Stable in vivo gene transduction via a novel adenoviral/retroviral chimeric vector

Abstract

Gene therapy to correct defective genes requires efficient gene delivery and long-term gene expression. The available vector systems have not allowed the simultaneous achievement of both goals. We have developed a chimeric viral vector system that incorporates favorable aspects of both adenoviral and retroviral vectors. Adenoviral vectors induce target cells to function as transient retroviral producer cells in vivo. The progeny retroviral vector particles are then able to stably transduce neighboring cells. In this system, the nonintegrative adenoviral vector is rendered functionally integrative via the intermediate generation of a retroviral producer cell. The chimeric vectors may allow realization of the requisite goals for specific gene-therapy applications.

Transgene amplification and persistence after delivery of retroviral vector and packaging functions with E1/E4-deleted adenoviruses

“…23 Finally, two E1-defective adenoviruses have been used successfully to deliver MoMLVbased vectors. 30 Interestingly, this chimeric vector system allowed efficient and stable transgene delivery both in vitro and in vivo.…”

Post-mitotic, differentiated myotubes efficiently produce retroviral vector from hybrid adeno-retrovirus templates

“…In an attempt to circumvent these problems, improve retroviral vector production and examine the role that individual components of retrovirus play during viral assembly, adeno-retroviral hybrid vectors are being developed by a number of groups. [21][22][23][24][25][26] This new vector system offers the advantageous production and transduction properties of adenovirus in conjunction with the proviral integration properties of retrovirus. 26 Production of functional retroviral vector using this hybrid system is a two-step process; target cells are infected with adenoviruses expressing retrovirus structural genes and provirus sequences.…”

“…Indeed vectors constructed using this approach have already been used in two rodent models of cancer and shown to efficiently transduce rapidly dividing cancer cells. 21,23 One aspect of DMD pathology makes it an ideal target for gene therapy by in situ delivery of retroviral vector. Muscle fibres not expressing dystrophin degenerate and are subsequently replaced by proliferating myoblast stem cells during regeneration.…”

“…Adenoviruses expressing retroviral structural genes Gag, Pol and Env driven by the CMV promoter (AdCMVAmpg) and retroviral provirus expressing GFP (AdLNCMVGFP) ( Figure 1b) were kind gifts from David Curiel (Department of Pathology, University of Alabama, Birmingham, AL, USA) and have been described previously. 23 AdGagPol expressing Gag and Pol driven from a CMV promoter has previously been described (Figure 1b). 22 Ad10A1 expressing the amphotropic envelope protein 10A1 from the elongation factor 1 (EF-1) promoter (Figure 1b) was constructed using a similar approach to that adopted for the construction of AdGagPol.…”

“…Cells were induced to fully differentiate into post-mitotic myotubes by replacing C2C12 growth medium with DMEM containing 5.0% 28 22 as has construction of AdLNCMVGFP and AdCMVAmpg. 23 horse serum (HS). Partial differentiation was achieved by incubating confluent cultures of C2C12 cells with DMEM containing 0.5% FCS.…”