Transfer of Drug-Resistance Genes into Hematopoietic Progenitors

Koç, Omer N.

doi:10.1016/b978-012437551-2/50022-7

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“…Our results are similar to reports describing enrichment of hematopoietic progenitors using drug resistance genes (26,27), but this is the first application of this strategy to bone marrow mesenchymal progenitors. Drug resistance gene therapy of hematopoietic progenitors is explored to overcome the low transduction efficiency by selectively expanding small population of transduced stem cells in vivo (28). hMSCs can be transduced at high frequency, but their use in transplantation has been hampered by poor homing and engraftment after i.v.…”

Section: Discussionmentioning

confidence: 99%

G156A MGMT-Transduced Human Mesenchymal Stem Cells Can Be Selectively Enriched byO⁶-Benzylguanine and BCNU

Lee

Gerson²,

Maitra

et al. 2001

Journal of Hematotherapy & Stem Cell Research

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Human bone marrow-derived mesenchymal stem cells (hMSCs) are being investigated for a potential therapeutic role as hematopoietic support cells following chemo-radiotherapy and as vehicles of gene delivery. Although hMSCs can be safely infused into humans and experimental animals, there is limited evidence regarding their engraftment and proliferation in vivo. We developed a drug resistance gene transfer strategy to mark and selectively enrich marked hMSCs using chemotherapy. We have determined that hMSCs are markedly sensitized to 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) in vitro when pretreated with O(6)-benzylguanine (BG) resulting in a more than four-fold decrease in BCNU IC(90). The MFG retroviral vector encoding a bicistronic transcript for green fluorescent protein (GFP) and mutant (G156A)-methylguanine methyltransferase (G156A-MGMT), which encodes O(6)-alkylguanine-DNA alkyltransferase (AGT), conferring, BG plus BCNU resistance, transduced a high percentage of hMSCs. Transduced hMSCs had high expression of GFP and AGT and became significantly resistant to BG and BCNU. Furthermore, the proportion of GFP expressing transduced hMSCs increased from 32 +/- 14% to 70 +/- 14% following BG and BCNU treatment in vitro. Intravenously infused hMSCs were detected in NOD-SCID mice 8 weeks later by PCR analysis but could not be recultured from the bone marrow. GFP-expressing hMSCs inoculated into subcutaneous wounds in nonobese diabetic-severe combined immunodeficient (NOD-SCID) mouse could be recultured at a low frequency, but enriched by BG and BCNU treatment from 0.05 +/- 0.03% to 0.55 +/- 0.4 (p = 0.028, Welch t-test). Our results indicate that hMSCs are sensitive to BG and BCNU, predicting significant toxicity to the hematopoietic microenvironment with this therapy. G156A-MGMT is a powerful selectable gene for a second marker gene in hMSCs. Drug resistance gene transfer into hMSCs may allow in vivo enrichment of hMSCs when MSC homing and engraftment into target tissues is optimized.

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Section: Discussionmentioning

confidence: 99%

G156A MGMT-Transduced Human Mesenchymal Stem Cells Can Be Selectively Enriched byO⁶-Benzylguanine and BCNU

Lee

Gerson²,

Maitra

et al. 2001

Journal of Hematotherapy & Stem Cell Research

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Transfer of Drug-Resistance Genes into Hematopoietic Progenitors

Cited by 1 publication

References 64 publications

G156A MGMT-Transduced Human Mesenchymal Stem Cells Can Be Selectively Enriched byO⁶-Benzylguanine and BCNU

G156A MGMT-Transduced Human Mesenchymal Stem Cells Can Be Selectively Enriched byO⁶-Benzylguanine and BCNU

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Transfer of Drug-Resistance Genes into Hematopoietic Progenitors

Cited by 1 publication

References 64 publications

G156A MGMT-Transduced Human Mesenchymal Stem Cells Can Be Selectively Enriched byO6-Benzylguanine and BCNU

G156A MGMT-Transduced Human Mesenchymal Stem Cells Can Be Selectively Enriched byO6-Benzylguanine and BCNU

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G156A MGMT-Transduced Human Mesenchymal Stem Cells Can Be Selectively Enriched byO⁶-Benzylguanine and BCNU

G156A MGMT-Transduced Human Mesenchymal Stem Cells Can Be Selectively Enriched byO⁶-Benzylguanine and BCNU