Transfer of the cytidine deaminase cDNA into hematopoietic cells

Flaßhove, Michael; Frings, Werner; Schröder, J; Möritz, Thomas; Schütte, J.; Seeber, S.

doi:10.1016/s0145-2126(99)00128-9

Cited by 27 publications

(21 citation statements)

References 33 publications

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“…The degree of resistance to cytarabine or gemcitabine which we observed in human hematopoietic cells was comparable to that already described in hematopoetic cell lines or primary murine hematopoietic cells. Here, in general, resistance to cytarabine doses ranging from 10 to 500 nM 11,25 and gemcitabine doses ranging from 1 to 10 nM 9 has been reported, with the only exception of early work from Momparler et al 12 reporting resistance of CFU-C to cytarabine doses of up to 50 mM. However, also this group later described resistance in the aforementioned dose range only.…”

Section: Discussionmentioning

confidence: 87%

“…Consequently, CDD has been considered a potential drug resistance gene and this capacity has been demonstrated in cell lines and in the murine hematopoietic system. [7][8][9][10][11][12] To our knowledge, our study for the first time transfers these data to primary human hematopoietic cells. The degree of resistance to cytarabine or gemcitabine which we observed in human hematopoietic cells was comparable to that already described in hematopoetic cell lines or primary murine hematopoietic cells.…”

Section: Discussionmentioning

confidence: 99%

“…One obvious way to increase resistance levels would be better transgene expression. While vectors employing the SFFV/MESV promotor/enhancer activity have given excellent results with other therapeutic and marker genes in our as well as other investigators' hands, 11,17,18,23,29,[31][32][33] retroviral expression of transgenes from a specific retroviral construct has repeatedly been demonstrated to be transgene specific. Thus, the degree of hematoprotection achieved by CDD expression might be increased not only by the use of new and optimized retroviral vector backbones but also by using other vector systems such as lenti-or foamyvirus-based constructs.…”

Section: Discussionmentioning

confidence: 99%

“…[7][8][9] In addition, we and others have shown increased cytarabine resistance after retrovirally mediated transduction of primary murine hematopoietic cells. [10][11][12] More recently, increased gemcitabine and 5-aza-2 0 -deoxycytidine resistance after CDD gene transfer into murine cells 13,14 and transfer of the CDD gene into primary murine bone marrow stromal cells, followed by in vitro enrichment of these cells by cytarabine application, have been described. 14 Successful treatment of lymphoma with a combination of methotrexate and cytarabine in NOD/SCID mice transplanted with drug-resistant bone marrow could also be demonstrated.…”

Section: Introductionmentioning

confidence: 99%

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Resistance to cytarabine and gemcitabine and in vitro selection of transduced cells after retroviral expression of cytidine deaminase in human hematopoietic progenitor cells

et al. 2005

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Overexpression of the detoxifying enzyme cytidine deaminase (CDD) renders normal and leukemic hematopoietic cells resistant to cytarabine (1-b-D-arabinofuranosylcytosine), and studies on murine cells have suggested transgenic CDD overexpression as a way to reduce the substantial myelotoxicity induced by the deoxycytidine analogs cytarabine and gemcitabine (2 0 ,2 0 -difluorodeoxycytidine). We now have investigated CDD (over-)expression in the human hematopoietic system. Retroviral gene transfer significantly increased the resistance of CDD-transduced cord blood and peripheral bloodderived progenitor cells for doses ranging from 20 to 100 nM cytarabine and 8-10 nM gemcitabine. Protection was observed for progenitors of erythroid as well as myeloid differentiation, though the degree of protection varied for individual drugs. In addition, significant selection of CDD-transduced cells was obtained after a 4-day culture in 30-100 nM cytarabine. Thus, our data demonstrate that overexpression of CDD cDNA results in significant protection of human progenitors from cytarabineas well as gemcitabine-induced toxicity, and allows in vitro selection of transduced cells. This strongly argues for a potential therapeutic role of CDD gene transfer in conjunction with dose-intensive cytarabine-or gemcitabine-containing chemotherapy regimen.

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Section: Discussionmentioning

confidence: 87%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Resistance to cytarabine and gemcitabine and in vitro selection of transduced cells after retroviral expression of cytidine deaminase in human hematopoietic progenitor cells

et al. 2005

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“…This result was confirmed by others, both on fibroblasts and mouse bone marrow with araC, gemcitabine and 5-aza-deoxycytidine. [68][69][70][71][72] It was also shown that the resistance was reversed by using THU. 70 In the next step, transduced hematopoietic cells were transplanted into the bone marrow of mice to show the production of CDA in blood and different tissues.…”

Section: Myeloprotection With Cytidine Deaminasementioning

confidence: 99%

Development of gene therapy in association with clinically used cytotoxic deoxynucleoside analogues

2009

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The clinical use of cytotoxic deoxynucleoside analogues is often limited by resistance mechanisms due to enzymatic deficiency, or high toxicity in nontumor tissues. To improve the use of these drugs, gene therapy approaches have been proposed and studied, associating clinically used deoxynucleoside analogues such as araC and gemcitabine and suicide genes or myeloprotective genes. In this review, we provide an update of recent results in this area, with particular emphasis on human deoxycytidine kinase, the deoxyribonucleoside kinase from Drosophila melanogaster, purine nucleoside phosphorylase from Escherichia coli, and human cytidine deaminase. Data from literature clearly show the feasibility of these systems, and clinical trials are warranted to conclude on their use in the treatment of cancer patients.

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Improving the biosafety of cell sorting by adaptation of a cell sorting system to a biosafety cabinet

Lennartz

Flaßhove

et al. 2005

Cytometry Pt A

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Background: The jet-in-air cell sorters currently available are not very suitable for sorting potentially biohazardous material under optimal conditions because they do not protect operators and samples as recommended in the guidelines for safe biotechnology. To solve this problem we have adapted a cell sorting system to a special biosafety cabinet that satisfies the requirements for class II cabinets. With aid of this unit, sorting can be performed in conformance with the recommendations for biosafety level 2. Methods: After integrating a modified fluorescence-activated cell sorter (FACS) Vantage into a special biosafety cabinet, we investigated the influence of the laminar air flow (LAF) inside the cabinet on side stream stability and the analytical precision of the cell sorter. In addition to the routine electronic counting of microparticles, we carried out tests on the containment of aerosols, using T4 bacteriophage as indicators, to demonstrate the efficiency of the biosafety cabinet for sorting experiments performed under biosafety level 2 conditions. Results: The experiments showed that LAF, which is necessary to build up sterile conditions in a biosafety cabi-

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Transfer of the cytidine deaminase cDNA into hematopoietic cells

Cited by 27 publications

References 33 publications

Resistance to cytarabine and gemcitabine and in vitro selection of transduced cells after retroviral expression of cytidine deaminase in human hematopoietic progenitor cells

Resistance to cytarabine and gemcitabine and in vitro selection of transduced cells after retroviral expression of cytidine deaminase in human hematopoietic progenitor cells

Development of gene therapy in association with clinically used cytotoxic deoxynucleoside analogues

Improving the biosafety of cell sorting by adaptation of a cell sorting system to a biosafety cabinet

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