In vivo timing of onset of transgene expression following adenoviral-mediated gene transfer

Abstract

Recombinant adenoviruses are efficient gene transfer vehicles that could be used for treatment of acute diseases. However, the time required for adenoviruses to produce physiologically relevant levels of transgene in vivo is unknown. To address this question rat lungs were infected with an E1a −/E3a − adenovirus that contains an hCMV-driven human β 2-adrenergic receptor (β 2AR) cDNA. Human β 2AR message and protein expression were noted 2–4 h postinfection without evidence of pseudotransduction. β 2AR function (cAMP production) was increased at 6 h postinfection. To determine when β 2AR gene transfer affects downstream catecholamine-sensitive pathways, we measured lung Na,K-ATPase expression and alveolar fluid clearance (AFC). β 2AR gene transfer increased Na,K-ATPase number by 80% at 6 h, and AFC by 20% at 8 h postinfection. These data indicate that recombinant adenoviruses can produce physiologically significant levels of transgene within hours of infection and that they may be suitable for gene therapies for acute, rapidly progressive diseases.