2012 02-27 eline vermeij
DESCRIPTION
http://www.umcn.nl/Research/Departments/cdl/PRIME/Pages/PRIMELecture.aspxTRANSCRIPT
In-vivo optical imaging in experimental
arthritis models
Eline Vermeij
Department of Rheumatology
PRIME lecture
29-2-2012
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
Rheumatoid Arthritis features
Rheumatoid Arthritis features
Inducible promoters
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
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.
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)
In-vivo luciferase measurements
SAA3
In-vivo luciferase measurements Histological scoring
SAA3
In-vivo luciferase measurements Histological scoring
S100A8
SAA3
In-vivo luciferase measurements Histological scoring
S100A8
SAA3
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
Rheumatoid Arthritis features
Activatable
and targeting
NIR-probes
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
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 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
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)
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
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
DMM week 8
MMPSense
DMM/Control = 1.23
MMPSense ProSense
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
DMM week 8
MMPSense
DMM/Control = 1.23
MMP activity but no cathepsin activity during OA process in DMM model
MMPSense ProSense
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:
• D 0 = 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
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
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)
MMPSense ProSense
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)
NIR-fluorescent probes are useful tools to measure enzyme activity in
real-time and quantitatively
MMPSense ProSense
Acknowledgements
Department of Rheumatology
Radboud University Nijmegen
Medical Centre
• Onno Arntz
• Miranda Bennink
• Marije Koenders
• Wim van den Berg
• Fons van de Loo