Lactosylated poly-l-lysine targets a potential lactose receptor in cystic fibrosis and non-cystic fibrosis airway epithelial cells
| Autor: | Qian-Chun Yu, Mary Catherine Glick, Daniel T. Klink, Thomas F. Scanlin |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
DNA
Complementary Cystic Fibrosis Endosome media_common.quotation_subject Lactose Endocytosis complex mixtures Clathrin Cell membrane Complementary DNA Drug Discovery Genetics medicine Humans Polylysine Internalization Molecular Biology Cells Cultured media_common Pharmacology biology Coated Pits Cell-Membrane Epithelial Cells Receptor-mediated endocytosis Transfection Molecular biology Trachea Microscopy Electron medicine.anatomical_structure biology.protein bacteria Molecular Medicine |
| Zdroj: | Molecular Therapy. 7:73-80 |
| ISSN: | 1525-0016 |
| DOI: | 10.1016/s1525-0016(02)00016-3 |
| Popis: | Poly-L-lysine with 40% of the epsilon -amino groups substituted with lactosyl residues facilitated the internalization of lactosylated poly-L-lysine/cDNA complexes into cystic fibrosis (CF) and non-CF airway epithelial cells. It was previously shown that lactosylated poly-L-lysine enhanced the transfer of cDNA into the cell nucleus, resulting in transfection. The cell entry of lactosylated poly-L-lysine/cDNA complexes, however, has not been elucidated and we hypothesized that entry of the complex was by receptor-mediated endocytosis. It is shown here that binding of the vector/cDNA complexes to the cell membrane was inhibited by lactose but not N-acetyl glucosamine. Examination by electron microscopy revealed the complexes in clathrin-coated pits. Furthermore, the complexes colocalized with transferrin during cell entry and were shown in early endosomes. These results demonstrated that lactosylated poly-L-lysine/cDNA complexes enter airway epithelial cells via receptor-mediated endocytosis utilizing lactose-binding receptors, which employ the clathrin-coated pit for internalization. Taken together with the fact that nuclear translocation also is enhanced by lactose, these results demonstrate why lactosylated poly-L-lysine is an excellent vector for transfection of airway epithelial cells. Moreover, other carbohydrates covalently linked to poly-L-lysine for targeting other specific cell types, combined with lactosyl residues, can be designed for the development of other molecular conjugates for gene transfer. |
| Databáze: | OpenAIRE |
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