title: Intraosseous delivery of lentiviral vectors targeting factor VIII expression in platelets corrects murine hemophilia A.
abstract: Intraosseous (IO) infusion of lentiviral vectors (LVs) for in situ gene transfer into bone marrow may avoid specific challenges posed by ex vivo gene delivery, including, in particular, the requirement of preconditioning. We utilized IO delivery of LVs encoding a GFP or factor VIII (FVIII) transgene directed by ubiquitous promoters (a MND or EF-1α-short element; M-GFP-LV, E-F8-LV) or a platelet-specific, glycoprotein-1bα promoter (G-GFP-LV, G-F8-LV). A single IO infusion of M-GFP-LV or G-GFP-LV achieved long-term and efficient GFP expression in Lineage(-)Sca1(+)c-Kit(+) hematopoietic stem cells and platelets, respectively. While E-F8-LV produced initially high-level FVIII expression, robust anti-FVIII immune responses eliminated functional FVIII in circulation. In contrast, IO delivery of G-F8-LV achieved long-term platelet-specific expression of FVIII, resulting in partial correction of hemophilia A. Furthermore, similar clinical benefit with G-F8-LV was achieved in animals with pre-existing anti-FVIII inhibitors. These findings further support platelets as an ideal FVIII delivery vehicle, as FVIII, stored in α-granules, is protected from neutralizing antibodies and, during bleeding, activated platelets locally excrete FVIII to promote clot formation. Overall, a single IO infusion of G-F8-LV was sufficient to correct hemophilia phenotype for long term, indicating that this approach may provide an effective means to permanently treat FVIII deficiency.
authors: Center for Immunity and Immunotherapies, Seattle Children's Research Institute, Seattle, Washington, USA.
authors: Xuefeng
authors: X
authors: Wang
authors: Center for Immunity and Immunotherapies, Seattle Children's Research Institute, Seattle, Washington, USA.
authors: Simon C
authors: SC
authors: Shin
authors: Center for Immunity and Immunotherapies, Seattle Children's Research Institute, Seattle, Washington, USA.
authors: Andy F J
authors: AF
authors: Chiang
authors: Center for Immunity and Immunotherapies, Seattle Children's Research Institute, Seattle, Washington, USA.
authors: Iram
authors: I
authors: Khan
authors: Division of Experimental Hematology, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA.
authors: Dao
authors: D
authors: Pan
authors: 1] Center for Immunity and Immunotherapies, Seattle Children's Research Institute, Seattle, Washington, USA [2] Department of Pediatrics, University of Washington, Seattle, Washington, USA.
authors: David J
authors: DJ
authors: Rawlings
authors: 1] Center for Immunity and Immunotherapies, Seattle Children's Research Institute, Seattle, Washington, USA [2] Department of Pediatrics, University of Washington, Seattle, Washington, USA.
authors: Carol H
authors: CH
authors: Miao
mesh_terms: D000818:Animals; D001792:Blood Platelets / Q000378:metabolism*; D002460:Cell Line; D005169:Factor VIII / Q000235:genetics* / Q000378:metabolism; D005822:Genetic Vectors / Q000008:administration & dosage*; D049452:Green Fluorescent Proteins / Q000235:genetics; D006467:Hemophilia A / Q000097:blood / Q000628:therapy*; D006801:Humans; D017148:Infusions, Intraosseous; D016086:Lentivirus / Q000235:genetics*; D051379:Mice
chemical_list: D049452:Green Fluorescent Proteins; D005169:Factor VIII
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