Nucl Med Mol Imaging.  2009 Oct;43(5):468-477.

Imaging of Herpes Simplex Virus Type 1 Thymidine Kinase Gene Expression with Radiolabeled 5-(2-iodovinyl)-2'-deoxyuridine (IVDU) in Liver by Hydrodynamic-based Procedure

• Song I
• Lee TS
• Kang JH
• Lee YJ
• Kim KI
• An GI
• Chung WS
• Cheon GJ
• Choi CW
• Lim SM

Abstract

PURPOSE
Hydrodynamic-based procedure is a simple and effective gene delivery method to lead a high gene expression in liver tissue. Non-invasive imaging reporter gene system has been used widely with herpes simplex virus type 1 thymidine kinase (HSV1-tk) and its various substrates. In the present study, we investigated to image the expression of HSV1-tk gene with 5-(2-iodovinyl)-2'-deoxyuridine (IVDU) in mouse liver by the hydrodynamicbased procedure.
MATERIALS AND METHODS
HSV1-tk or enhanced green fluorescence protein (EGFP) encoded plasmid DNA was transferred into the mouse liver by hydrodynaminc injection. At 24 h post-injection, RT-PCR, biodistribution, fluorescence imaging, nuclear imaging and digital wholebody autoradiography (DWBA) were performed to confirm transferred gene expression.
RESULTS
In RT-PCR assay using mRNA from the mouse liver, specific bands of HSV1-tk and EGFP gene were observed in HSV1-tk and EGFP expressing plasmid injected mouse, respectively. Higher uptake of radiolabeled IVDU was exhibited in liver of HSV1-tk gene transferred mouse by biodistribution study. In fluorescence imaging, the liver showed specific fluorescence signal in EGFP gene transferred mouse. Gamma-camera image and DWBA results showed that radiolabeled IVDU was accumulated in the liver of HSV1-tk gene transferred mouse.
CONCLUSION
In this study, hydrodynamic-based procedure was effective in liver-specific gene delivery and it could be quantified with molecular imaging methods. Therefore, co-expression of HSV1-tk reporter gene and target gene by hydrodynamic-based procedure is expected to be a useful method for the evaluation of the target gene expression level with radiolabeled IVDU.

Keyword

Hydrodynamic-based procedure; HSV1-tk; IVDU; liver; molecular imaging

MeSH Terms

Animals
Autoradiography
DNA
Fluorescence
Gene Expression
Genes, Reporter
Herpes Simplex
Herpesvirus 1, Human
Idoxuridine
Liver
Methylmethacrylates
Mice
Molecular Imaging
Optical Imaging
Plasmids
Polystyrenes
RNA, Messenger
Simplexvirus
Thymidine Kinase
DNA
Idoxuridine
Methylmethacrylates
Polystyrenes
RNA, Messenger
Thymidine Kinase