Preclinical imaging and therapy. Marion de Jong

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1 Preclinical imaging and therapy Marion de Jong

2 Content Introduction Preclinical Imaging Preclinical Therapy to raise awareness about problems related to translation of animal studies Radiopharmaceuticals (radiopeptides, targeted imaging and therapy) Radionuclides

3 Animals models and studies in translational reseach

4 Consensus on why and how? Quotes about animal studies: Animal research seldom applies to humans Dump animal research in favor of non-invasive methods Animal studies are still done only because people can make money from it

5 Translational research: crossing the valley of death Nature 453, , 2008 NIH: Lack of translation of basic research

6 Translational research In vivo molecular imaging in small animals is a bridge between in vitro data and translation to clinical application, but.

7 Be aware of differences in men vs mice Mice and humans are > 95% identical at the genomic level, their respective phenotypes are very different: Size, metabolism, hear beat rate, etc

8 Important differences Size Limitations for max volume injected or max volume of blood samples (about 200 µl/ mouse) Specific activity radiopharmaceuticals should be high, esp. in processes with low capacity, e.g. receptor binding. Heartbeat The heartbeat rate of a mouse : 600 beats/min Physiological processes occur faster Small animals can tolerate larger doses as they - clear most chemicals from their bodies much faster - given dose will lead to lower concentrations

9 Important differences Anaesthetics are known to alter animal physiology dramatically, cause changes in respiration, heart rate, blood pressure, and temperature.

10 J Nucl Med 2006; 47: , Fueger et al.

11 Study to determine optimal imaging conditions FDG IV iso IV awake IP iso IV awake 60min IV Fasted Yes Yes Yes No No Preheating 30 Yes Yes Yes No Yes no fasting Injection IV (iso) IV (awake) IP (iso) IV (awake) IV (iso) Method Cannula tail vein Needle tail vein Needle peritoneal Needle tail vein Cannula tail vein Awake after injection No No No 60 No Scantime MIP 180 ) White- Black: 0-15% max Bq/ml 90 scans shown

12 Performing animal imaging NanoSPECT/CT Equipped with heated rat- and mouse beds Connection for isoflurane anesthesia Isoflurane NanoSPECT/CT system Heating

13 Content Introduction Preclinical Imaging Preclinical Therapy Radiopharmaceuticals (radiopeptides, targeted imaging and therapy) Radionuclides

14 Targeted imaging and therapy Targets on tumor cells: extracellular and intracellular Extracellular Transporters Neurotransmitter receptors Hormone receptors Neuropeptide receptors Growth Factor receptors Antibody Epitopes Intracellular Metabolic pathways DNA/RNA Other organelles Courtesy Steve Mather

15 Targeted Imaging & Therapy: What are we trying to achieve? Selective imaging contrast and/or radiation dose: é as high as possible on the tumour ê as low as possible on normal tissues

16 How can we achieve this? Choose the best target Produce the best ligand Obtain the best tumor uptake/binding Treat the best patients: theranostics

17 Example: Regulatory peptides used/tested in the clinic as radiopeptides: Somatostatin neuroendocrine tumors CCK-B, gastrin MTC, SCLC GRP, Bombesin prostate, breast Neurotensin exocrine pancreatic tumors Substance P glioblastoma, astrocytoma RGD Newly formed blood vessels GLP-1 insulinomas, SCLC, carcinoid

18 Somatostatin analogues binding NE tumours Peptide Linker O HOOC C N D-Phe - Cys - Tyr N N H S D-Trp N N S Lys Thr- Cys -Thr HOOC COOH 90 Y 111 In 67/8 Ga 177 Lu Residualizing radionuclides

19 Diagnosis and Therapy 1. Diagnosis + 2. Therapy Theranostics

20 Diagnostic Radionuclides Gamma (γ) / positron (β + ) emitting radiometals for diagnostic radiopeptides Radiometal Half-life: t 1/2 (h) Decay mode E γ /E β+ (kev) Production 99m Tc 6.0 IT (100%) Mo/ 99m Tc generator 111 In 67.2 Auger, EC (100%) 172,247 cyclotron 67 Ga 78.1 Auger, EC (100%) 91,93,185,296,38 8 cyclotron 68 Ga 1.1 β + (90%) EC (10%) 820, Ge/ 68 Ga generator 64 Cu 12.9 β + (19%) β - (40%) EC (41%) 654 cyclotron, reactor 86 Y 14.7 β + (33%) EC (66%) 1043,1248,1603, 2019,2335 cyclotron EC: electron capture, IT: isomeric transition

