Imaging Expression of Cytosine Deaminase-Herpes Virus Thymidine Kinase Fusion Gene (CD/TK) Expression with [I]FIAU and PET
Double prodrug activation gene therapy using the Escherichia coli cytosine deaminase (CD)herpes simplex virus type 1 thymidine kinase ( HSV1-tk ) fusion gene ( CD/TK ) with 5-fluorocytosine (5FC), ganciclovir (GCV), and radiotherapy is currently under evaluation for treatment of different tumors. We...
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| Format: | Article |
| Language: | English |
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SAGE Publishing
2002-01-01
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| Series: | Molecular Imaging |
| Online Access: | https://doi.org/10.1162/15353500200200003 |
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| author | Trevor Hackman Michail Doubrovin Julius Balatoni Tatiana Beresten Vladimir Ponomarev Bradly Beattie Ronald Finn William Bornmann Ronald Blasberg Juri Gelovani Tjuvajev |
| author_facet | Trevor Hackman Michail Doubrovin Julius Balatoni Tatiana Beresten Vladimir Ponomarev Bradly Beattie Ronald Finn William Bornmann Ronald Blasberg Juri Gelovani Tjuvajev |
| author_sort | Trevor Hackman |
| collection | DOAJ |
| description | Double prodrug activation gene therapy using the Escherichia coli cytosine deaminase (CD)herpes simplex virus type 1 thymidine kinase ( HSV1-tk ) fusion gene ( CD/TK ) with 5-fluorocytosine (5FC), ganciclovir (GCV), and radiotherapy is currently under evaluation for treatment of different tumors. We assessed the efficacy of noninvasive imaging with [ 124 I]FIAU (2′-fluoro-2′-deoxy-1-β-d-arabinofuranosyl-5-iodo-uracil) and positron emission tomography (PET) for monitoring expression of the CD/TK fusion gene. Walker-256 tumor cells were transduced with a retroviral vector bearing the CD/TK gene (W256CD/TK cells). The activity of HSV1-TK and CD subunits of the CD/TK gene product was assessed in different single cell-derived clones of W256CD/TK cells using the FIAU radiotracer accumulation assay in cells and a CD enzyme assay in cell homogenates, respectively. A linear relationship was observed between the levels of CD and HSV1-tk subunit expression in corresponding clones in vitro over a wide range of CD/TK expression levels. Several clones of W256CD/TK cells with significantly different levels of CD/TK expression were selected and used to produce multiple subcutaneous tumors in rats. PET imaging of HSV1-TK subunit activity with [ 124 I]FIAU was performed on these animals and demonstrated that different levels of CD/TK expression in subcutaneous W256CD/TK tumors can be imaged quantitatively. CD expression in subcutaneous tumor sample homogenates was measured using a CD enzyme assay. A comparison of CD and HSV1-TK subunit enzymatic activity of the CD/TK fusion protein in vivo showed a significant correlation. Knowing this relationship, the parametric images of CD subunit activity were generated. Imaging with [ 124 I]FIAU and PET could provide pre- and posttreatment assessments of CD/TK -based double prodrug activation in clinical gene therapy trials. |
| format | Article |
| id | doaj-art-37c466b4fb1b404bb94006eec5e95014 |
| institution | Kabale University |
| issn | 1536-0121 |
| language | English |
| publishDate | 2002-01-01 |
| publisher | SAGE Publishing |
| record_format | Article |
| series | Molecular Imaging |
| spelling | doaj-art-37c466b4fb1b404bb94006eec5e950142025-01-02T22:38:07ZengSAGE PublishingMolecular Imaging1536-01212002-01-01110.1162/1535350020020000310.1162_15353500200200003Imaging Expression of Cytosine Deaminase-Herpes Virus Thymidine Kinase Fusion Gene (CD/TK) Expression with [I]FIAU and PETTrevor HackmanMichail DoubrovinJulius BalatoniTatiana BerestenVladimir PonomarevBradly BeattieRonald FinnWilliam BornmannRonald BlasbergJuri Gelovani TjuvajevDouble prodrug activation gene therapy using the Escherichia coli cytosine deaminase (CD)herpes simplex virus type 1 thymidine kinase ( HSV1-tk ) fusion gene ( CD/TK ) with 5-fluorocytosine (5FC), ganciclovir (GCV), and radiotherapy is currently under evaluation for treatment of different tumors. We assessed the efficacy of noninvasive imaging with [ 124 I]FIAU (2′-fluoro-2′-deoxy-1-β-d-arabinofuranosyl-5-iodo-uracil) and positron emission tomography (PET) for monitoring expression of the CD/TK fusion gene. Walker-256 tumor cells were transduced with a retroviral vector bearing the CD/TK gene (W256CD/TK cells). The activity of HSV1-TK and CD subunits of the CD/TK gene product was assessed in different single cell-derived clones of W256CD/TK cells using the FIAU radiotracer accumulation assay in cells and a CD enzyme assay in cell homogenates, respectively. A linear relationship was observed between the levels of CD and HSV1-tk subunit expression in corresponding clones in vitro over a wide range of CD/TK expression levels. Several clones of W256CD/TK cells with significantly different levels of CD/TK expression were selected and used to produce multiple subcutaneous tumors in rats. PET imaging of HSV1-TK subunit activity with [ 124 I]FIAU was performed on these animals and demonstrated that different levels of CD/TK expression in subcutaneous W256CD/TK tumors can be imaged quantitatively. CD expression in subcutaneous tumor sample homogenates was measured using a CD enzyme assay. A comparison of CD and HSV1-TK subunit enzymatic activity of the CD/TK fusion protein in vivo showed a significant correlation. Knowing this relationship, the parametric images of CD subunit activity were generated. Imaging with [ 124 I]FIAU and PET could provide pre- and posttreatment assessments of CD/TK -based double prodrug activation in clinical gene therapy trials.https://doi.org/10.1162/15353500200200003 |
| spellingShingle | Trevor Hackman Michail Doubrovin Julius Balatoni Tatiana Beresten Vladimir Ponomarev Bradly Beattie Ronald Finn William Bornmann Ronald Blasberg Juri Gelovani Tjuvajev Imaging Expression of Cytosine Deaminase-Herpes Virus Thymidine Kinase Fusion Gene (CD/TK) Expression with [I]FIAU and PET Molecular Imaging |
| title | Imaging Expression of Cytosine Deaminase-Herpes Virus Thymidine Kinase Fusion Gene (CD/TK) Expression with [I]FIAU and PET |
| title_full | Imaging Expression of Cytosine Deaminase-Herpes Virus Thymidine Kinase Fusion Gene (CD/TK) Expression with [I]FIAU and PET |
| title_fullStr | Imaging Expression of Cytosine Deaminase-Herpes Virus Thymidine Kinase Fusion Gene (CD/TK) Expression with [I]FIAU and PET |
| title_full_unstemmed | Imaging Expression of Cytosine Deaminase-Herpes Virus Thymidine Kinase Fusion Gene (CD/TK) Expression with [I]FIAU and PET |
| title_short | Imaging Expression of Cytosine Deaminase-Herpes Virus Thymidine Kinase Fusion Gene (CD/TK) Expression with [I]FIAU and PET |
| title_sort | imaging expression of cytosine deaminase herpes virus thymidine kinase fusion gene cd tk expression with i fiau and pet |
| url | https://doi.org/10.1162/15353500200200003 |
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