Drug selection of MDR1-transduced hematopoietic cells ex vivo increases transgene expression and chemoresistance in reconstituted bone marrow in mice

Licht, Thomas; Goldenberg, Sarah; Vieira, Wilfred D.; Gottesman, Michael M.; Pastan, Ira

doi:10.1038/sj.gt.3301087

Cited by 23 publications

(16 citation statements)

References 47 publications

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“…Ex vivo selection of BMCs with MDR drugs also produced elevated transgene expression and showed increase chemoresistance in reconstituted bone marrow of mice. 37,38 We can conclude that the human CD gene is able to chemoprotect murine BMCs against diverse deoxycytidine analogs. This protection is high enough to allow excellent in vitro selection potential.…”

Section: Discussionmentioning

confidence: 99%

Selection of drug-resistant transduced cells with cytosine nucleoside analogs using the human cytidine deaminase gene

et al. 2001

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Hematopoietic toxicity produced by most anticancer drugs limits their potential for curative therapy. We have shown previously that the human cytidine deaminase ( CD ) gene can confer drug resistance in murine bone marrow cells ( BMCs ) to the nucleoside analog, cytosine arabinoside ( ARA -C ) . In the present study, as the first objective we showed that the CD gene can also render drug resistance in BMCs to related analogs, 2 0 ,2 0 -difluorodeoxycytidine ( dFdC ) and 5 -azadeoxycytidine ( 5 -AZA -CdR ) . As a second objective, we investigated the potential of ex vivo selection with cytosine nucleoside analogs of CD -transduced BMC. The goal of this approach was to enrich the fraction of CD -transduced BMCs so as to increase the transgene expression and level of drug resistance before transplantation. This strategy may have the potential to circumvent the problem in clinical gene therapy of low level of gene transfer and adequate long -term gene expression. Using a bicistronic retroviral vector containing the CD and the green fluorescent protein ( CDiGFP ) , we transduced murine L1210 leukemic cells. All three analogs, ARA -C, dFdC, and 5 -AZA -CdR were demonstrated in vitro to enrich ( > 95% ) the population of leukemic cells expressing the GFP transgene. However, with CD -transduced primary murine BMCs cultivated at high cell density we observed that in vitro selection with ARA -C was not possible due to release of CD into the culture medium at amounts that were sufficient to inactivate the analog. The CD -containing medium produced a chemoprotective effect on mock BMCs as shown by lack of significant growth inhibition in the presence of ARA -C. However, at low cell density in a cell mixture containing CD -transduced cells, the mock BMCs showed marked drug sensitivity to ARA -C as determined by clonogenic assay. Selection with ARA -C was shown to significantly increase the CD enzyme activity in transduced BMC. These results suggest that CD gene has the potential to be a good selectable marker and a possible tool for chemoprotection in cancer gene therapy. Cancer Gene Therapy ( 2001 ) 8, 669 -676

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Section: Discussionmentioning

confidence: 99%

Selection of drug-resistant transduced cells with cytosine nucleoside analogs using the human cytidine deaminase gene

et al. 2001

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“…Additionally, selection will maintain expression of the therapeutic gene at a higher level for long time periods. Previously, MDR has been used successfully as a selectable marker to target hematopoietic progenitor cells (Aran et al, 1996;Galipeau et al, 1997;Hildinger et al, 1998;Licht et al, 1999;Licht et al, 2000;Sugimoto et al, 1995;Sugimoto et al, 2003;Zhou et al, 1998). We have developed a bicistronic retroviral vector expressing atrial natriuretic peptide (ANP) and MDR in a genetically modified HSE.…”

Section: Model For Systemic Delivery Of Therapeutic Proteinsmentioning

confidence: 99%

An Approach to Achieve Long-Term Expression in Skin Gene Therapy

2008

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For gene therapy purposes, the skin is an attractive organ to target for systemic delivery of therapeutic proteins to treat systemic diseases, skin diseases, or skin cancer. To achieve long-term stable expression of a therapeutic gene in keratinocytes (KC), we have developed an approach using a bicistronic retroviral vector expressing the desired therapeutic gene linked to a selectable marker (multidrug resistant gene, MDR) that is then introduced into KC and fibroblasts (FB) to create genetically modified human skin equivalent (HSE). After grafting the HSE onto immunocompromised mice, topical colchicine treatment is used to select and enrich for genetically modified keratinocyte stem cells (KSC) that express MDR and are resistant to colchicine's antimitotic effects. Both the apparatus for topical colchicine delivery and the colchicine doses have been optimized for application to human skin. This approach can be validated by systemic delivery of therapeutic factors such as erythropoietin and the antihypertensive atrial natriuretic peptide.

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“…Certain drug resistance genes, for example, the multidrug resistance gene 1 (MDR1), or multidrug resistance-associated protein 1 (MRP1) gene could provide resistance to a variety of chemotherapeutics that could be used in combination. 1,82,85,86,[99][100][101][102] Expressing more than one drug resistance gene to protect against a combination of chemotherapeutic drugs could be of great therapeutic benefit. These genes may be expressed from a single promoter in order to circumvent the issues of different transcriptional efficiencies of different promoters or may be expressed from the same promoter but as separate translational units by the use of internal ribosomal entry site (IRES) elements.…”

Section: Drug Resistance Gene Transfer For Myeloprotectionmentioning

confidence: 99%

“…1, [73][74][75][76][77][78][79][80][81][82][83][84][85] Myeloprotection-based gene therapy strategies have led to many preclinical studies 81,84,[86][87][88][89][90][91][92][93][94][95][96][97] and a few clinical studies.…”

Section: Drug Resistance Gene Transfer For Myeloprotectionmentioning

confidence: 99%

Hematopoietic stem cell gene therapy with drug resistance genes: an update

2005

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Transfer of drug resistance genes into hematopoietic stem cells (HSCs) has promise for the treatment of a variety of inherited, that is, X-linked severe combined immune deficiency, adenosine deaminase deficiency, thalassemia, and acquired disorders, that is, breast cancer, lymphomas, brain tumors, and testicular cancer. Drug resistance genes are transferred into HSCs either for providing myeloprotection against chemotherapy-induced myelosuppression or for selecting HSCs that are concomitantly transduced with another gene for correction of an inherited disorder. In this review, we describe ongoing experimental approaches, observations from clinical trials, and safety concerns related to the drug resistance gene transfer.

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Drug selection of MDR1-transduced hematopoietic cells ex vivo increases transgene expression and chemoresistance in reconstituted bone marrow in mice

Cited by 23 publications

References 47 publications

Selection of drug-resistant transduced cells with cytosine nucleoside analogs using the human cytidine deaminase gene

Selection of drug-resistant transduced cells with cytosine nucleoside analogs using the human cytidine deaminase gene

An Approach to Achieve Long-Term Expression in Skin Gene Therapy

Hematopoietic stem cell gene therapy with drug resistance genes: an update

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