Recombinant adenoviral vector containing tumor-specific l-plastin promoter fused to cytosine deaminase gene as a transcription unit: generation and functional test

2004 ◽  
Vol 27 (6) ◽  
pp. 633-639 ◽  
Author(s):  
Injae Chung ◽  
Albert B. Deisseroth
Keyword(s):  
Adenoviral Vector ◽  
Cytosine Deaminase ◽  
Transcription Unit ◽  
Functional Test ◽  
Recombinant Adenoviral Vector ◽  
Cytosine Deaminase Gene

scholarly journals Cytosine Deaminase Adenoviral Vector and 5-Fluorocytosine Selectively Reduce Breast Cancer Cells 1 Million-Fold When They Contaminate Hematopoietic Cells: A Potential Purging Method for Autologous Transplantation

Blood ◽  
1998 ◽  
Vol 92 (2) ◽  
pp. 672-682 ◽  
Author(s):  
F. Garcia-Sanchez ◽  
G. Pizzorno ◽  
S.Q. Fu ◽  
T. Nanakorn ◽  
D.S. Krause ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Cancer Patients ◽  
Adenoviral Vector ◽  
Breast Cancer Cells ◽  
Hematopoietic Cells ◽  
Cytosine Deaminase ◽  
Transcription Unit ◽  
Breast Cancer Patients ◽  
Mcf 7

Abstract Ad.CMV-CD is a replication incompetent adenoviral vector carrying a cytomegalovirus (CMV)-driven transcription unit of the cytosine deaminase (CD) gene. The CD transcription unit in this vector catalyzes the deamination of the nontoxic pro-drug, 5-fluorocytosine (5-FC), thus converting it to the cytotoxic drug 5-fluorouracil (5-FU). This adenoviral vector prodrug activation system has been proposed for use in selectively sensitizing breast cancer cells, which may contaminate collections of autologous stem cells products from breast cancer patients, to the toxic effects of 5-FC, without damaging the reconstitutive capability of the normal hematopoietic cells. This system could conceivably kill even the nondividing breast cancer cells, because the levels of 5-FU generated by this system are 10 to 30 times that associated with systemic administration of 5-FU. The incorporation of 5-FU into mRNA at these high levels is sufficient to disrupt mRNA processing and protein synthesis so that even nondividing cells die of protein starvation. To test if the CD adenoviral vector sensitizes breast cancer cells to 5-FC, we exposed primary explants of normal human mammary epithelial cells (HMECs) and the established breast cancer cell (BCC) lines MCF-7 and MDA-MB-453 to the Ad.CMV-CD for 90 minutes. This produced a 100-fold sensitization of these epithelial cells to the effects of 48 hours of exposure to 5-FC. We next tested the selectivity of this system for BCC. When peripheral blood mononuclear cells (PBMCs), collected from cancer patients during the recovery phase from conventional dose chemotherapy-induced myelosuppression, were exposed to the Ad.CMV-CD for 90 minutes in serum-free conditions, little or no detectable conversion of 5-FC into 5-FU was seen even after 48 hours of exposure to high doses of 5-FC. In contrast, 70% of 5-FC was converted into the cytotoxic agent 5-FU when MCF-7 breast cancer cells (BCCs) were exposed to the same Ad.CMV-CD vector followed by 5-FC for 48 hours. All of the BCC lines tested were shown to be sensitive to infection by adenoviral vectors when exposed to a recombinant adenoviral vector containing the reporter gene betagalactosidase (Ad.CMV-βgal). In contrast, less than 1% of the CD34-selected cells and their more immature subsets, such as the CD34+CD38− or CD34+CD33− subpopulations, were positive for infection by the Ad.CMV-βgal vector, as judged by fluorescence-activated cell sorting (FACS) analysis, when exposed to the adenoviral vector under conditions that did not commit the early hematopoietic precursor cells to maturation. When artificial mixtures of hematopoietic cells and BCCs were exposed for 90 minutes to the Ad.CMV-CD vector and to 5-FC for 10 days or more, a greater than 1 million fold reduction in the number of BCCs, as measured by colony-limiting dilution assays, was observed. To test if the conditions were damaging for the hematopoietic reconstituting cells, marrow cells collected from 5-FU–treated male donor mice were incubated with the cytosine deaminase adenoviral vector and then exposed to 5-FC either for 4 days in vitro before transplantation or for 14 days immediately after transplantation in vivo. There was no significant decrease in the reconstituting capability of the male marrow cells, as measured by their persistence in female irradiated recipients for up to 6 months after transplantation. These observations suggest that adenovirus-mediated gene transfer of the Escherichia coli cytosine deaminase gene followed by exposure to the nontoxic pro-drug 5-FC may be a potential strategy to selectively reduce the level of contaminating BCCs in collections of hematopoietic cells used for autografts in breast cancer patients.

scholarly journals CBIO-40THREE-DIMENSIONAL ASSESSMENT OF BYSTANDER EFFECTS OF MESENCHYMAL STEM CELLS CARRYING A CYTOSINE DEAMINASE GENE ON GLIOMA CELLS

2015 ◽  
Vol 17 (suppl 5) ◽  
pp. v63.2-v63
Author(s):  
Jin Hwa Jung ◽  
Andrew Kim ◽  
Da-Young Chang ◽  
Sung-Soo Kim ◽  
Haeyoung Suh-Kim
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cytosine Deaminase ◽  
Glioma Cells ◽  
Bystander Effects ◽  
Dimensional Assessment ◽  
Cytosine Deaminase Gene

scholarly journals A Mutated Cytosine Deaminase Gene, codA (D314A), as an Efficient Negative Selection Marker for Gene Targeting in Rice

2014 ◽  
Vol 55 (3) ◽  
pp. 658-665 ◽  
Author(s):  
Keishi Osakabe ◽  
Ayako Nishizawa-Yokoi ◽  
Namie Ohtsuki ◽  
Yuriko Osakabe ◽  
Seiichi Toki
Keyword(s):  
Gene Targeting ◽  
Negative Selection ◽  
Cytosine Deaminase ◽  
Selection Marker ◽  
Negative Selection Marker ◽  
Cytosine Deaminase Gene
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