Efficient transfer of genes into senescent cells by adenovirus vectors via highly expressed alpha v beta 5 integrin

Although various methods for transferring genes into mammalian cells have been established, none have been successful with senescent cells. In this report, we present evidence of the efficient transfer of a gene into human senescent fibroblasts using an adenoviral vector. By employing a recombinant adenovirus vector harboring the beta-galactosidase gene (Ad-CAG beta NR), we observed a good correlation between the proportion of beta-galactosidase positive cells and population doubling of the infected cells. In addition, 1.5- to 6.0-fold greater beta-galactosidase activity was observed in senescent fibroblasts (population doubling [PD] = 58) than in young cells (PD = 15). Western blotting analysis revealed that, compared with young fibroblasts, senescent fibroblasts expressed larger amounts of alpha v beta 5 and alpha v beta 3 integrins which were thought to form part of the adenovirus receptor. These results suggest that higher expression of alpha v beta 5 and alpha v beta 3 integrins in senescent cells renders them more sensitive to adenovirus infection than young cells. Thus, adenovirus vectors may prove to be useful in gene therapy strategies directed against senescence-related disorders.