2012 02-27 eline vermeij

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2012 02-27 eline vermeij

  1. 1. In-vivo optical imaging in experimental arthritis modelsEline VermeijDepartment of RheumatologyPRIME lecture29-2-2012
  2. 2. Biophotonic imaging using the IVIS• Fluorescent & bioluminescent reporters• Detection in the range of 400-900 nm• Multiple animals (3 mice / 2 rats) at once• Minimally invasive• Quick: 1-10 minutes per measurement• Quantitative yet limited resolution
  3. 3. Rheumatoid Arthritis features
  4. 4. Rheumatoid Arthritis features Inducible promoters
  5. 5. Disease-inducible promoter reporters• In about 15-30% of the RA patients, the disease course is characterized by an intermittent pattern of exacerbation and remission• During disease course different genes are upregulated• Promoter of a gene consists of a unique combination of transcription factor binding sites• Molecular imaging of transcriptional regulation of these genes during disease can be done by using expression vectors
  6. 6. Disease-inducible promoter reporters• Regulation of the expression of the genes in-vivo is largely unknown and very complex• With these promoter reporters we can mimic gene expression• By using quantitative imaging we can visualize and calculate the upregulation of genes Geurts et al., 2009.
  7. 7. Disease-inducible promoter reporters• Selected 10 different disease-inducible promoter reporters  in-vivo testing• Synovial lining targeted via viral transduction  intra-articular injection• Streptococcal Cell Wall arthritis (SCW)
  8. 8. In-vivo luciferase measurementsSAA3
  9. 9. In-vivo luciferase measurements Histological scoringSAA3
  10. 10. In-vivo luciferase measurements Histological scoringSAA3S100A8
  11. 11. In-vivo luciferase measurements Histological scoringSAA3S100A8
  12. 12. Disease-inducible promoters are a valuable imaging tool to monitor arthritis activity• Imaging of the activation of cells in the synovial lining / gene expression • Follow up during disease progression • Treatment respons
  13. 13. Rheumatoid Arthritis features Activatable and targeting NIR-probes
  14. 14. Fluorescence imaging of activatable and targeting probes• ProSense: Activated by cathepsin B, S, L, K  inflammation• MMPSense (Metalloproteinases): Activated by MMP 2, 9, 13  bone and cartilage destruction• OsteoSense: Targets hydroxyapatite (bone mineral)  active bone remodeling
  15. 15. Fluorescence imaging of enzyme activity and treatment response in a mouse model of RA• Collagen-induced arthritis  chronic arthritis model• IL-1 important pro-inflammatory cytokine • Pre-treatment with anti-IL-1 should diminish disease and enzyme activity and therefore fluorescent signal intensity
  16. 16. Fluorescence imaging of enzyme activity and treatment response in a mouse model of RA• Collagen-induced arthritis  chronic arthritis model• IL-1 important pro-inflammatory cytokine • Pre-treatment with anti-IL-1 should diminish disease and enzyme activity and therefore fluorescent signal intensity Fluorescence imaging is a valuable tool to monitor the protective effect of anti-IL-1 treatment
  17. 17. MMP activity in mouse model of OA• Osteoarthritis (OA) is characterized by cartilage damage and bone spurs• Less inflammation compared to rheumatoid arthritis • MMP activity (cartilage and bone damage) expected, but no cathepsin activity (inflammation)
  18. 18. MMP activity in mouse model of OA• DMM model • Destabilization of the medial meniscus • Cartilage damage • MMP activity in OA process?• MMPSense and ProSense imaging at week 8
  19. 19. MMP activity in mouse model of OA• DMM model • Destabilization of the medial meniscus • Cartilage damage • MMP activity in OA process?• MMPSense and ProSense imaging at DMM week 8 MMPSense week 8 DMM/Control = 1.23 MMPSense ProSense
  20. 20. MMP activity in mouse model of OA• DMM model • Destabilization of the medial meniscus • Cartilage damage • MMP activity in OA process?• MMPSense and ProSense imaging at DMM week 8 MMPSense week 8 DMM/Control = 1.23 MMPSense ProSense MMP activity but no cathepsin activity during OA process in DMM model
  21. 21. ProSense and MMPSense in SCID model• SCID model is a humanized mouse model for rheumatoid arthritis• Transplantation of synovium from RA patients to immunodeficient SCID mice• This model can be used to screen the effect of different therapeutics on human material• Protocol: • D0 = transplantation of biopsies subcutaneously on the back (ø 6mm) • D 0-7 = engraftment period • D 7-14 = treatment period • D14 = imaging and sacrifice: collection of blood and synovial grafts hIL-6
  22. 22. ProSense and MMPSense in SCID model• Imaging with ProSense (inflammation) and MMPSense (MMP activity) at 14 days after transplantation• Correlation ProSense with pro-inflammatory cytokine IL-8 in serum (r=0.95) ProSense
  23. 23. ProSense and MMPSense in SCID model• Imaging with ProSense (inflammation) and MMPSense (MMP activity) at 14 days after transplantation• Correlation ProSense with pro-inflammatory cytokine IL-8 in serum (r=0.95) ProSense MMPSense
  24. 24. ProSense and MMPSense in SCID model• Imaging with ProSense (inflammation) and MMPSense (MMP activity) at 14 days after transplantation• Correlation ProSense with pro-inflammatory cytokine IL-8 in serum (r=0.95) ProSense MMPSense NIR-fluorescent probes are useful tools to measure enzyme activity in real-time and quantitatively
  25. 25. Acknowledgements Department of Rheumatology Radboud University Nijmegen Medical Centre • Onno Arntz • Miranda Bennink • Marije Koenders • Wim van den Berg • Fons van de Loo

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