2'-O-(2-methoxyethyl) nucleosides are not phosphorylated or incorporated into the genome of Human Lymphoblastoid TK6 Cells

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Title: 2'-O-(2-methoxyethyl) nucleosides are not phosphorylated or incorporated into the genome of Human Lymphoblastoid TK6 Cells
Author(s): Saleh, AF
Bachman, M
Priestley, CC
Gooderham, NJ
Andersson, P
Henry, SP
Edmunds, N
Fellows, MD
Item Type: Journal Article
Abstract: Nucleoside analogues 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 analogues that have the potential to become phosphorylated and incorporated into cellular DNA and RNA. For the first time, the phosphorylation and DNA and RNA incorporation of 2'-O-(2-methoxyethyl) (2'-O-MOE) nucleoside analogues have been investigated. Using LC/MS/MS, 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 analogues. 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 nucleosides analogues were incorporated into cellular DNA and RNA while no incorporation was detected with 2'-O-MOE nucleoside analogues. In conclusion, these data suggest that the inability of dCK and TK1 to create the monophosphates of 2'-O-MOE nucleoside analogues reduces the risk of their incorporation into cellular DNA and RNA.
Publication Date: 9-Jan-2018
Date of Acceptance: 2-Jan-2018
URI: http://hdl.handle.net/10044/1/56042
DOI: https://dx.doi.org/10.1093/toxsci/kfy005
ISSN: 1096-0929
Publisher: Oxford University Press (OUP)
Start Page: 70
End Page: 78
Journal / Book Title: Toxicological Sciences
Volume: 163
Issue: 1
Copyright Statement: © The Author(s) 2018. Published by Oxford University Press on behalf of the Society of Toxicology. This is a pre-copyedited, author-produced PDF of an article accepted for publication in Toxicological Sciences following peer review. The version of record Amer F Saleh, Martin Bachman, Catherine C Priestley, Nigel J Gooderham, Patrik Andersson, Scott P Henry, Nick Edmunds, Mick D Fellows; 2′-O-(2-methoxyethyl) nucleosides are not phosphorylated or incorporated into the genome of Human Lymphoblastoid TK6 Cells, Toxicological Sciences, , kfy005 is available online at: https://dx.doi.org/10.1093/toxsci/kfy005
Keywords: DNA incorporation
modified nucleosides
nucleoside kinase
nucleoside phosphorylation
1115 Pharmacology And Pharmaceutical Sciences
Toxicology
Publication Status: Published
Embargo Date: 2019-01-09
Appears in Collections:Division of Surgery
Faculty of Medicine



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