Department of Medicine, Division of Infectious Diseases and Immunology
Adjuvants, Immunologic; Animals; Base Sequence; HEK293 Cells; Humans; Mice; MicroRNAs; Nucleotide Motifs; Oligonucleotides; RNA; Toll-Like Receptor 7; Toll-Like Receptor 8
Biochemistry | Immunology and Infectious Disease | Nucleic Acids, Nucleotides, and Nucleosides
Anti-microRNA (miRNA) oligonucleotides (AMOs) with 2'-O-Methyl (2'OMe) residues are commonly used to study miRNA function and can achieve high potency, with low cytotoxicity. Not withstanding this, we demonstrate the sequence-dependent capacity of 2'OMe AMOs to inhibit Toll-like receptor (TLR) 7 and 8 sensing of immunostimulatory RNA, independent of their miRNA-targeting function. Through a screen of 29 AMOs targeting common miRNAs, we found a subset of sequences highly inhibitory to TLR7 sensing in mouse macrophages. Interspecies conservation of this inhibitory activity was confirmed on TLR7/8 activity in human peripheral blood mononuclear cells. Significantly, we identified a core motif governing the inhibitory activity of these AMOs, which is present in more than 50 AMOs targeted to human miRNAs in miRBaseV20. DNA/locked nucleic acids (LNA) AMOs synthesized with a phosphorothioate backbone also inhibited TLR7 sensing in a sequence-dependent manner, demonstrating that the off-target effects of AMOs are not restricted to 2'OMe modification. Taken together, our work establishes the potential for off-target effects of AMOs on TLR7/8 function, which should be taken into account in their therapeutic development and in vivo application.
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Copyright The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
DOI of Published Version
Nucleic Acids Res. 2015 Jan;43(2):1177-88. doi: 10.1093/nar/gku1343. Epub 2014 Dec 24. Link to article on publisher's site.
Nucleic acids research
Sarvestani ST, Stunden HJ, Behlke MA, Forster SC, McCoy CE, Tate MD, Ferrand J, Lennox KA, Latz E, Williams BR, Gantier MP. (2015). Sequence-dependent off-target inhibition of TLR7/8 sensing by synthetic microRNA inhibitors. Open Access Articles. https://doi.org/10.1093/nar/gku1343. Retrieved from https://escholarship.umassmed.edu/oapubs/2578
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.