alexa Comparison between adenoviral and retroviral vectors for the transduction of the thymidine kinase PET reporter gene in rat mesenchymal stem cells.
Genetics & Molecular Biology

Genetics & Molecular Biology

Journal of Genetic Syndromes & Gene Therapy

Author(s): Roelants V, Labar D, de Meester C, Havaux X, Tabilio A,

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Abstract Mesenchymal stem cells (MSCs) are a promising cell line for the treatment of ischemic heart disease. To evaluate the success of their transplantation into living animals, noninvasive imaging techniques that are able to track the distribution and fate of those cells would be useful. The aim of this study was to investigate the feasibility of infecting rat MSCs with adenoviruses and retroviruses carrying the herpes simplex virus type 1 thymidine kinase (HSV1-tk) gene; to compare the level of transgene expression induced by the 2 viral vectors; to evaluate the effects of viral transduction on cell phenotype, viability, proliferation rates, and differentiation capabilities; and to test the possibility of noninvasively imaging transduced MSCs using 9-(4-18F-fluoro-3-[hydroxymethyl]butyl)guanine (18F-FHBG) and small-animal PET after their transplantation into living rats. METHODS: We infected rat bone marrow MSCs with adenoviruses carrying the HSV1 mutant tk (Ad-HSV1-sr39tk) PET reporter gene (PRG) or with a retroviral construct expressing the wild-type HSV1-tk PRG. The efficacy and intensity of HSV1-sr39tk and HSV1-tk gene expression were determined by a direct comparison of [8-3H]-penciclovir ([8-3H]-PCV) cell uptake in both infected MSC populations and noninfected control MSCs. Small-animal PET studies were performed on living rats after an intramuscular injection of infected MSCs. The MSCs either have been incubated in advance with 18F-FHBG or they were administered and 18F-FHBG was thereafter intravenously administered [corrected] RESULTS: Both adenoviral and retroviral vectors can be used to introduce the tk PRG in MSCs. Neither adenovirus nor retrovirus infections significantly modify MSC phenotype, viability, proliferation, and differentiation capabilities. No significant 3H-PCV uptake was observed in noninfected MSCs. By contrast, after both adenoviral and retroviral infections, the infected MSC populations exhibited a similar, significantly higher, 3H-PCV accumulation. Small-animal PET images showed intense activity within the transplanted regions irrespective of the infected MSC population used. CONCLUSION: Our results demonstrate the feasibility of infecting MSCs with adenoviruses and retroviruses expressing the HSV1-tk PRG and suggest that infected MSCs can be noninvasively imaged with 18F-FHBG and small-animal PET after their transplantation into living animals. This article was published in J Nucl Med and referenced in Journal of Genetic Syndromes & Gene Therapy

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