Bioreducible BPEI-SS-PEG-cNGR polymer as a tumor targeted nonviral gene carrier.
The work demonstrated development of multifunctional gene carrier which has incorporated reducible moiety, tumor targeting ligands as well as PEG to achieve efficient release of pDNA, enhanced tumor-specificity and long circulation, respectively. In our successful one-pot synthesis of multifunctional polymer, low molecular weight branched polyethylenimine (BPEI) was thiolated with propylene sulfide, and mixed with alpha-Maleimide-omega-N-hydroxysuccinimide ester polyethylene glycol (MAL-PEG-NHS, MW: 5000), and cyclic NGR peptide. The structural elucidation of the cNGR conjugated reducible BPEI containing disulfide bond (BPEI-SS-PEG-cNGR), was done by NMR and GPC study. Complex formation as well as reducible property of the polymer was confirmed by gel retardation assay. In order to achieve efficient tumor targeting, we have used cNGR peptide which is known to bind to CD13 overexpressed in neovasculature endothelial cells. Tumor target-specificity of polymer was established by carrying out competitive inhibition assay with free cNGR peptide. Cellular uptake of polymers was evaluated by confocal laser scanning microscope (CLSM). Finally, addition of free cNGR and buthionine sulfoximine (BSO) reduced transfection efficiency synergistically, which implied that multifunctional polymer-mediated gene transfection took place tumor-specifically and via GSH-dependent pathway.
Department of Chemistry, BK21 program, Polymer Research Institute, Pohang University of Science and Technology (POSTECH), San 31, Hyoja-dong, Nam-Gu, Pohang 790-784, Republic of Korea.
Sejin
S
Son
Kaushik
K
Singha
Won Jong
WJ
Kim
D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry*; D016923:Cell Death / Q000187:drug effects; D002460:Cell Line; D003432:Cross-Linking Reagents / Q000737:chemistry / Q000494:pharmacology; D004247:DNA / Q000235:genetics; D004220:Disulfides / Q000378:metabolism*; D018014:Gene Transfer Techniques; D006801:Humans; D051379:Mice; D009369:Neoplasms / Q000378:metabolism*; D009928:Organ Specificity / Q000187:drug effects; D010316:Particle Size; D010456:Peptides, Cyclic / Q000138:chemical synthesis / Q000737:chemistry*; D010957:Plasmids / Q000235:genetics; D011092:Polyethylene Glycols / Q000138:chemical synthesis / Q000737:chemistry*; D011094:Polyethyleneimine / Q000138:chemical synthesis / Q000737:chemistry*; D014162:Transfection; D014780:Viruses / Q000235:genetics
D001672:Biocompatible Materials; D003432:Cross-Linking Reagents; D004220:Disulfides; D010456:Peptides, Cyclic; D011092:Polyethylene Glycols; D011094:Polyethyleneimine; D004247:DNA