title: Construction of an aptamer modified liposomal system targeted to tumor endothelial cells.
abstract: We describe herein the development of a high affinity and specific DNA aptamer as a new ligand for use in liposomal nanoparticles to target cultured mouse tumor endothelial cells (mTECs). Active targeted nanotechnology based drug delivery systems are currently of great interest, due to their potential for reducing side effects and facilitating the delivery of cytotoxic drugs or genes in a site specific manner. In this study, we report on a promising aptamer candidate AraHH036 that shows selective binding towards mTECs. The aptamer does not bind to normal cells, normal endothelial cells or tumor cells. Therefore, we synthesized an aptamer-polyethylene glycol (PEG) lipid conjugate and prepared aptamer based liposomes (ALPs) by the standard lipid hydration method. First, we quantified the higher capacity of ALPs to internalize into mTECs by incubating ALPs containing 1 mol%, 5 mol% and 10 mol% aptamer of total lipids and compared the results to those for unmodified PEGylated liposomes (PLPs). A confocal laser scanning microscope (CLSM) uptake study indicated that the ALPs were taken up more efficiently than PLPs. The measured Kd value of the ALPs was 142 nM. An intracellular trafficking study confirmed that most of the rhodamine labeled ALPs were taken up and co-localized with the green lysotracker, thus confirming that they were located in lysosomes. Finally, using an aptamer based proteomics approach, the molecular target protein of the aptamer was identified as heat shock protein 70 (HSP70). The results suggest that these ALPs offer promise as a new carrier molecule for delivering anti-angiogenesis drugs to tumor vasculature.
authors.affiliation: Laboratory of Innovative Nanomedicine, Faculty of Pharmaceutical Sciences, Hokkaido University.
authors.forename: Mst Naznin
authors.initials: MN
authors.lastname: Ara
authors.forename: Takashi
authors.initials: T
authors.lastname: Matsuda
authors.forename: Mamoru
authors.initials: M
authors.lastname: Hyodo
authors.forename: Yu
authors.initials: Y
authors.lastname: Sakurai
authors.forename: Noritaka
authors.initials: N
authors.lastname: Ohga
authors.forename: Kyoko
authors.initials: K
authors.lastname: Hida
authors.forename: Hideyoshi
authors.initials: H
authors.lastname: Harashima
mesh_terms: D000818:Animals; D052157:Aptamers, Nucleotide / Q000008:administration & dosage* / Q000737:chemistry; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000378:metabolism*; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D006801:Humans; D008081:Liposomes; D008247:Lysosomes / Q000378:metabolism; D008301:Maleimides / Q000737:chemistry; D051379:Mice; D008819:Mice, Nude; D041681:NIH 3T3 Cells; D053758:Nanoparticles; D009369:Neoplasms / Q000378:metabolism*; D010714:Phosphatidylethanolamines / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D040901:Proteomics; D012867:Skin / Q000166:cytology
chemical_list: D052157:Aptamers, Nucleotide; D018840:HSP70 Heat-Shock Proteins; D008081:Liposomes; D008301:Maleimides; D010714:Phosphatidylethanolamines; C038089:1,2-distearoylphosphatidylethanolamine; C043592:maleimide; D011092:Polyethylene Glycols