Imaging expression of adenoviral HSV1-tk suicide gene transfer using the nucleoside analogue FIRU
| Autor: | Edward E. Knaus, Peter H. Cox, Leonard I. Wiebe, Ebrahim Naimi, Cees C J Avezaat, Peter A. E. Sillevis Smitt, M.J. Driesse, Magda Bijster, Kevin W. Morin, Ronald Vogels, Marion de Jong, Dharmin Nanda, Willem H. Bakker, Menzo J. E. Havenga |
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| Přispěvatelé: | Neurology, Radiology & Nuclear Medicine, Neurosurgery |
| Rok vydání: | 2002 |
| Předmět: |
Gene Expression Regulation
Viral Biodistribution Genetic Vectors Gene Expression Herpesvirus 1 Human Fialuridine Transfection Sensitivity and Specificity Adenoviridae Iodine Radioisotopes Mice Multiplicity of infection SDG 3 - Good Health and Well-being Genes Reporter Transduction Genetic In vivo Biomarkers Tumor Tumor Cells Cultured medicine Animals Humans Tissue Distribution Radiology Nuclear Medicine and imaging Radionuclide Imaging Onderzoek Neurochirurgie Chemistry Arabinofuranosyluracil Genetic transfer Gene Transfer Techniques Nucleosides Genetic Therapy Glioma General Medicine Suicide gene Molecular biology Cell culture Immunology Female Radiopharmaceuticals medicine.drug |
| Zdroj: | European Journal of Nuclear Medicine and Molecular Imaging, 29, 7, pp. 939-47 European Journal of Nuclear Medicine, 29(7), 939-947. Springer-Verlag European Journal of Nuclear Medicine and Molecular Imaging, 29, 939-47 |
| ISSN: | 1619-7070 0340-6997 |
| Popis: | Substrates for monitoring HSV1-tk gene expression include uracil and acycloguanosine derivatives.The most commonly used uracil derivative to monitor HSV1-tk gene transfer is 1-(2-fluoro-2-deoxy-β-D-arabinofuranosyl)-5-[*I]iodouracil (fialuridine; I*-FIAU), where the asterisk denotes any of the radioactive iodine isotopes that can be used. We have previously studied other nucleosides with imaging properties as good as or better than FIAU, including 1-(2-fluoro-2-deoxy-β-D-ribofuranosyl)-5-[*I]iodouracil (FIRU). The first aim of this study was to extend the biodistribution data of 123I-labelled FIRU. Secondly, we assessed the feasibility of detecting differences in HSV1-tk gene expression levels following adenoviral gene transfer in vivo with 123I-FIRU. 9L rat gliosarcoma cells were stably transfected with the HSV1-tk gene (9L-tk+). 123I-FIRU was prepared by radioiodination of 1-(2-fluoro-2-deoxy-β-D-ribofuranosyl)-5-tributylstannyl uracil (FTMRSU; precursor compound) and purified using an activated Sep-Pak column. Incubation of 9L-tk+ cells and the parental 9L cells with 123I-FIRU resulted in a 100-fold higher accumulation of radioactivity in the 9L-tk+ cells after an optimum incubation time of 4 h. NIH-bg-nu-xid mice were then inoculated subcutaneously with HSV1-tk (–) 9L cells or HSV1-tk (+) 9L-tk+ cells into both flanks. Biodistribution studies and gamma camera imaging were performed at 15 min and 1, 2, 4 and 24 h p.i. At 15 min, the tumour/muscle, tumour/blood and tumour/brain ratios were 5.2, 1.0 and 30.3 respectively. Rapid renal clearance of the tracer from the body resulted in increasing tumour/muscle, tumour/blood and tumour/brain ratios, reaching values of 32.2, 12.5 and 171.6 at 4 h p.i. A maximum specific activity of 22%ID/g tissue was reached in the 9L-tk+ tumours 4 h after 123I-FIRU injection. Two Ad5-based adenoviral vectors containing the HSV1-tk gene were constructed: a replication-incompetent vector with the transgene in the former E1 region, driven by a modified CMV promoter, and a novel replication-competent vector with the HSV1-tk gene in E3 driven by the natural E3 promoter. The human glioma cell lines U87MG and T98G were infected with a multiplicity of infection (m.o.i.) of 10. Forty-eight hours later the cells were incubated with 123I-FIRU and radioactivity was measured in a gamma counter. We found significantly higher levels of radioactivity in both cell lines following infection with the replication-competent vector (P |
| Databáze: | OpenAIRE |
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