WP7: Imaging cancer therapy

1. WP7: Imaging cancer therapy Katerina Dikaiou FP7/FMTXCT Florian Stuker SNF Steffi Lehmann NCCR/SNF DivyaVats FP7/LIVIMODE Sandra Buergi NCCR/SNF ManojDesaiFP7/LIVIMODE Markus Rudin Institute for Biomedical Engineering UZH/ETH Institute ofPharmacologyandToxicology UZH

2. 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.carcinomamodel 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).

3. WP7: Imaging cancer therapy FMT-MRI Prototype Beta cellspecificimaging: insulinoma Hypoxiaimaging: coloncarcinoma Outlook

5. Multimodal imagingofhypoxiasignaling & angiogenesis

6. MRI readoutsofangiogenesis structure vasculareleakage tumorbloodvolume high low

7. 1. Multimodal imagingof in tumors (hybrid FMT-MRI) F Stuker, K Dikaiou, C Baltes FMT-MRI detectionsetup FMT-MRI system (adaptedfrom Stuker et al. , Manuscript in prep)

8. FMT-MRI Performance characterization Spatialaccuracy: in-plane anddepthresolution In-plane resolution Depthaccuracy Quantitative analysis: In-plane resolution Depthaccuracy Dose linearity (adaptedfrom Stuker et al. , Manuscript in prep)

9. Multimodal imagingof in tumors (hybrid FMT-MRI) In vivo POC ofsimultaneous FMT-MRI measurement Proteaseactivity (Prosense®) in subcutaneouslyimplanted C51 coloncarcinoma Limitationsofcurrentsetup: - FOV coveredbydetectorarraytoosmall - Dimension ofdetector (32x32) toosmall - Reconstructiondoes not yetconsider MRI information (adaptedfrom Stuker et al. , Manuscript in prep)

10. 2. Target specificimagingofinsulinoma: TMEM27 D Vats, K Dikaiou Alexa 488 Alexa488 + Dapi Ins-1E-hTMEM 8/9mAb Ins-1E 8/9mAb Ins-1E-hTMEM HA-mAb (adaptedfromVats et al. , Manuscript in prep) 10 m

11. Target specificimagingofinsulinoma: TMEM27 Ins-1E-hTMEM; Alexa680-8/9-mAb Ins-1E-hTMEM; Alexa680-anti-HA Ins-1E ; Alexa680-8/9-mAb (adaptedfromVats et al. , Manuscript in prep)

12. 3. hypoxia inducible factor: HIF-1aregulation by pO2 S Lehmann, S Buergi, M Desai O2 O2 PHD a b FIH HO-N HO-N b a a VHL HO-P HO-P a VHL a b HRE Proteasomaldegradation

13. regulationof HIF1a-mCherry fusionprotein in cells hypoxia inducible factor: HIF-1aregulation by pO2 mouse embryonic fibroblasts normoxic induced (DMOG) 1) inhibitionofdegradationmachinerystabilizesHIF1areporterconstruct 2) stabilizedproductisfound in cellularnucleus (transcriptionfactor) 3) stabilzedproductistransctiptionallyactive (expressionofdownstream genes)

14. SV40 fluc Multimodal imaginghypoxiasignaling Hypoxia: 18F-MISO PET Hypoxia hypoxiainduciblefactor (HIF1a) transcriptionfactor: HIF-1a/HIF1b bindingtoHRE-elements on DNA Gene Expression HIF1astability: luminescentreportergene mCherry HIF1a fluc viral promoter HIF activity: bioluminescentreportergene VEGF i-NOS and HO-1 EPO HRE VEGF Angiogenesis: MRI vascularpermeability angiogenesis vasodilation erythropoiesis permeability bloodvolume

15. hypoxia HIF1a Multimodal imaginghypoxiasignaling multimodal hypoxia readouts C51: HRE-luciferase d6 d8 d10 HRE d13 (adaptedfrom Lehmann et al. , PNAS 2009)

16. max max max max min min min min Multimodal imaginghypoxiasignaling day 7 8 9 11 14 H & E pimonidazole HIF1a GLUT1 CD31 (adaptedfrom Lehmann et al. , PNAS 2009)

17. Multimodal imaginghypoxiasignaling Hypoxia hypoxiainduciblefactor (HIF1a) transcriptionfactor: HIF-1a/HIF1b bindingtoHRE-elements on DNA Gene Expression Non-hypoxic HIF1a regulation Feedback VEGF i-NOS and HO-1 EPO angiogenesis vasodilation erythropoiesis permeability bloodvolume

18. Multimodal imaginghypoxiasignaling S Lehmann (adaptedfrom Lehmann et al. , Manuscript in prep)

19. Multimodal imaginghypoxiasignaling (adaptedfrom Lehmann et al. , Manuscript in prep)

20. Outlook Second generation FMT-MRI - increaseddimensionof SPAD (FSt, KDi) - improvedreconstruction, irregularshapedobject (KDi) andaccountingfortissueheterogeneity - application: hypoxiasignaling (FMT) andangiogenesis (MRI) Establishment ofbreasttumor model: - scandorthotopic (DVa) - evaluationofnovelproteaseprobes (DVa, LIVIMODE) Novelhypoxiaassaysusing IFPs: - fluorescenceassaysfor HRE (SBu, MDe) 4. Orthotopicbraintumor model: - hypoxiasignaling in gliomamodels(SBu)

22. Manuscripts Manuscriptpublished 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 tumorallografts 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