title: Phenotype correction of Fanconi anemia group A hematopoietic stem cells using lentiviral vector.
abstract: Fanconi anemia (FA) is an autosomal recessive disease characterized by progressive bone marrow failure due to defective stem cell function. FA patients' cells are hypersensitive to DNA cross-linking agents such as mitomycin C (MMC), exposure to which results in cytogenetic aberrations and cell death. To date Moloney murine leukemia virus vectors have been used in clinical gene therapy. Recently, third-generation lentiviral vectors based on the HIV-1 genome have been developed for efficient gene transfer to hematopoietic stem cells. We generated a self-inactivating lentiviral vector expressing the FA group A cDNA driven by the murine stem cell virus U3 LTR promoter and used the vector to transduce side-population (SP) cells isolated from bone marrow of Fanconi anemia group A (Fanca) knockout mice. One thousand transduced SP cells reconstituted the bone marrow of sublethally irradiated Fanca recipient mice. Phenotype correction was demonstrated by stable hematopoiesis following MMC challenge. Using real-time PCR, one proviral vector DNA copy per cell was detected in all lineage-committed cells in the peripheral blood of both primary and secondary recipients. Our results suggest that the lentiviral vector transduces stem cells capable of self-renewal and long-term hematopoiesis in vivo and is potentially useful for clinical gene therapy of FA hematopoietic cells.
authors.affiliation: Department of Medicine, Mount Sinai School of Medicine, New York, New York 10029, USA.
authors.forename: Kaoru
authors.initials: K
authors.lastname: Yamada
authors.forename: Ali
authors.initials: A
authors.lastname: Ramezani
authors.forename: Robert G
authors.initials: RG
authors.lastname: Hawley
authors.forename: Wolfram
authors.initials: W
authors.lastname: Ebell
authors.forename: Fre
authors.initials: F
authors.lastname: Arwert
authors.forename: Larry W
authors.initials: LW
authors.lastname: Arnold
authors.forename: Christopher E
authors.initials: CE
authors.lastname: Walsh
mesh_terms: D000818:Animals; D016026:Bone Marrow Transplantation; D004268:DNA-Binding Proteins; D005199:Fanconi Anemia / Q000188:drug therapy*; D052217:Fanconi Anemia Complementation Group A Protein; D015316:Genetic Therapy; D005822:Genetic Vectors; D006412:Hematopoietic Stem Cells / Q000378:metabolism; D016086:Lentivirus; D051379:Mice; D018345:Mice, Knockout; D010641:Phenotype; D011506:Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D004268:DNA-Binding Proteins; C120277:Fanca protein, mouse; D052217:Fanconi Anemia Complementation Group A Protein; D011506:Proteins