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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Citation: 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.
DOI of Published Version
Nucleic acids research
Sarvestani, Soroush T.; Stunden, H. James; Behlke, Mark A.; Forster, Samuel C.; McCoy, Claire E.; Tate, Michelle D.; Ferrand, Jonathan; Lennox, Kim A.; Latz, Eicke; Williams, Bryan R. G.; and Gantier, Michael P., "Sequence-dependent off-target inhibition of TLR7/8 sensing by synthetic microRNA inhibitors" (2015). Open Access Articles. 2578.
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