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Phenotypic correction of Fanconi anemia group C knockout mice
1UNC Gene Therapy Center and the Department of Medicine, University of North Carolina, Chapel Hill, NC 27599, USA.
Abstract:
Fanconi anemia (FA) is a genetic disorder characterized by bone marrow failure, congenital anomalies, and a predisposition to malignancy. FA cells demonstrate hypersensitivity to DNA cross-linking agents, such as mitomycin C (MMC). Mice with a targeted disruption of the FANCC gene (fancc -/- nullizygous mice) exhibit many of the characteristic features of FA and provide a valuable tool for testing novel therapeutic strategies. We have exploited the inherent hypersensitivity of fancc -/- hematopoietic cells to assay for phenotypic correction following transfer of the FANCC complementary DNA (cDNA) into bone marrow cells. Murine fancc -/- bone marrow cells were transduced with the use of retrovirus carrying the human fancc cDNA and injected into lethally irradiated recipients. Mitomycin C (MMC) dosing, known to induce pancytopenia, was used to challenge the transplanted animals. Phenotypic correction was determined by assessment of peripheral blood counts. Mice that received cells transduced with virus carrying the wild-type gene maintained normal blood counts following MMC administration. All nullizygous control animals receiving MMC exhibited pancytopenia shortly before death. Clonogenic assay and polymerase chain reaction analysis confirmed gene transfer of progenitor cells. These results indicate that selective pressure promotes in vivo enrichment of fancc-transduced hematopoietic stem/progenitor cells. In addition, MMC resistance coupled with detection of the transgene in secondary recipients suggests transduction and phenotypic correction of long-term repopulating stem cells. (Blood. 2000;95:700-704)
Insights
Gene therapy corrected Fanconi anemia (FA) in mice. Transferring the FANCC gene into bone marrow cells restored blood counts after DNA damage, showing potential for treating this genetic disorder.
Area of Science:
- Hematology
- Genetics
- Molecular Biology
- Gene Therapy
Background:
- Fanconi anemia (FA) is a genetic disorder causing bone marrow failure, congenital anomalies, and cancer predisposition.
- FA cells are hypersensitive to DNA cross-linking agents like mitomycin C (MMC).
- FANCC gene-deficient mice (fancc-/-) serve as a model for FA research and therapeutic strategy testing.
Purpose of the Study:
- To assess phenotypic correction of Fanconi anemia in mouse models.
- To evaluate the efficacy of FANCC complementary DNA (cDNA) gene transfer into hematopoietic stem/progenitor cells.
Main Methods:
- Murine fancc-/- bone marrow cells were transduced with a retrovirus carrying human FANCC cDNA.
- Transduced cells were transplanted into lethally irradiated recipients.
- Mice were challenged with mitomycin C (MMC) to assess phenotypic correction via peripheral blood counts.
Main Results:
- Mice receiving FANCC-transduced cells maintained normal blood counts after MMC exposure.
- Control nullizygous mice receiving MMC exhibited severe pancytopenia.
- Gene transfer and phenotypic correction of long-term repopulating stem cells were confirmed.
Conclusions:
- Selective pressure (MMC) enriches fancc-transduced hematopoietic stem/progenitor cells in vivo.
- Successful gene transfer and phenotypic correction of FA were demonstrated in a murine model.
- This study supports gene therapy as a viable strategy for Fanconi anemia treatment.