21 Therapeutic Radionuclides Beta (β - ) emitting radiometals used for preparing therapeutic radiopeptides Radiometal Half-life, t 1/2 max β - (MeV) Decay mode γ (kev) Range (mm) Production 188 Re 17 h W/ 188 Re generator 186 Re 90.6 h reactor 90 Y 64.1 h 2.27 none Sr/ 90 Y generator 177 Lu 6.7 d 0.5 (79%) 208, reactor 67 Cu 61.9 h 0.58 (20%) 91,93,185, 395,484, accelerator

22 The Theranostic Approach : YouTube Movie Action BM0607, raise interest of the public Feel free to ask for the movie for eductional purposes

23 Content Introduction Preclinical Imaging Preclinical Therapy Radiopharmaceuticals (radiopeptides, targeted imaging and therapy) Radionuclides

24 Small animal imaging Scaling problems Volume: factor Length: factor 10-16

25 AMIE Facility

26 Preclinical PET versus SPECT Advantages of PET: higher sensitivity use of physiol. tracers better quantification less noise Advantages of SPECT tracers: have longer half-lives are cheaper more widely available higher resolution dual isotope studies PET SPECT Resolution 2 mm <1 mm Sensitivity pmol nmol Quantification ++ + Tracer half-lives sec s, hr s hr s, days Tracers Physiologic Modified Modified Physiologic Early kin. (min) Late kin. (days) +/- ++

27 MicroSPECT and micropet in Erasmus MC Inveon U-SPECT/CT Vector NanoSPECT/CT

28 Small animal PET/SPECT/CT imaging in oncological research Ø Pharmacokinetics and dosimetry (imaging, therapy) Ø Detection of metastases Ø Microdosimetry (kidneys, tumors) Ø Tumor response over time Ø Normal organ function over time

29 Courtesy: Dr. T. Maina-Nock Biodistribution study

30 Time activity curve Dynamic NanoSPECT/CT radiobombesin

31 Imaging of lung metastases, rat, Lu-octreotate + Rad001

32 Post therapy imaging: longitudinal Different tracers in one animal 278 MBq [ 177 Lu-DOTA]octreotate 5 days after therapy (injection radiopeptide) 50 MBq [ 111 In-DTPA]octreotide 4h after injection 28 days after therapy 50 MBq [ 99m Tc]DMSA 3h after injection 120 days after therapy

33 PC3 xenografts ( 111 In-bombesin) microdosimetry Sagittal Coronal Transversal Inhomogeneous tumour uptake of the radiopeptide: Receptor expression or perfusion?

34 Dynamic Contrast Enhanced-MRI Measure change in tissue relaxivity after injection of contrast agent Perfusion of contrast agent provides info on functional vessels

35 Maximum signal enhancement Overlay of maximum signal enhancement in tumor Contrast agent does not reach all areas of the tumor

36 DCE - SPECT SPECT MRI

37 Imaging 1) pharmacokinetics and dosimetry (imaging,therapy) 2) microdosimetry (kidneys, tumors) 3) effectiveness of e.g. kidney uptake reduction 4) tumor response over time 5) normal organ function over time

38 Peptide Receptor Imaging in small animals SPECT - Rel. low sensitivity: - specific act. should be high, radiochemical limitations! - high ligand amount: (partial) saturation of the receptor - high radioactivity dose PET - spatial resolution Longitudinal studies! Melis et al., J Nucl Med 51:973-7, 2010

39 177 Lu-production Conventional: 176 Lu +n/γ è 177 Lu NCA: 176 Yb +n/γ è 177 Yb (T½=1.9 h) è 177 Lu

40 177 Lu production Conventional: 176 Lu +n/γ è 177 Lu J in medium and low flux reactors L only 20% is converted è relatively low specific activity L 0.04 % of long-lived 177m Lu (T½=160 days) is produced NCA: 176 Yb +n/γ è 177 Yb (T½=1.9 h) è 177 Lu + β - J 100% yield è high specific activity achievable L high flux reactor L separation procedures

41 PRRT in CA20948 tumour-bearing rats Treatment None 278 MBq conventional 177 Lu-octreotate (11 µg) 278 MBq NCA 177 Lu-octreotate (2.1 µg) Monitoring 3x / week for 50 d p.t.: Tumour size (calliper) SPECT/CT day 1, 2, 4 and 8 p.t. Quantification of tumour uptake/volume Clearance rate and dosimetry

42 SPECT/CT day 1 p.t. Lu-octreotate Conv. 177 Lu-octreotate, 11 µg adrenals NCA 177 Lu-octreotate, 2 µg tumour tumour

43 Acknowledgements: Preclinical groups Nuclear Medicine/Radiology

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