Expression of retroviral vectors containing the human multidrug resistance 1 cDNA in hematopoietic cells of transplanted mice

Sorrentino, Brian P.; McDonagh, Kevin T.; Woods, Diane R.; Orlic, Donald

doi:10.1182/blood.v86.2.491.bloodjournal862491

Cited by 99 publications

(25 citation statements)

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“…As a consequence of these studies, expression of ABC transporters to protect against multiple drugs has been actively pursued. Methods for optimizing gene transfer and expression of a splice-corrected MDR1 vector [42] led to high-level expression of the human P-gp in murine stem cells. It was first reported by Bunting et al that Harvey murine sarcoma virus (Ha)MDR1-transduced BM cells showed a significant engraftment advantage relative to those BM cells transduced with a dihydrofolate reductase (DHFR) mutant containing a leucine to tyrosine mutation at codon 22 (HaDHFR L22Y ) [43].…”

Section: Functional Characterization Of Abc Transporters By Overexprementioning

confidence: 99%

ABC Transporters as Phenotypic Markers and Functional Regulators of Stem Cells

2002

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Section: Functional Characterization Of Abc Transporters By Overexprementioning

confidence: 99%

ABC Transporters as Phenotypic Markers and Functional Regulators of Stem Cells

2002

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“…ABCB1 is also expressed in primitive stem cells, including human CD34 stem cells [4] and c-kit+ cells [5], which can be identified by their ability to transport fluorescent dyes that are ABCB1 substrates, such as rhodamine 123 (Rh123) [6][7][8] and Hoechst 33342 [9][10][11]. One way to purify these stem cells is based on Hoechst dye staining of bone marrow cells to identify a minor fraction of Ôside-populationÕ (SP) cells; this population shows a high repopulating activity and ABCG2 expression [12,13].…”

Section: Introductionmentioning

confidence: 99%

Characterization of ABC transporter ABCB1 expressed in human neural stem/progenitor cells

Islam¹,

Kanemura²,

Tajria³

et al. 2005

FEBS Letters

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We investigated the localization and functional expression of the ABC transporter ABCB1 in human fetal neural stem/progenitor cells (hNSPCs). RT-PCR analysis revealed ABCB1 gene expression in hNSPCs. We found a single band in immunoblotted hNSPCs lysates probed with ABCB1 antibody, and detected ABCB1 at the hNSPCs cell membrane by immunocytochemistry and subcellular fractionation. ABCB1 inhibitors and substrate, and ATP-depleting agents enhanced hNSPCsÕ rhodamine 123 accumulation, and hNSPCs microsomes had vanadate-sensitive ATPase activity. ABCB1 and nestin expression decreased during hNSPCs differentiation, while the astroglial marker GFAP increased. ABCB1 may maintain hNSPCs in an undifferentiated state and could be a neural stem/progenitor marker.

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“…It is also not certain which vector will provide optimal expression of DHFR in early myeloid cells. We chose the Harvey vector based on data showing significant mRNA expression in primitive murine myeloid cells [28]. The recent description of retroviral vectors isolated for high level expression in stem cells [29-311 may offer even greater levels of stem cell expression and selection.…”

Section: Discussionmentioning

confidence: 99%

Retroviral vectors containing a variant dihydrofolate reductase gene for drug protection and in vivo selection of hematopoietic cells

et al. 2009

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Transfer of drug resistance genes to hematopoietic cells is being studied as a means to protect against the myelosuppression associated with cancer chemotherapy and as a strategy for the in vivo selection and amplification of genetically modified cells. The goal of this study was to test if retroviral-mediated gene transfer of a dihydrofolate reductase (DHFR) variant (L22Y) could be used for in vivo selection of transduced myeloid cells and to determine what proportion of transduced cells was required for protection from myelosuppression. Based on previous work suggesting that selection with antifolates may also require inhibition of nucleoside transport mechanisms, mice transplanted with DHFR-transduced bone marrow cells were treated with trimetrexate and the nucleoside transport inhibitor prodrug nitrobenzylmercaptopurine riboside phosphate. In vivo selection of transduced myeloid progenitors was seen in the bone marrow and in circulating mature peripheral blood cells following drug treatment. These results show that the novel combination of the L22Y-DHFR cDNA, trimetrexate and nitrobenzylmercaptopurine riboside phosphate can be used to select for transduced myeloid cells, and that this approach warrants further study in large animal models. A bicistronic vector containing a human CD24 reporter gene was used to determine the number of modified cells needed for chemoprotection. Partial protection from neutropenia was seen when greater than 10% of myeloid cells expressed the vector, and high levels of protection were obtained when the proportion exceeded 30%. These results suggest that gene transfer may be useful for myeloprotection in certain pediatric cancers, but that more efficient gene transfer will be required to apply this approach to adult cancer patients.

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Expression of retroviral vectors containing the human multidrug resistance 1 cDNA in hematopoietic cells of transplanted mice

Cited by 99 publications

References 0 publications

ABC Transporters as Phenotypic Markers and Functional Regulators of Stem Cells

ABC Transporters as Phenotypic Markers and Functional Regulators of Stem Cells

Characterization of ABC transporter ABCB1 expressed in human neural stem/progenitor cells

Retroviral vectors containing a variant dihydrofolate reductase gene for drug protection and in vivo selection of hematopoietic cells

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