Human cytidine deaminase as an ex vivo drug selectable marker in gene-modified primary bone marrow stromal cells

Eliopoulos, Nicoletta; Al-Khaldi, Abdulaziz; Beauséjour, C.; Rl, Momparler; Momparler, LF; Galipeau, Jacques

doi:10.1038/sj.gt.3301675

Cited by 33 publications

(24 citation statements)

References 49 publications

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“…Alternative human biocompatible selectable markers would be of use in this setting. 80 In conclusion, our data validate the feasibility of utilizing gene-modified autologous bone marrow stroma as a vehicle for sustained systemic production of recombinant therapeutic proteins in immunocompetent recipients, without the major drawback of myeloablation. Furthermore, when delivered subcutaneously in a matrix, MSCs will spontaneously generate a neovascularized organoid.…”

Section: Marrow Stroma Implant For Erythropoietin Delivery In Normal mentioning

confidence: 62%

A neovascularized organoid derived from retrovirally engineered bone marrow stroma leads to prolonged in vivo systemic delivery of erythropoietin in nonmyeloablated, immunocompetent mice

et al. 2003

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Marrow stromal cells (MSCs) are postnatal progenitor cells that can be easily cultured ex vivo to large amounts. This feature is attractive for cell therapy applications where genetically engineered MSCs could serve as an autologous cellular vehicle for the delivery of therapeutic proteins. The usefulness of MSCs in transgenic cell therapy will rely upon their potential to engraft in nonmyeloablated, immunocompetent recipients. Further, the ability to deliver MSCs subcutaneously -as opposed to intravenous or intraperitoneal infusions -would enhance safety by providing an easily accessible, and retrievable, artificial subcutaneous implant in a clinical setting. To test this hypothesis, MSCs were retrovirally engineered to secrete mouse erythropoietin (Epo) and their effect was ascertained in nonmyeloablated syngeneic mice. Epo-secreting MSCs when administered as 'free' cells by subcutaneous or intraperitoneal injection, at the same cell dose, led to a significant -yet temporaryhematocrit increase to over 70% for 55713 days. In contrast, in mice implanted subcutaneously with Matrigeltembedded MSCs, the hematocrit persisted at levels 480% for over 110 days in four of six mice (Po0.05 logrank). Moreover, Epo-secreting MSCs mixed in Matrigel elicited and directly participated in blood vessel formation de novo reflecting their mesenchymal plasticity. MSCs embedded in human-compatible bovine collagen matrix also led to a hematocrit 470% for 7578.9 days. In conclusion, matrixembedded MSCs will spontaneously form a neovascularized organoid that supports the release of a soluble plasma protein directly into the bloodstream for a sustained pharmacological effect in nonmyeloablated recipients.

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Section: Marrow Stroma Implant For Erythropoietin Delivery In Normal mentioning

confidence: 62%

A neovascularized organoid derived from retrovirally engineered bone marrow stroma leads to prolonged in vivo systemic delivery of erythropoietin in nonmyeloablated, immunocompetent mice

et al. 2003

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“…10 Retrovectors encoding green fluorescent protein and the human drug resistance gene cytidine deaminase 11 as well as LacZ 12 were utilized to generate a polyclonal population of gene-marked MSCs. Following LacZ transduction, 70-95% of all gene-modified MSCs expressed detectable b-galactosidase activity as assessed by X-gal staining.…”

Section: Generation Of Lacz Gene-modified Mscsmentioning

confidence: 99%

Postnatal bone marrow stromal cells elicit a potent VEGF-dependent neoangiogenic response in vivo

et al. 2003

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“…[7][8][9] In addition, we and others have shown increased cytarabine resistance after retrovirally mediated transduction of primary murine hematopoietic cells. [10][11][12] More recently, increased gemcitabine and 5-aza-2 0 -deoxycytidine resistance after CDD gene transfer into murine cells 13,14 and transfer of the CDD gene into primary murine bone marrow stromal cells, followed by in vitro enrichment of these cells by cytarabine application, have been described. 14 Successful treatment of lymphoma with a combination of methotrexate and cytarabine in NOD/SCID mice transplanted with drug-resistant bone marrow could also be demonstrated.…”

Section: Introductionmentioning

confidence: 99%

“…[10][11][12] More recently, increased gemcitabine and 5-aza-2 0 -deoxycytidine resistance after CDD gene transfer into murine cells 13,14 and transfer of the CDD gene into primary murine bone marrow stromal cells, followed by in vitro enrichment of these cells by cytarabine application, have been described. 14 Successful treatment of lymphoma with a combination of methotrexate and cytarabine in NOD/SCID mice transplanted with drug-resistant bone marrow could also be demonstrated. 15 Thus far, data for the successful transduction of primary human hematopoietic cells with CDD are missing although human CD34 þ cells have low CDD enzymatic activity, 8 rendering them an excellent target cell population for this approach.…”

Section: Introductionmentioning

confidence: 99%

Resistance to cytarabine and gemcitabine and in vitro selection of transduced cells after retroviral expression of cytidine deaminase in human hematopoietic progenitor cells

et al. 2005

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Overexpression of the detoxifying enzyme cytidine deaminase (CDD) renders normal and leukemic hematopoietic cells resistant to cytarabine (1-b-D-arabinofuranosylcytosine), and studies on murine cells have suggested transgenic CDD overexpression as a way to reduce the substantial myelotoxicity induced by the deoxycytidine analogs cytarabine and gemcitabine (2 0 ,2 0 -difluorodeoxycytidine). We now have investigated CDD (over-)expression in the human hematopoietic system. Retroviral gene transfer significantly increased the resistance of CDD-transduced cord blood and peripheral bloodderived progenitor cells for doses ranging from 20 to 100 nM cytarabine and 8-10 nM gemcitabine. Protection was observed for progenitors of erythroid as well as myeloid differentiation, though the degree of protection varied for individual drugs. In addition, significant selection of CDD-transduced cells was obtained after a 4-day culture in 30-100 nM cytarabine. Thus, our data demonstrate that overexpression of CDD cDNA results in significant protection of human progenitors from cytarabineas well as gemcitabine-induced toxicity, and allows in vitro selection of transduced cells. This strongly argues for a potential therapeutic role of CDD gene transfer in conjunction with dose-intensive cytarabine-or gemcitabine-containing chemotherapy regimen.

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Human cytidine deaminase as an ex vivo drug selectable marker in gene-modified primary bone marrow stromal cells

Cited by 33 publications

References 49 publications

A neovascularized organoid derived from retrovirally engineered bone marrow stroma leads to prolonged in vivo systemic delivery of erythropoietin in nonmyeloablated, immunocompetent mice

A neovascularized organoid derived from retrovirally engineered bone marrow stroma leads to prolonged in vivo systemic delivery of erythropoietin in nonmyeloablated, immunocompetent mice

Postnatal bone marrow stromal cells elicit a potent VEGF-dependent neoangiogenic response in vivo

Resistance to cytarabine and gemcitabine and in vitro selection of transduced cells after retroviral expression of cytidine deaminase in human hematopoietic progenitor cells

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