Cytosine deaminase and thymidine kinase gene therapy in a Dunning rat prostate tumour model: absence of bystander effects and characterisation of 5-fluorocytosine metabolism with 19F-NMR spectroscopy

Corban-Wilhelm, Heike; Hull, William E.; Becker, G.; Bauder-Wüst, Ulrike; Greulich, D; Debus, Jürgen

doi:10.1038/sj.gt.3301834

Cited by 23 publications

(23 citation statements)

References 32 publications

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“…The levels of nascent pre-mRNA were not influenced by exposure to 5-FU, but mature mRNA declined by relatively modest concentrations of 0.13 mg/ml 5-FU. 22 In another experiment, 23 we incubated AT-1 and AT-1/CDglyTK cells for 24 hours with various concentrations of 5-FU (IC 50(MTS) ¼ 0.4 mg/ml), and then cultured further with fresh medium. Both cell lines showed essentially the same behavior.…”

Section: Comparison Of Different Toxicological Methods H Corban-wilhementioning

confidence: 99%

Comparison of different methods to assess the cytotoxic effects of cytosine deaminase and thymidine kinase gene therapy

et al. 2003

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Dunning R3327 AT-1 rat prostate tumor cells were transfected with a double-fusion suicide gene (CDglyTK) that coded for the cytosine deaminase from E. coli and the thymidine kinase (TK) from HSV-1. The resulting cell line AT-1/CDglyTK was incubated with 10 and 20 mg/ml 5-FC or 0.25 mg/ml GCV, or both 5-FC and GCV 96 hours before harvest. The MTS assay detected cell viabilities of 5075 and 2575% after 5-FC treatment, and 5075% after GCV treatment. The dye exclusion and the colony-forming assay confirmed the data of the MTS assay with GCV (4775 and 3275%), but presented different results for the 5-FC incubation. We detected 10071 and 8575% viable cells after 10 mg/ml 5-FC, and 9771 and 8575% after 20 mg/ml 5-FC treatment, respectively. S-phase arrest in both suicide gene systems was noticeable and a significant increase in cell granularity was observed after incubation with GCV or GCV & 5-FC. This study demonstrates that 5-FC and the metabolized 5-FU act not only as genotoxic reagents, but also as RNA-directed agent, because of the recovery of the cells. On the other hand, a significant S-phase block could be observed after 24 hours incubation with GCV. This short time is enough to incorporate the genotoxic GCV metabolites in the nascent DNA to impair the cell cycle.

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Section: Comparison Of Different Toxicological Methods H Corban-wilhementioning

confidence: 99%

Comparison of different methods to assess the cytotoxic effects of cytosine deaminase and thymidine kinase gene therapy

et al. 2003

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“…The results indicated that both yCDglyTK/5-FC and yCDglyTK/GCV were active, and the cytotoxicity of yCDglyTK/5-FC sysytem is higher than that of yCDglyTK/GCV for NPC cells, but the combined system did not show the optimal effect among the three systems that were of our expectations, but superior to yCDglyTK/ GCV system and inferior to the yCDglyTK/5-FC system, which differed from other reports. 6,29,30 These observations could be explained in two ways. First, the yCD and TK may have reciprocal interference in our yCDglyTK/5-FC þ GCV system for cancer therapy.…”

Section: Discussionmentioning

confidence: 99%

A novel fusion suicide gene yeast CDglyTK plays a role in radio-gene therapy of nasopharyngeal carcinoma

et al. 2004

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To investigate a novel suicide gene for nasopharyngeal carcinoma (NPC) therapy, the yCDglyTK gene was constructed by fusing yeast cytosine deaminase (CD) and herpes simplex type 1 thymidine kinase. The expression of the yCDglyTK gene was detected by RT-PCR and Western blotting, and its bioactivity was demonstrated by an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay. An animal study was carried out in which BALB/C nude mice bearing yCDglyTK gene-modified tumors were treated with prodrugs and radiation. Our results revealed that the yCDglyTK gene could be expressed in CNE-2 cells in vitro. In MTT analysis, at the transfection rate of 10%, 66% cells were killed. The synergistic effect of CD and TK showed 91% of yCDglyTK-transfected cells were killed with the treatment of 5-fluorocytosine (5-FC) alone, 60% killed with ganciclovir (GCV) alone, and 75% killed with 5-FC and GCV together. In vivo, the tumor volume in all of the four prodrugs and/or radiation-treated groups were significantly different from that in the PBS-controlled group (Po.01); also yCDglyTK þ prodrug þ radiation group was different from the other three groups (Po.05). Our findings suggested there was a synergistic antitumor effect when combining suicide gene therapy and radiation, and yCDglyTK has potent antitumor efficacy and may be a candidate suicide gene for cancer therapy.

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“…CD activates the minimally toxic 5-fluorocytosine (5FC) to the highly toxic 5-fluorouracil (5FU) [146,147]. The conversion of 5FC to 5FU causes a 19 F NMR chemical shift approximately 1.5 ppm, hence, revealing gene activity, which has been demonstrated in a number of systems in vivo [147,150].…”

Section: Enzyme Reportersmentioning

confidence: 99%

“…Given the inherent dose-limiting toxicity of 5FU, various prodrugs and mixture formulations have been developed (e.g., capecitabine (Xeloda), Tegafur-uracil (Uftoral ® ), emitefur (3 (3-(6-benzoyloxy-3-cyano-2-pyridyloxycarbonyl)benzoyl)-1-ethoxymethyl-5-fluorouracil)) and 19 F NMR has played a role in analysis and development [16,63]. A new and potentially exciting application is assessment of prodrug therapy in conjunction with gene therapy; specifically, the use of cytosine deaminase (CD), to convert the relatively innocuous 5-fluorocytosine (5FC) to 5FU [146][147][148][149][150]. Several investigations have now reported 19 F NMR of the conversion of 5FC to 5FU based on the 19 F NMR chemical shift, Dd ¼ 2 ppm [18,147,150,151].…”

Section: U N C O R R E C T E D P R O O Fmentioning

confidence: 99%

VatuximabTM: Optimizing Therapeutic Strategies for Prostate Cancer Based on Dynamic MR Tumor Oximetry

Mason¹

2007

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Cytosine deaminase and thymidine kinase gene therapy in a Dunning rat prostate tumour model: absence of bystander effects and characterisation of 5-fluorocytosine metabolism with 19F-NMR spectroscopy

Cited by 23 publications

References 32 publications

Comparison of different methods to assess the cytotoxic effects of cytosine deaminase and thymidine kinase gene therapy

Comparison of different methods to assess the cytotoxic effects of cytosine deaminase and thymidine kinase gene therapy

A novel fusion suicide gene yeast CDglyTK plays a role in radio-gene therapy of nasopharyngeal carcinoma

VatuximabTM: Optimizing Therapeutic Strategies for Prostate Cancer Based on Dynamic MR Tumor Oximetry

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