2'-O-(2-Methoxyethyl) Nucleosides Are Not Phosphorylated or Incorporated Into the Genome of Human Lymphoblastoid TK6 Cells
| Autor: | Nicholas Edmunds, Amer F. Saleh, Catherine C. Priestley, Martin Bachman, Patrik U. Andersson, Mick D. Fellows, Nigel J. Gooderham, Scott P. Henry |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
DNA incorporation nucleoside kinase modified nucleosides Toxicology Thymidine Kinase Substrate Specificity 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Cell Line Tumor Deoxycytidine Kinase medicine Humans Nucleotide nucleoside phosphorylation Phosphorylation Nucleotide salvage chemistry.chemical_classification Cell Nucleus Nucleoside analogue Oligonucleotide Genome Human RNA Nucleosides Deoxycytidine kinase DNA 030104 developmental biology chemistry Biochemistry Thymidine kinase 030220 oncology & carcinogenesis 1115 Pharmacology And Pharmaceutical Sciences medicine.drug |
| Zdroj: | Toxicological sciences : an official journal of the Society of Toxicology. 163(1) |
| ISSN: | 1096-0929 |
| Popis: | Nucleoside analogs with 2'-modified sugar moieties are often used to improve the RNA target affinity and nuclease resistance of therapeutic oligonucleotides in preclinical and clinical development. Despite their enhanced nuclease resistance, oligonucleotides could slowly degrade releasing nucleoside analogs that have the potential to become phosphorylated and incorporated into cellular DNA and RNA. For the first time, the phosphorylation and DNA/RNA incorporation of 2'-O-(2-methoxyethyl) (2'-O-MOE) nucleoside analogs have been investigated. Using liquid chromatography/tandem mass spectrometry, we showed that enzymes in the nucleotide salvage pathway including deoxycytidine kinase (dCK) and thymidine kinase (TK1) displayed poor reactivity toward 2'-O-MOE nucleoside analogs. On the other hand, 2'-fluoro (F) nucleosides, regardless of the nucleobase, were efficiently phosphorylated to their monophosphate forms by dCK and TK1. Consistent with their efficient phosphorylation by dCK and TK1, 2'-F nucleoside analogs were incorporated into cellular DNA and RNA while no incorporation was detected with 2'-O-MOE nucleoside analogs. In conclusion, these data suggest that the inability of dCK and TK1 to create the monophosphates of 2'-O-MOE nucleoside analogs reduces the risk of their incorporation into cellular DNA and RNA. |
| Databáze: | OpenAIRE |
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