In vivo imaging of herpes simplex virus type 1 thymidine kinase gene expression: early kinetics of radiolabelled FIAU

Previous studies have shown that the herpes simplex virus type 1 thymidine kinase gene (HSV1-tk), in combination with appropriate radiolabelled substrates (e.g. [I*]-2'-fluoro-2'-deoxy-5-iodo-1-beta-D-arabinofuranosyluracil; I*-FIAU, where the asterisk indicates that any of the various radioactive iodine isotopes can be used), can be used as a reporter gene for in vivo monitoring of gene transfer and expression. The aim of our study was to examine the early kinetics of I*-FIAU and the possibility of utilising iodine-123-labelled FIAU for imaging of gene expression, CMS-5 fibrosarcoma cells were transduced in vitro with the retroviral vector STK containing the HSV1-tk gene. BALB/c mice were inoculated subcutaneously with HSV1-tk(+) and tk(-) cells into both flanks. FAU (2'-fluoro-2'-deoxy-1-beta-D-arabinofuranosyluracil was radioiodinated (I-123, I-125) using the iodogen method, High-performance liquid chromatography purification resulted in high specific activity and radiochemical purity for both tracers ([I-123]FIAU and [I-125]FIAU). Biodistribution studies and gamma camera imaging were performed at 0.5, 1, 2 and 4 h p.i. In addition, the genomic DNA of the tumours was isolated for measurement of the activity accumulation resulting from the [I-125]FIAU incorporation. Biodistribution studies 0.5 h p.i, showed tumour/blood and tumour/muscle ratios of 3.8 and 7.2, respectively, for the HSV1-tk(+) tumours, and 0.6 and 1.2, respectively, for negative control tumours, Fast renal elimination of the tracer from the body resulted in rapidly increasing tumour/blood and tumour/muscle ratios which reached values of 32 and 88 at 4 h p.i., respectively. Tracer clearance from blood was bi-exponential. with an initial half-life of 0.6 h followed by a half-life of 4.6 h, The tracer half-life in herpes simplex viral thymidine kinase-expressing tumours was 35.7 h. The highest activity accumulation (20.3%+/-5.7% ID/g) in HSV1-tk(+) tumours was observed 1 h p.i. At that time, about 46% of the total activity found in HSV1-tk(+) tumours was incorporated into genomic DNA, Planar gamma-camera imaging showed a distinct tracer accumulation as early as 0.5 h p.i., with an increase in contrast over time.-These results suggest that sufficient tumour/background ratios for in vivo imaging of HSV1-tk expression with [I-123]FIAU are reached as early as 1 h p.i.