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.

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.

Molecular/Small Animal Imaging

Planar Multimodal Imaging System

Small animal scanner,developed by JLab at

CWRU/UHC, combining x-ray and scintigraphy

CF Gene Therapy Measurement

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

CF Studies: First Imaging

-IMAGER

Image Autoradiograph

CF Studies: Separate Imaging

Small animal imager with a highsensitivity copper collimator

Control with NaI Targeted delivery

CF Study: Validation

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

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

TdR Analogues as Probe for HSV1-tk Expression

2’-fluoro-2’-deoxy-1--arabinofuranosyl-5-[I-125]-iodouracil (FIAU)

CF Studies: Multimodal Imaging

Dual modality imaging

CF Studies: Multimodal Imaging

Breathing mouse during imaging

125I-FIAU Injection to Image Stable HSV1-tk Complexes

Artifacts De-halogenation

HPLC Assay: [125I]-FIAU -trace

0.0 19.028 59.0

X-ray detectoruniformitycorrection

Blockingthyroid uptake

Imaging with Corrections

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

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

Acknowledgement

Whitaker Foundation Imaging grant (RG-00-0305)

Cystic Fibrosis Foundation RDP grant (R477)

Department of Energy/JLab Subcontract (DE/SURA-04-Q011)

CF Studies: uPET Imaging (UCLA)

tk + luc (compacted) saline tk (compated)

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)

Two Concepts

Imaging Modalities

Anatomic Physiologic Metabolic Molecular

Nuclear (PET/SPECT)

CT

US

Optical Imaging

MR (MRI, MRS)

PET/SPECT Radio-nuclides