Development of a retroviral construct containing a human mutated dihydrofolate reductase cDNA for hematopoietic stem cell transduction

@article{Li1994DevelopmentOA,
  title={Development of a retroviral construct containing a human mutated dihydrofolate reductase cDNA for hematopoietic stem cell transduction},
  author={Ming-xia Li and D. Banerjee and Shi-cheng Zhao and B. Schweitzer and S. Mineishi and E. Gilboa and J. Bertino},
  journal={Blood},
  year={1994},
  volume={83},
  pages={3403-3408}
}
A double-copy Moloney leukemia virus-based retroviral construct containing both the NeoR gene and a mutant human dihydrofolate reductase (DHFR) cDNA (Ser31 mutant) was used to transduce NIH 3T3 and mouse bone marrow (BM) progenitor cells. This resulted in increased resistance of these cells to methotrexate (MTX). The transduced BM progenitor cells were returned to lethally irradiated mice. The recipients transplanted with marrow cells infected with the recombinant virus showed protection from… Expand
Efficient protection from methotrexate toxicity and selection of transduced human hematopoietic cells following gene transfer of dihydrofolate reductase mutants.
TLDR
Gene transfer of DHFR using suitable retroviral backbones and DHFR mutants significantly increases MTX resistance of human CFU-C and allows efficient in vitro selection of transduced cells using a short-term selection procedure. Expand
CHAPTER 23 – Development and Application of an Engineered Dihydrofolate Reductase and Cytidine-Deaminase-Based Fusion Genes in Myeloprotection-Based Gene Therapy Strategies
TLDR
The results demonstrated both the feasibility and efficiency of ex vivo CD34 + stem cell transduction with this retroviral vector and in vivo selection in the presence of MTX. Expand
Comparison of methotrexate resistance conferred by a mutated dihydrofolate reductase (DHFR) cDNA in two different retroviral vectors
TLDR
The results indicate that the in vitro transduction efficiency and transgene expression of human DHFR in hematopoietic progenitor cells is equally conferred by both vectors. Expand
Retroviral transduction of human CD34+ umbilical cord blood progenitor cells with a mutated dihydrofolate reductase cDNA.
TLDR
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Coexpression of cytidine deaminase and mutant dihydrofolate reductase by a bicistronic retroviral vector confers resistance to cytosine arabinoside and methotrexate.
TLDR
The potential of the MFG-DHFR-IRES/CD vector to confer drug resistance to both MTX and ARA-C and may have future application in chemoprotection of normal hematopoietic cells in patients with cancer is demonstrated. Expand
Retroviral transduction of human dihydropyrimidine dehydrogenase cDNA confers resistance to 5-fluorouracil in murine hematopoietic progenitor cells and human CD34+-enriched peripheral blood progenitor cells
TLDR
It is demonstrated here that overexpression of human DPD confers resistance to 5-FU in NIH3T3 cells, mouse bone marrow cells, and in human CD34+-enriched hematopoietic progenitor cells, which encourages the use of this gene as a method to protect patients from 4-FU myelotoxicity. Expand
A gene transfer strategy for making bone marrow cells resistant to trimetrexate.
TLDR
It is concluded that the L22Y vector is highly effective in protecting hematopoiesis from T MTX toxicity and may provide a means for increasing the therapeutic utility of TMTX in certain cancers. Expand
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TLDR
The results demonstrate the effectiveness of lentivirus vectors for chemoprotection in a well developed animal model, with the potential for further preclinical development toward human application. Expand
Ex vivo expansion and selection of human CD34+ peripheral blood progenitor cells after introduction of a mutated dihydrofolate reductase cDNA via retroviral gene transfer.
TLDR
It is demonstrated that human hematopoietic precursor cells can be expanded extensively after retroviral gene transfer and the same population of early progenitors can be selected ex vivo with low-dose MTX. Expand
Retrovirus-mediated gene transfer of rat glutathione S-transferase Yc confers in vitro resistance to alkylating agents in human leukemia cells and in clonogenic mouse hematopoietic progenitor cells.
TLDR
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Chinese hamster ovary (CHO) DHFR- cells were converted into the DHFR+ phenotype when they were transfected with a mammalian expression vector carrying human dihydrofolate reductase-encoding cDNAs (DHFR) containing a Ser31 or a Ser34 mutation, indicating that these human variants can act as dominant selectable markers. Expand
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Analysis of the Pro-3 MtxRIII DHFR cDNA demonstrated a C----T base transition at nucleotide 67 that results in the substitution of phenylalanine for leucine at residue 22 and the loss of a BsaI site, which results in a decreased binding of methotrexate to the altered enzyme. Expand
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