Gene delivery from the E3 region of replicating human adenovirus: evaluation of the ADP region.
Genetically modified replication-selective human adenoviruses are currently undergoing testing in the clinical setting as anticancer agents. Coupling the lytic function of these viruses with virus-mediated transgene delivery represents a powerful extension of this treatment. We have designed a unique system for gene delivery from the replicating virus. It takes advantage of the endogenous gene expression control sequences (promoter, splicing, polyadenylation signals) to efficiently and predictably deliver transgenes from the non-essential E3 transcription unit while still maintaining the expression of the remaining E3 genes in the multi-gene transcription unit. In this article, we engineered restriction enzyme sites into the virus genome selectively to delete the ADP gene and replace it with the therapeutic transgenes CD and TNFalpha. We demonstrate that: (1) transgene expression from this region mirrors the substituted ADP gene; (2) the loss of ADP in these viruses results in infected cells with extended viability and protein synthesis when compared with a wild-type Ad5 infected cell; and (3) expression of surrounding E3 genes can be maintained in such a system. The potential advantages of delivering transgenes from the ADP region of the replicating adenovirus are discussed.
Hawkins
L K
LK
Onyx Pharmaceuticals, 3031 Research Drive, Richmond, CA 94806, USA.
Hermiston
T W
TW
D000244:Adenosine Diphosphate / Q000235:genetics*; D017503:Adenovirus E3 Proteins / Q000235:genetics*; D000260:Adenoviruses, Human; D043525:Cytosine Deaminase; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D017353:Gene Deletion; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage*; D006801:Humans; D016254:Mutagenesis, Insertional / Q000379:methods*; D009700:Nucleoside Deaminases / Q000235:genetics; D014162:Transfection / Q000379:methods*; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis / Q000235:genetics
D017503:Adenovirus E3 Proteins; D014409:Tumor Necrosis Factor-alpha; D000244:Adenosine Diphosphate; D009700:Nucleoside Deaminases; D043525:Cytosine Deaminase