WP7_UZH_Rudin - Lungeninformationsdienst
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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 FMTXCT Meeting / IESL-FORTH Heraklion / 26032010 1 / 25 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). FMTXCT Meeting / IESL-FORTH Heraklion / 26032010 2 / 25 WP7: Imaging cancer therapy 1. FMT-MRI Prototype 2. Beta cell specific imaging: insulinoma 3. Hypoxia imaging: colon carcinoma 4. Outlook FMTXCT Meeting / IESL-FORTH Heraklion / 26032010 3 / 25 FMTXCT Meeting / IESL-FORTH Heraklion / 26032010 4 / 25 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 FMTXCT Meeting / IESL-FORTH Heraklion / 26032010 process angiogenic ‚endpoint‘: vessel morphology and physiology angiogenic signaling tumor-host tissue interaction 5 / 25 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) FMTXCT Meeting / IESL-FORTH Heraklion / 26032010 4.00 8.00 12.00 Time (s) 6 / 25 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) FMTXCT Meeting / IESL-FORTH Heraklion / 26032010 7 / 25 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) FMTXCT Meeting / IESL-FORTH Heraklion / 26032010 8 / 25 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) FMTXCT Meeting / IESL-FORTH Heraklion / 26032010 9 / 25 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) FMTXCT Meeting / IESL-FORTH Heraklion / 26032010 10 / 25 Target specific imaging of insulinoma: TMEM27 Fluoroscence intensity (a.u.) fluorescence intensity (a.u.) reconstructed Fluoroscence intensity (a.u.) 2.0-210 - 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 1.510 - 2 1.5×10 -2 *** 1.010 - 2 1.0×10 -2 *** 5.010 - 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 FMTXCT Meeting / IESL-FORTH Heraklion / 26032010 Days 4 5 6 11 / 25 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 FMTXCT Meeting / IESL-FORTH Heraklion / 26032010 12 / 25 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) FMTXCT Meeting / IESL-FORTH Heraklion / 26032010 13 / 25 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 FMTXCT Meeting / IESL-FORTH Heraklion / 26032010 14 / 25 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) FMTXCT Meeting / IESL-FORTH Heraklion / 26032010 15 / 25 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) FMTXCT Meeting / IESL-FORTH Heraklion / 26032010 16 / 25 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 FMTXCT Meeting / IESL-FORTH Heraklion / 26032010 17 / 25 Multimodal imaging hypoxia signaling S Lehmann (adapted from Lehmann et al. , Manuscript in prep) FMTXCT Meeting / IESL-FORTH Heraklion / 26032010 18 / 25 Multimodal imaging hypoxia signaling 50 45 40 35 30 25 20 15 10 5 0 (adapted from Lehmann et al. , Manuscript in prep) FMTXCT Meeting / IESL-FORTH Heraklion / 26032010 19 / 25 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) FMTXCT Meeting / IESL-FORTH Heraklion / 26032010 20 / 25 FMTXCT Meeting / IESL-FORTH Heraklion / 26032010 21 / 25 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 FMTXCT Meeting / IESL-FORTH Heraklion / 26032010 22 / 25
