GLUCONOYLATED AND GLYCOSYLATED POLYLYSINES AS VECTORS FOR GENE-TRANSFER INTO CYSTIC-FIBROSIS AIRWAY EPITHELIAL-CELLS

To provide an alternative to viral vectors for the transfer of genes i nto airway epithelial cells in cystic fibrosis (CF), a novel set of su bstituted polylysines were employed, Polylysine was partially neutrali zed by blocking a number of positively charged residues with gluconoyl groups, In addition, polylysine was substituted with sugar residues o n a specified number of amino groups, Using the gluconoylated polylysi ne as vector, the pCMVLuc plasmid gave high expression of the reporter gene luciferase in immortalized CF/T43 cells. The luciferase activity was 75-fold greater in the presence of 100 mu M chloroquine, Lucifera se gene expression persisted at high levels for up to at least 120 hr following transfection, Glycosylated polylysines/pCMVLuc complexes wer e compared to the gluconoylated polylysine/pCMVLuc complex and beta-Ga l-, alpha-Glc-, and Lac-substituted polylysines gave 320%, 300%, and 2 90%, respectively, higher expression of the reporter gene luciferase, Luciferase expression ranged from 35 to 2 ng of luciferase per milligr am of cell protein in the order: beta-Gal = alpha-Glc = Lac > alpha-Ga l = Rha = Man > beta-GalNAc > alpha-GalNAc = alpha-Fuc, suggesting tha t the transfection efficiency is sugar dependent. Most importantly, in primary cultures of both CF and non-CF airway epithelial cells grown from tracheal tissue explants, lactosylated polylysine gave uniformly high expression of luciferase. The glycosylated polylysines provide an attractive nonviral approach for the transfer of genes into airway ep ithelial cells.