Gluconoylated and glycosylated polylysines as vectors for gene transfer into cystic fibrosis airway epithelial cells

Wouter J.W. Kollen, Patrick Midoux, Patrick Erbacher, Alex Yip, Annie Claude Roche, Michel Monsigny, Mary Catherine Glick, Thomas F. Scanlin

Research output: Contribution to journalArticle

46 Scopus citations

Abstract

To provide an alternative to viral vectors for the transfer of genes into airway epithelial cells in cystic fibrosis (CF), a novel set of substituted polylysines were employed. Polylysine was partially neutralized by blocking a number of positively charged residues with gluconoyl groups. In addition, polylysine was substituted with sugar residues on a specified number of amino groups. Using the gluconoylated polylysine 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 μM chloroquine. Luciferase gene expression persisted at high levels for up to at least 120 hr following transfection. Glycosylated polylysines/pCMVLuc complexes were compared to the gluconoylated polylysine/pCMVLuc complex and β-Gal-, α-Glc-, and Lac-substituted polylysines gave 320%, 300%, and 290%, respectively, higher expression of the reporter gene luciferase. Luciferase expression ranged from 35 to 2 ng of luciferase per milligram of cell protein in the order: β-Gal = α-Glc = Lac > α-Gal = Rha = Man > β-GalNAc > α-GalNAc = α-Fuc, suggesting that 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 are attractive nonviral approach for the transfer of genes into airway epithelial cells.

Original languageEnglish (US)
Pages (from-to)1577-1586
Number of pages10
JournalHuman gene therapy
Volume7
Issue number13
DOIs
StatePublished - Aug 20 1996
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Molecular Biology
  • Genetics

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