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 beta(2)-adrenergic receptor (beta(2)AR) cDNA. Human beta(2)AR message and protein expression were noted 2-4 h postinfection without evidence of pseudotransduction. beta(2)AR function (cAMP production) was increased at 6 h postinfection. To determine when beta(2)AR gene transfer affects downstream catecholamine-sensitive pathways, we measured lung Na,K-ATPase expression and alveolar fluid clearance (AFC). beta(2)AR 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.

Increased Expression of Integrin-Linked Kinase Attenuates Left Ventricular Remodeling and Improves Cardiac Function After Myocardial Infarction

“…3 Although we detected ILK expression from 3 days after infarction in our study, it is known that recombinant adenoviruses can produce physiologically significant levels of transgene as early as 2 to 4 hours after infection, 15 rendering it possible that the effect of adeno-ILK in this system commenced earlier than 3 days and within the early phase of remodeling. Our results demonstrate that expression of ILK itself and hrGFP disappeared by 35 days (5 weeks), whereas the effect of ILK treatment persisted for at least 7 weeks, as judged by hemodynamic alterations as well as structural and morphological changes in the heart.…”

Lung-specific loss of the laminin α3 subunit confers resistance to mechanical injury

“…It remains possible, however, that knockdown of a3 laminin in the airways, or other cell types within the lung, might have contributed to the observed phenotype. In addition, although wild-type mice infected with an adenovirus develop inflammation that is maximal 2 days after the infection and is undetectable after 10 days, we cannot exclude the possibility that the inflammation and collagen deposition we observed in the lungs upon a3 laminin knockdown at 60 days after infection results from impaired resolution of virally induced lung injury (Dumasius et al, 2003).…”

“…Adenoviruses preferentially infect cells expressing the Cocksakie A receptor (CAR), a junctional adhesion molecule (JAM) localized to tight junctions in the airway and alveolar epithelium (Cohen et al, 2001). Following infection with adenoviruses, expression of the transgene is maximal after 48 hours and is not detectable after 10 days (Dumasius et al, 2003). Using b-galactosidase Cre reporter mice, we have previously shown that administration of adenoviral Cre at this dose results in widespread recombination in the airway and alveolar epithelium Weinberg et al, 2010).…”

Intraplacental Gene Therapy with Ad-IGF-1 Corrects Naturally Occurring Rabbit Model of Intrauterine Growth Restriction

“…The type of disease to be treated needs to be defined before decisions are made as to which vector type should be applied. 45 Adenoviral vector is appealing for intraplacental gene therapy because it is replication deficient, it demonstrates early onset of transgenic protein expression (within 24 hr), wide transduction when driven by a CMV promoter, 46,47 and short-term transgenic protein expression (typically ranging from 10 to 14 days). 48 Although early versions of adenoviruses showed toxic side effects and strong immune responses, newer second-and third-generation vectors with many of the viral genes deleted have demonstrated significant improvements.…”