Ex vivo culture of Fancc-/- stem/progenitor cells predisposes cells to undergo apoptosis, and surviving stem/progenitor cells display cytogenetic abnormalities and an increased risk of malignancy

Li, Xiaxin; Beau, Michelle M. Le; Ciccone, Samantha; Yang, Feng Chun; Freie, Brian; Chen, Shi; Yuan, Jin; Hong, Ping; Orazi, Attilio; Haneline, Laura S.; Clapp, D. Wade

doi:10.1182/blood-2004-06-2483

Cited by 62 publications

(87 citation statements)

References 51 publications

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“…Those abnormalities appeared to arise specifically from cells that had undergone prolonged transduction culture, but remained untransduced, and, consequently, genetically uncorrected. 21 Finally, this protocol represents an ideal platform for subsequent in vivo selection to achieve complete phenotype correction and high-level therapeutic chimerism required for some applications. Clearly, studies in large animal models and targeting human CD34-enriched progenitor cell populations are needed to confirm the feasibility of our strategy.…”

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confidence: 99%

Rapid 1-hour transduction of whole bone marrow leads to long-term repopulation of murine recipients with lentivirus-modified hematopoietic stem cells

2005

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Efficient gene transfer to hematopoietic stem cells by Moloney murine leukemia virus-derived retroviral vectors benefits from ex vivo culture and cytokine support. Both also increase the risks of apoptosis and differentiation among cells targeted for transduction. In an effort to maximize the retention of stem cell properties in target cells, we developed a transduction protocol with a focus on minimizing graft manipulation, cytokine stimulation, and ex vivo exposure duration. Based on their wide host range and ability to transduce quiescent cells, human immunodeficiency virus (HIV)-derived lentivirus vectors are ideally suited for this purpose. Our present studies in a murine model show that whole bone marrow cells are readily transduced after a 1-hour vector exposure in the presence of stem cell factor and CH296 fibronectin fragment. Using this rapid transduction protocol, we achieved long-term, multilineage reconstitution of murine recipients with up to 25% GFP-expressing cells in primary and secondary recipients. Our results demonstrate the unique ability of HIV-derived vectors to transduce hematopoietic stem cells in the absence of enrichment, under minimal cytokine stimulation, and following brief exposures.

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confidence: 99%

Rapid 1-hour transduction of whole bone marrow leads to long-term repopulation of murine recipients with lentivirus-modified hematopoietic stem cells

2005

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“…FA-D1, FA-D2, FA-M), AML is not spontaneously generated in the FA mouse models generated so far. Studies conducted in Fancc -/-mice have shown, however, that the ex vivo culture and/or incubation of Fancc -/-BM cells with TNF-α, whose expression is significantly increased in FA patients, induces leukemic clonal evolution after transplantation [109,110], suggesting that leukemia development in FA patients could be at least partially related to the deregulated expression of this cytokine. Fanca -/-as well as Fancc -/-mice, and also mice with a hypomorphic mutation in Brca2/Fancd1 (FA-D1 mice) constitute the FA mouse models more frequently used both to understand the role of FA genes in HSCs functionality, and also to evaluate the preclinical efficacy of new therapies in FA.…”

Section: Lessons From Fanconi Anemia Mouse Modelsmentioning

confidence: 99%

From the Molecular Biology to the Gene Therapy of a DNA Repair Syndrome: Fanconi Anemia

Rı́o¹,

Navarro²,

Bueren³

2011

DNA Repair and Human Health

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“…Competitive repopulation experiments were conducted as previously described in primary and secondary recipients [36,38]. Briefly, LDMC from 1 ml (cohort 1) or 2 ml (cohort 2) of the PB of donor WT, Fanca −/− or Fancc −/− mice treated with G-CSF, AMD3100 or both were transplanted along with 7.5 × 10 5 BoyJ competitor BM LDMC into WT (8-12 week old) lethally irradiated recipients (1100 rads).…”

Section: Competitive Repopulation Assaysmentioning

confidence: 99%

“…The identification of these genes raises the potential of using gene transfer technology to express the functional cDNA in autologous stem cells. The BM hypoplasia commonly observed in FA patients [1] and the reduced repopulating ability of stem cells in mice containing a disruption of the murine homologue of an FA gene [29,30] led to the hypothesis that autologous, geneticallycorrected stem cells would have an engraftment and proliferation advantage.…”

Section: Introductionmentioning

confidence: 99%

AMD3100 synergizes with G-CSF to mobilize repopulating stem cells in Fanconi anemia knockout mice

Pulliam

Hobson

Ciccone

et al. 2008

Experimental Hematology

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Fanconi anemia (FA) is a heterogeneous inherited disorder characterized by a progressive bone marrow (BM) failure and susceptibility to myeloid leukemia. Genetic correction using gene transfer technology is one potential therapy. A major hurdle in applying this technology in FA patients is the inability of granulocyte colony-stimulating factor (G-CSF) to mobilize sufficient numbers of hematopoietic stem (HSC)/progenitor cells (HPC) from the BM to the PB. Whether the low number of CD34 + cells is a result of BM hypoplasia or an inability of G-CSF to adequately mobilize FA HSC/HPC remains incompletely understood. Here we use competitive repopulation of lethally irradiated primary and secondary recipients to show that in two murine models of FA, AMD3100 synergizes with G-CSF resulting in a mobilization of HSC, whereas G-CSF alone fails to mobilize stem cells even in the absence of hypoplasia.

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Ex vivo culture of Fancc-/- stem/progenitor cells predisposes cells to undergo apoptosis, and surviving stem/progenitor cells display cytogenetic abnormalities and an increased risk of malignancy

Cited by 62 publications

References 51 publications

Rapid 1-hour transduction of whole bone marrow leads to long-term repopulation of murine recipients with lentivirus-modified hematopoietic stem cells

Rapid 1-hour transduction of whole bone marrow leads to long-term repopulation of murine recipients with lentivirus-modified hematopoietic stem cells

From the Molecular Biology to the Gene Therapy of a DNA Repair Syndrome: Fanconi Anemia

AMD3100 synergizes with G-CSF to mobilize repopulating stem cells in Fanconi anemia knockout mice

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