Protection of hematopoietic stem cells from pemetrexed toxicity by retroviral gene transfer with a mutant dihydrofolate reductase-mutant thymidylate synthase fusion gene

Capiaux, Gina; Budak‐Alpdoğan, Tülin; Alpdoğan, Önder; Bornmann, William G.; Takebe, Naoko; Banerjee, Debabrata; Maley, Frank; Bertino, Joseph R.

doi:10.1038/sj.cgt.7700683

Cited by 14 publications

(6 citation statements)

References 24 publications

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“…A number of such CTX-R genes have been identified and for several of them protection of haematopoiesis from the associated cytotoxic agents has been established in murine as well as large animal models. 20,21 This includes mutant forms of dihydrofolate reductase (mutDHFR), [22][23][24] the multidrugresistance type 1 (MDR1) gene coding for the cellular efflux pump p-glycoprotein [25][26][27] or the gene coding for the DNA repair protein O 6 -methylguanine methyltransferase. [28][29][30][31] Cytidine deaminase represents another interesting CTX-R gene that protects lymphohaematopoietic cells from the cytotoxic deoxycytidine analogues cytosine-arabinoside (1-b-D-arabinofuranosylcytosine, Ara-C), gemcitabine (2 0 ,2 0 -difluorodeoxycytidine) and 5-azacitabine (5-aza-2 0 deoxycytidine).…”

Section: Introductionmentioning

confidence: 99%

Efficient in vivo regulation of cytidine deaminase expression in the haematopoietic system using a doxycycline-inducible lentiviral vector system

et al. 2012

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Regulated transgene expression may reduce transgene-specific and genotoxic risks associated with gene therapy. To prove this concept, we have investigated the suitability of doxycycline (Dox)-inducible human cytidine deaminase (hCDD) overexpression from lentiviral vectors to mediate effective myeloprotection while circumventing the lymphotoxicity observed with constitutive CDD activity. Rapid Dox-mediated transgene induction associated with a 6-17-fold increase in drug resistance was observed in 32D and primary murine bone marrow (BM) cells. Moreover, robust Dox-regulated transgene expression in the entire haematopoietic system was demonstrated for primary and secondary recipients of hCDD-transduced R26-M2rtTA transgenic BM cells. Furthermore, mice were significantly protected from myelosuppressive chemotherapy as evidenced by accelerated recovery of granulocytes (1.9±0.6 vs 1.3±0.3, P=0.034) and platelets (883±194 vs 584±160 10(3) per μl, P=0.011). Minimal transgene expression in the non-induced state and no overt cellular toxicities including lymphotoxicity were detected. Thus, using a relevant murine transplant model our data provide conclusive evidence that drug-resistance transgenes can be expressed in a regulated fashion in the lymphohaematopoietic system, and that Dox-inducible systems may be used to reduce myelotoxic side effect of anticancer chemotherapy or to avoid side effects of high constitutive transgene expression.

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Section: Introductionmentioning

confidence: 99%

Efficient in vivo regulation of cytidine deaminase expression in the haematopoietic system using a doxycycline-inducible lentiviral vector system

et al. 2012

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“…been used to transfer MTX-resistant hDHFRs into mouse and human bone marrow progenitor cells, efficiently ensuring stem cell survival upon exposure to MTX. 25,26 Resistant cells transplanted into the bone marrow of mice ensured myeloprotection during treatment with MTX. 27,28 For this application, ideal candidate hDHFR mutants should have a very high K i for MTX (in the high nanomolar range), while maintaining the catalytic properties, including DHF binding, required to ensure cell survival.…”

mentioning

confidence: 99%

Increasing Methotrexate Resistance by Combination of Active-site Mutations in Human Dihydrofolate Reductase

Volpato

Fossati

Pelletier

2007

Journal of Molecular Biology

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“…An advantage of DHFR FS is that it can be used to select for transgenes useful for therapeutic efficacy such as tumor targeting proteins such as chimeric antigen receptors (8), suicide genes (9), or imaging genes (10). In an effort to broaden the application of DHFR FS , our group added a human TYMS mutant, discovered in a bacterial screen, resistant to 5-fluorouracil (5-FU) (11 (12,13). We suggested that the addition of TYMS SS restores thymidine synthesis in MTX-treated T cells (9).…”

Section: Methotrexate (Mtx) Is An Anti-folate That Inhibits De Novo Pmentioning

confidence: 99%

Dihydrofolate Reductase and Thymidylate Synthase Transgenes Resistant to Methotrexate Interact to Permit Novel Transgene Regulation

Rushworth

Mathews

Alpert

et al. 2015

Journal of Biological Chemistry

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Background:Methotrexate targets dihydrofolate reductase (DHFR) and thymidylate synthase (TYMS) to prevent cancer growth, and increases DHFR expression when applied. Results: TYMS resistant to methotrexate inhibits the methotrexate induced increase in DHFR expression. Conclusion: Thymidine synthesis regulates post-transcriptional expression of DHFR and TYMS. Significance: Methotrexate-resistant DHFR and TYMS can be used to regulate cis and trans transgene in primary T cells.

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Protection of hematopoietic stem cells from pemetrexed toxicity by retroviral gene transfer with a mutant dihydrofolate reductase-mutant thymidylate synthase fusion gene

Cited by 14 publications

References 24 publications

Efficient in vivo regulation of cytidine deaminase expression in the haematopoietic system using a doxycycline-inducible lentiviral vector system

Efficient in vivo regulation of cytidine deaminase expression in the haematopoietic system using a doxycycline-inducible lentiviral vector system

Increasing Methotrexate Resistance by Combination of Active-site Mutations in Human Dihydrofolate Reductase

Dihydrofolate Reductase and Thymidylate Synthase Transgenes Resistant to Methotrexate Interact to Permit Novel Transgene Regulation

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