title: Cell-specific induction of apoptosis by rationally designed bivalent aptamer-siRNA transcripts silencing eukaryotic elongation factor 2.
abstract: New strategies for cell type-specific delivery need to be developed if RNA interference is to realize its full therapeutic potential. One possible approach is the use of aptamers to deliver siRNAs selectively to tumor cells with appropriate antigens displayed on the surface. We used an aptamer that binds specifically to PSMA, a cell surface glycoprotein found in abundance on prostate cancer cells, and joined its 3' end to a siRNA specific for Eukaryotic Elongation Factor 2 mRNA (EEF2). This is an attractive target for cancer therapy because inhibiting EEF2 causes the rapid arrest of protein synthesis, inducing apoptosis and leading ultimately to cell death. In order to enhance the therapeutic efficacy of the aptamer-siRNA, we increased the valency of the construct by rational design. Two anti-PSMA aptamers were designed such that each binding sequence could fold independently into its active conformation. Here we show specific cytotoxicity resulting from siRNA-induced silencing of EEF2, as well as specific delivery to PSMA-expressing prostate cancer cells. Increasing the valency of the aptamer resulted in enhanced cytotoxicity compared with the monovalent constructs. The results presented here demonstrate the usefulness of multivalent aptamer-based delivery vehicles for siRNA therapeutics.
authors.affiliation: Department of Pharmaceutical Product Development, Fraunhofer Institute for Molecular Biology and Applied Ecology, Aachen, Germany.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Wullner
authors.forename: Inga
authors.initials: I
authors.lastname: Neef
authors.forename: Andreas
authors.initials: A
authors.lastname: Eller
authors.forename: Michael
authors.initials: M
authors.lastname: Kleines
authors.forename: Mehmet Kemal
authors.initials: MK
authors.lastname: Tur
authors.forename: Stefan
authors.initials: S
authors.lastname: Barth
mesh_terms: D017209:Apoptosis / Q000235:genetics*; D052157:Aptamers, Nucleotide / Q000008:administration & dosage* / Q000138:chemical synthesis / Q000235:genetics*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000235:genetics; D015195:Drug Design; D054736:Elongation Factor 2 Kinase / Q000037:antagonists & inhibitors* / Q000235:genetics*; D020868:Gene Silencing; D018390:Gene Targeting / Q000379:methods; D006801:Humans; D011485:Protein Binding / Q000235:genetics; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000138:chemical synthesis / Q000235:genetics*; D014158:Transcription, Genetic
chemical_list: D052157:Aptamers, Nucleotide; D034741:RNA, Small Interfering; C517689:EEF2K protein, human; D054736:Elongation Factor 2 Kinase