Approaches to Enhance Expression After Adenovirus-Mediated Gene Transfer to the Carotid Artery

The goal of this study was to enhance transgene expression after adenoviral-mediated gene transfer to the carotid artery. We used an adenoviral vector with a transgene that expresses β-galactosidase, driven by the human cytomegalovirus (CMV) promoter/enhancer. The CMV promoter drives constitutive expression, and response elements within the enhancer allow inducible expression through binding of active transcription factors, such as cAMP response element binding protein (CREB) and nuclear factor kappa B (NFkB). Rings of rabbit carotid artery were incubated ex vivo with a replication-deficient adenovirus that expresses β-galactosidase (AdCMV-βgal). Virus was removed from the medium, and forskolin or phorbol-12-myristate-13-acetate (PMA), which can induce activation of CREB or NFKB, respectively, were added to the medium. Pyrrolidine dithiocarbamate (PDTC) was used to inhibit activation of NFkB. Following incubation for 24 hours, β-galactosidase activity was assessed by chemiluminescent reporter assay. Forsk...