1. Improved gene transfer to neuroblastoma cells by a monoclonal antibody targeting RET, a receptor tyrosine kinase.
- Author
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Yano L, Shimura M, Taniguchi M, Hayashi Y, Suzuki T, Hatake K, Takaku F, and Ishizaka Y
- Subjects
- Avidin chemistry, DNA chemistry, Extracellular Matrix metabolism, Genes, Reporter, Humans, Neuroblastoma genetics, Neuroblastoma immunology, Neurons metabolism, Pheochromocytoma genetics, Pheochromocytoma immunology, Pheochromocytoma therapy, Polylysine chemistry, Proto-Oncogene Proteins metabolism, Proto-Oncogene Proteins c-ret, Receptor Protein-Tyrosine Kinases metabolism, Recombinant Proteins genetics, Recombinant Proteins immunology, Recombinant Proteins metabolism, Transfection, Tumor Cells, Cultured, beta-Galactosidase genetics, beta-Galactosidase metabolism, Antibodies, Monoclonal pharmacology, Drosophila Proteins, Gene Transfer Techniques, Neuroblastoma therapy, Proto-Oncogene Proteins genetics, Proto-Oncogene Proteins immunology, Receptor Protein-Tyrosine Kinases genetics, Receptor Protein-Tyrosine Kinases immunology
- Abstract
Receptor-mediated gene transfer is an effective strategy among nonviral vector systems. It is, however, crucial to develop various types of monoclonal antibodies satisfying both the binding specificity for cell targeting and the capacity of endocytosis required for gene transfer. In the present study, we generated a novel monoclonal antibody (NBL-1) to RET, a receptor tyrosine kinase expressed in both neuroblastoma cells and cells present in substantia nigra, a responsive locus of Parkinson's disease. NBL-1, when added to the culture medium of the neuroblastoma cells, was incorporated by endocytosis in a wortmannin-sensitive manner. Using a biotinylated NBL-1 complexed with plasmid DNAs based on electrostatic interaction through avidin-conjugated polylysines, exogenous luciferase genes were expressed in neuroblastoma cells at a more than 10-fold higher level. The expression level of the gene based on NBL-1 was comparable to that obtained by a geneporter system, an improved nonviral gene transduction method. Furthermore, the NBL-1-based gene transfer mediated the formation of more than 20-fold higher numbers of drug-resistant colonies. In contrast, RET-negative cells, which included HeLa, HT1080, Caco-2, and Colo205 cells, did not show any increased expression of an exogenous gene by NBL-1. These data suggest that the RET molecules enable selective gene transduction, and that NBL-1 may possibly be applied to gene therapy for neuroblastomas and Parkinson's disease.
- Published
- 2000
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