title: Foamy viral vector integration sites in SCID-repopulating cells after MGMTP140K-mediated in vivo selection.
abstract: Foamy virus (FV) vectors are promising for hematopoietic stem cell (HSC) gene therapy but preclinical data on the clonal composition of FV vector-transduced human repopulating cells is needed. Human CD34(+) human cord blood cells were transduced with an FV vector encoding a methylguanine methyltransferase (MGMT)P140K transgene, transplanted into immunodeficient NOD/SCID IL2Rγ(null) mice, and selected in vivo for gene-modified cells. The retroviral insertion site profile of repopulating clones was examined using modified genomic sequencing PCR. We observed polyclonal repopulation with no evidence of clonal dominance even with the use of a strong internal spleen focus forming virus promoter known to be genotoxic. Our data supports the use of FV vectors with MGMTP140K for HSC gene therapy but also suggests additional safety features should be developed and evaluated.
authors: Department of Pharmaceutical Sciences, Washington State University, Spokane, WA, USA.
authors: M E
authors: ME
authors: Olszko
authors: Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA.
authors: J E
authors: JE
authors: Adair
authors: Department of Pharmaceutical Sciences, Washington State University, Spokane, WA, USA.
authors: I
authors: I
authors: Linde
authors: Department of Pharmaceutical Sciences, Washington State University, Spokane, WA, USA.
authors: D T
authors: DT
authors: Rae
authors: Department of Pharmaceutical Sciences, Washington State University, Spokane, WA, USA.
authors: P
authors: P
authors: Trobridge
authors: Department of Pharmaceutical Sciences, Washington State University, Spokane, WA, USA.
authors: J D
authors: JD
authors: Hocum
authors: Center for Immunity and Immunotherapies, Seattle Children's Research Institute, Seattle, WA, USA.
authors: D J
authors: DJ
authors: Rawlings
authors: Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA.
authors: H-P
authors: HP
authors: Kiem
authors: 1] Department of Pharmaceutical Sciences, Washington State University, Spokane, WA, USA [2] School of Molecular Biosciences, Washington State University, Pullman, Washington, USA.
authors: G D
authors: GD
authors: Trobridge
mesh_terms: D000818:Animals; D005820:Genetic Testing / Q000379:methods; D005822:Genetic Vectors; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells; D006801:Humans; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D016511:Severe Combined Immunodeficiency / Q000235:genetics*; D016092:Spumavirus / Q000235:genetics*; D019172:Transplantation Conditioning; D016662:Virus Integration / Q000235:genetics*