BACKGROUND: The development of methods for specific delivery of genes into target tissues is an important issue for the further progress of gene therapy. Biological and physical targeting techniques may be combined to redirect gene therapy vectors to specific cells and enhance gene transfer. METHODS: The polymer poly(2-(dimethylamino)ethyl methacrylate) (pDMAEMA) was conjugated with avidin or poly(ethylene glycol) (PEG) and complexed with adenovirus serotype 5 (Ad5). Targeting of polymer-coated Ad5 to the epidermal growth factor receptor (EGFR) was accomplished by the binding of biotin-EGF to pDMAEMA-avidin. A photochemical treatment procedure using photosensitizer and light was applied to increase transduction with EGFR-targeted viral complexes. RESULTS: pDMAEMA-avidin efficiently enhanced transduction through unspecific viral uptake into cells, while pDMAEMA-PEG provided charge shielding of the complexes and increased the specificity to EGFR when biotin-EGF ligands were used. Transduction of PEG-containing, EGFR-targeted viral complexes was inhibited by 66% in coxsackie and adenovirus receptor (CAR)-deficient RD cells and by 47% in CAR-expressing DU 145 cells in receptor antibody experiments. The photochemical treatment had a substantial effect on transduction, enhancing the percentage of reporter gene positive cells from 20% to 75% of the total viable RD cell population and from 10% to 70% in DU 145 cells. CONCLUSION: Photochemical treatment of cells infected with targeted viral vectors exhibiting a neutral surface charge is a potent method for enhancing transgene expression.
BACKGROUND: The development of methods for specific delivery of genes into target tissues is an important issue for the further progress of gene therapy. Biological and physical targeting techniques may be combined to redirect gene therapy vectors to specific cells and enhance gene transfer. METHODS: The polymerpoly(2-(dimethylamino)ethyl methacrylate) (pDMAEMA) was conjugated with avidin or poly(ethylene glycol) (PEG) and complexed with adenovirus serotype 5 (Ad5). Targeting of polymer-coated Ad5 to the epidermal growth factor receptor (EGFR) was accomplished by the binding of biotin-EGF to pDMAEMA-avidin. A photochemical treatment procedure using photosensitizer and light was applied to increase transduction with EGFR-targeted viral complexes. RESULTS:pDMAEMA-avidin efficiently enhanced transduction through unspecific viral uptake into cells, while pDMAEMA-PEG provided charge shielding of the complexes and increased the specificity to EGFR when biotin-EGF ligands were used. Transduction of PEG-containing, EGFR-targeted viral complexes was inhibited by 66% in coxsackie and adenovirus receptor (CAR)-deficient RD cells and by 47% in CAR-expressing DU 145 cells in receptor antibody experiments. The photochemical treatment had a substantial effect on transduction, enhancing the percentage of reporter gene positive cells from 20% to 75% of the total viable RD cell population and from 10% to 70% in DU 145 cells. CONCLUSION: Photochemical treatment of cells infected with targeted viral vectors exhibiting a neutral surface charge is a potent method for enhancing transgene expression.
Authors: Maximilian Hörner; Carolina Jerez-Longres; Anna Hudek; Sebastian Hook; O Sascha Yousefi; Wolfgang W A Schamel; Cindy Hörner; Matias D Zurbriggen; Haifeng Ye; Hanna J Wagner; Wilfried Weber Journal: Sci Adv Date: 2021-06-16 Impact factor: 14.136