Targeted Lysosome Disruptive Elements for Improvement of Parenchymal Liver Cell-specific Gene Delivery

• Published in: The Journal of biological chemistry 2002-11, Vol.277 (48), p.45803-45810
• Main Authors: Van Rossenberg, Sabine M W, Sliedregt-Bol, Karen M, Meeuwenoord, Nico J, Van Berkel, Theo J C, Van Boom, Jacques H, Van Der Marel, Gijs A, Biessen, Erik A L
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Gene Transfer Techniques; In Vitro Techniques; Liver - cytology; Liver - metabolism; Lysosomes - metabolism; Male; Mice; Mice, Inbred C57BL; Molecular Sequence Data
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M203510200

The transfection ability of nonviral gene therapy vehicles is generally hampered by untimely lysosomal degradation of internalized DNA. In this study we describe the development of a targeted lysosome disruptive element to facilitate the escape of DNA from the lysosomal compartment, thus enhancing the transfection efficacy, in a cell-specific fashion. Two peptides (INF7 and JTS-1) were tested for their capacity to disrupt liposomes. In contrast to JTS-1, INF7 induced rapid cholesterol-independent leakage (EC 50 , 1.3 μ m ). INF7 was therefore selected for coupling to a high affinity ligand for the asialoglycoprotein receptor (ASGPr), K(GalNAc) 2 , to im- prove its uptake by parenchymal liver cells. Although the parent peptide disrupted both cholesterol-rich and -poor liposomes, the conjugate, INF7-K(GalNAc) 2 , only induced leakage of cholesterol-poor liposomes. Given that endosomal membranes of eukaryotic cells contain