animal imaging: cf gene therapy departments of radiology, biomedical engineering, and general...
TRANSCRIPT
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Animal Imaging: CF Gene Therapy
Departments of Radiology, Biomedical Engineering, and General medical
Science
Case Western Reserve University
Cleveland, OH 44106
Zhenghong Lee, Ph.D.
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Acknowledgement
CF Center (CWRU)
AG Ziady, Ph.D.PB Davis, M.D., Ph.D.
JLab (DOE)
S. Majewski, Ph.D.D. Weisenberger, Ph.D.
Radiology/BME (CWRU)
GR WojtkiewiczJJ DerakshanPF MartoneM Kotlarchyk
Univ. of Virginia
Dongfeng Pan, Ph.D.UCLA
H. Herschman, Ph.D.Q. Liang, Ph.D.
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Molecular/Small Animal Imaging
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Planar Multimodal Imaging System
Small animal scanner,developed by JLab at
CWRU/UHC, combining x-ray and scintigraphy
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CF Gene Therapy Measurement
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Molecular conjugate-DNA complex construction
+ + + +
+++
Plasmid DNA
Poly-L-lysine (~256 K)/LigandConjugate
DNA-Carrier Complex
aCl
Ethidium azide FITC
25 nm
C105Y
CF Studies: Gene Therapy
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CF Studies: First Imaging
-IMAGER
Image Autoradiograph
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CF Studies: Separate Imaging
Small animal imager with a highsensitivity copper collimator
Control with NaI Targeted delivery
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CF Study: Validation
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Reporter Gene
Suicide gene: HSV1-tk
HSV1-tk (gene) HSV1-TK (enzyme)
Ganciclovir (GCV) GCV-PO4
(cellular enzymes)
di/tri-PO4
Reporter gene: HSV1-tk Reporter gene probe: GCV
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Reporter Gene for Imaging (Imagene)
Reporter gene: HSV1-tk
HSV1-tk (gene) HSV1-TK (enzyme)
[125I]FIAU FIAU-PO4
(cellular enzymes)
di/tri-PO4
Radiolabeled FIAU or other probes trappedto image reporter gene expression in vivo
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TdR Analogues as Probe for HSV1-tk Expression
2’-fluoro-2’-deoxy-1--arabinofuranosyl-5-[I-125]-iodouracil (FIAU)
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CF Studies: Multimodal Imaging
Dual modality imaging
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CF Studies: Multimodal Imaging
Breathing mouse during imaging
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125I-FIAU Injection to Image Stable HSV1-tk Complexes
Artifacts De-halogenation
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HPLC Assay: [125I]-FIAU -trace
0.0 19.028 59.0
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X-ray detectoruniformitycorrection
Blockingthyroid uptake
Imaging with Corrections
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125I-FIAU Injection to Image Stable HSV1-tk Complexes
[I-125]-FIAU in Mouse Lungs
0
5
10
15
20
25
0.00 10.00 20.00 30.00 40.00 50.00 60.00 70.00 80.00 90.00
Time (post injection, hrs)
% o
f In
ject
ed D
ose
c1-both
c1-none
c4-both
c4-none
c5-right
c5-left
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Furture Plans
Multimodality Imaging for CF Gene TherapyBioluminescent Imaging of Luc
Cancer Imaging Projects Peptide Ligand (none-RGD) for angiogenesis
Imaging Inflammatory Response Fab targeting neutrophils
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Acknowledgement
Whitaker Foundation Imaging grant (RG-00-0305)
Cystic Fibrosis Foundation RDP grant (R477)
Department of Energy/JLab Subcontract (DE/SURA-04-Q011)
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CF Studies: uPET Imaging (UCLA)
tk + luc (compacted) saline tk (compated)
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CF Studies: uPET Imaging (UCLA)
microPET-measured sr39tk Expression in the Lung of Mice
0
0.5
1
1.5
2
0 1 2 3 4 5 6
Days (gene delivery at Day 0)
FH
BG
%ID
/g t
issu
e
tk(naked)
tk+luc(comp)
saline
tk(comp)
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Two Concepts
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Imaging Modalities
Anatomic Physiologic Metabolic Molecular
Nuclear (PET/SPECT)
CT
US
Optical Imaging
MR (MRI, MRS)
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PET/SPECT Radio-nuclides