Development of modified siRNA molecules incorporating 5-fluoro-2'-deoxyuridine residues to enhance cytotoxicity
| Autor: | William H. Gmeiner, Tian Min Chen, Edward Chu, Shao yu Wu, John C. Schmitz |
|---|---|
| Rok vydání: | 2013 |
| Předmět: |
Small interfering RNA
Apoptosis Biology Transfection Thymidylate synthase 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Cell Line Tumor Genetics Humans Nucleotide RNA Small Interfering Cytotoxicity Ternary complex 030304 developmental biology chemistry.chemical_classification 0303 health sciences Thymidylate Synthase Deoxyuridine 3. Good health chemistry Biochemistry 030220 oncology & carcinogenesis biology.protein RNA Nucleoside |
| Zdroj: | Nucleic Acids Research |
| ISSN: | 1362-4962 0305-1048 |
| DOI: | 10.1093/nar/gkt120 |
| Popis: | Therapeutic small interfering RNAs (siRNAs) are composed of chemically modified nucleotides, which enhance RNA stability and increase affinity in Watson-Crick base pairing. However, the precise fate of such modified nucleotides once the siRNA is degraded within the cell is unknown. Previously, we demonstrated that deoxythymidine release from degraded siRNAs reversed the cytotoxicity of thymidylate synthase (TS)-targeted siRNAs and other TS inhibitor compounds. We hypothesized that siRNAs could be designed with specific nucleoside analogues that, once released, would enhance siRNA cytotoxicity. TS-targeted siRNAs were designed that contained 5-fluoro-2'-deoxyuridine (FdU) moieties at various locations within the siRNA. After transfection, these siRNAs suppressed TS protein and messenger RNA expression with different efficiencies depending on the location of the FdU modification. FdU was rapidly released from the siRNA as evidenced by formation of the covalent inhibitory ternary complex formed between TS protein and the FdU metabolite, FdUMP. These modified siRNAs exhibited 10-100-fold greater cytotoxicity and induced multiple DNA damage repair and apoptotic pathways when compared with control siRNAs. The strategy of designing siRNA molecules that incorporate cytotoxic nucleosides represents a potentially novel drug development approach for the treatment of cancer and other human diseases. |
| Databáze: | OpenAIRE |
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