMISO TMR (group 2)

multimodal hypoxia readouts

d6

d10

d8

d13

C51: HRE-luciferase


(adapted from Lehmann et al. , PNAS 2009)


maxmin

maxmin

maxmin

maxmin

H & E

pimonidazole

HIF1a

GLUT1

CD31

day 7 8 9 11 14

(adapted from Lehmann et al. , PNAS 2009)



permeabilityblood volume

erythropoiesisangiogenesisvasodilation

i-NOS and HO-1VEGF EPO

Hypoxia

hypoxia inducible factor (HIF1a)

transcription factor: HIF-1a/HIF1b

binding to HRE-elements on DNA Gene Expression


Feedback

Non-hypoxic HIF1a regulation


Multimodal imaging hypoxia signalingS Lehmann

(adapted from Lehmann et al. , Manuscript in prep)


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35

40


(adapted from Lehmann et al. , Manuscript in prep)


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)

Outlook


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

Manuscripts

wp7: imaging cancer therapy

Documents

prepfmtxct meeting iesl

quantification accuracy

multimodal imaging of

insulinoma hypoxia imaging

carcinoma model

hif reporter gene

histological correlates

carcinoma mouse model