Tat conjugation of adenovirus vector broadens tropism and enhances transduction efficiency

Abstract

Aims Adenovirus vectors (Advs) have been very useful for basic research and clinical gene therapy because they propagate to high titers and efficiently transduce cells and tissues regardless of the mitotic status. However, poor transduction of cells that lack the coxsackievirus and adenovirus receptor (CAR), the primary receptor for Advs, has limited Adv application. In this study, we attempted to generate novel Tat–Advs (Advs conjugated with the HIV Tat-derived peptide, a protein-transduction domain (PTD)) to broaden Adv tropism and enhance transduction efficiency. Main methods We constructed Tat–Advs by chemically conjugating Tat peptide to the surface-exposed lysine residues on Advs. We compared the gene transfer activity of Tat–Advs with that of unmodified Advs by measuring the luciferase expression in several types of cell lines. Key findings Tat–Advs showed gene expression 1 to 3 log orders higher than unmodified Advs in CAR-negative adherent cells and blood cells, which are refractory to conventional Advs. The inhibition of Tat–Adv-mediated gene expression by heparin and macropinocytosis inhibitor confirms that binding of Tat–Adv to cellular HSPGs and macropinocytosis are essential for efficient CAR-independent transduction. We also demonstrated that Adv modified with another PTD (R8) had the same high transduction efficiency as Tat–Adv. Significance These data suggest that Tat–Advs are important tools for transducing cells and will be useful as platform vectors for gene therapy.