Dystrophin Expression in Muscle Following Gene Transfer with a Fully Deleted ("Gutted") Adenovirus Is Markedly Improved by Trans-Acting Adenoviral Gene Products
- 20 September 2001
- journal article
- research article
- Published by Mary Ann Liebert Inc in Human Gene Therapy
- Vol. 12 (14), 1741-1755
- https://doi.org/10.1089/104303401750476249
Abstract
Helper-dependent adenoviruses (HDAd) are Ad vectors lacking all or most viral genes. They hold great promise for gene therapy of diseases such as Duchenne muscular dystrophy (DMD), because they are less immunogenic than E1/E3-deleted Ad (first-generation Ad or FGAd) and can carry the full-length (Fl) dystrophin (dys) cDNA (12 kb). We have compared the transgene expression of a HDAd (HDAdCMVDysFl) and a FGAd (FGAdCMV-dys) in cell culture (HeLa, C2C12 myotubes) and in the muscle of mdx mice (the mouse model for DMD). Both vectors encoded dystrophin regulated by the same cytomegalovirus (CMV) promoter. We demonstrate that the amount of dystrophin expressed was significantly higher after gene transfer with FGAdCMV-dys compared to HDAdCMVDysFl both in vitro and in vivo. However, gene transfer with HDAdCMVDysFl in the presence of a FGAd resulted in a significant increase of dystrophin expression indicating that gene products synthesized by the FGAd increase, in trans, the amount of dystrophin produced. This enhancement occurred in cell culture and after gene transfer in the muscle of mdx mice and dystrophic golden retriever (GRMD) dogs, another animal model for DMD. The E4 region of Ad is required for the enhancement, because no increase of dystrophin expression from HDAdCMVDysFl was observed in the presence of an E1/E4-deleted Ad in vitro and in vivo. The characterization of these enhancing gene products followed by their inclusion into an HDAd may be required to produce sufficient dystrophin to mitigate the pathology of DMD by HDAd-mediated gene transfer.Keywords
This publication has 74 references indexed in Scilit:
- Differential influence of the E4 adenoviral genes on viral and cellular promotersThe Journal of Gene Medicine, 2000
- DNA Replication of First-Generation Adenovirus Vectors in Tumor CellsHuman Gene Therapy, 2000
- Prolonged Transgene Expression Mediated by a Helper-Dependent Adenoviral Vector (hdAd) in the Central Nervous SystemMolecular Therapy, 2000
- Increased Duration of Transgene Expression in the Lung with Plasmid DNA Vectors Harboring Adenovirus E4 Open Reading Frame 3Human Gene Therapy, 1999
- Persistence of an [E1-, Polymerase-] Adenovirus Vector Despite Transduction of a Neoantigen into Immune-Competent MiceHuman Gene Therapy, 1999
- Efficient Utrophin Expression Following Adenovirus Gene Transfer in Dystrophic MuscleBiochemical and Biophysical Research Communications, 1998
- Adenovirus-mediated gene transfer: influence of transgene, mouse strain and type of immune response on persistence of transgene expressionGene Therapy, 1997
- Persistent transgene expression in mouse liver following in vivo gene transfer with a ΔE1/ΔE4 adenovirus vectorGene Therapy, 1997
- Inactivation of E2a in recombinant adenoviruses improves the prospect for gene therapy in cystic fibrosisNature Genetics, 1994
- Selective extraction of polyoma DNA from infected mouse cell culturesJournal of Molecular Biology, 1967