title: In vivo retroviral gene transfer by direct intrafemoral injection results in correction of the SCID phenotype in Jak3 knock-out animals.
abstract: Efficient retroviral gene transfer to pluripotential hematopoietic stem cells (PHSCs) requires ex vivo culture in multiple hematopoietic growth factors (HGFs) to promote cell division. While treatment of PHSCs with HGF can render stem cells viable targets for retroviral infection, HGFs can promote differentiation, loss of self-renewal potential, and affect the homing/engraftment capacity of PHSCs. To avoid the negative impacts observed with ex vivo transduction protocols, we developed a murine model for in vivo retroviral infection by direct intrafemoral injection (DII), thus abolishing the need for removal of cells from their native microenvironment and the signals necessary to maintain their unique physiology. Using this approach we have demonstrated in vivo retroviral gene transfer to colony-forming units-c (CFU-c), short-term reconstituting cells, and PHSCs. Moreover, direct intrafemoral injection of Jak3 knock-out mice with retroviral particles encoding the Jak3 gene resulted in reconstitution of normally deficient lymphocyte populations concomitant with improved immune function. In addition, DII can be used to target the delivery of other gene therapy vectors including adenoviral vectors to bone marrow cells in vivo. Taken together, these results demonstrate that in vivo retroviral gene transfer by direct intrafemoral injection may be a viable alternative to current ex vivo gene transfer approaches.
authors.affiliation: Basic Research Program, SAIC-Frederick, MD, USA.
authors.forename: Christine S
authors.initials: CS
authors.lastname: McCauslin
authors.forename: John
authors.initials: J
authors.lastname: Wine
authors.forename: Linzhao
authors.initials: L
authors.lastname: Cheng
authors.forename: Kim D
authors.initials: KD
authors.lastname: Klarmann
authors.forename: Fabio
authors.initials: F
authors.lastname: Candotti
authors.forename: Peter A
authors.initials: PA
authors.lastname: Clausen
authors.forename: Sally E
authors.initials: SE
authors.lastname: Spence
authors.forename: Jonathan R
authors.initials: JR
authors.lastname: Keller
mesh_terms: D000818:Animals; D005269:Femur; D018014:Gene Transfer Techniques; D007167:Immunotherapy; D007273:Injections, Intramuscular; D053616:Janus Kinase 3; D051379:Mice; D018345:Mice, Knockout; D010641:Phenotype; D039904:Pluripotent Stem Cells / Q000378:metabolism / Q000637:transplantation; D011505:Protein-Tyrosine Kinases / Q000008:administration & dosage* / Q000235:genetics / Q000627:therapeutic use; D012190:Retroviridae / Q000235:genetics; D016511:Severe Combined Immunodeficiency / Q000628:therapy*; D033581:Stem Cell Transplantation
chemical_list: D011505:Protein-Tyrosine Kinases; C507935:Jak3 protein, mouse; D053616:Janus Kinase 3