2'-O-Methyl-guanosine 3-base RNA fragments mediate essential natural TLR7/8 antagonism

Alharbi, A.; Sapkota, S.; Zhang, Z.; Jin, R.; Jayasekara, W. S.; Rupasinghe, E.; Speir, M.; Wilkinson-White, L.; Gamsjaeger, R.; Cubeddu, L.; Ellyard, J. I.; Wenholz, D.; McAllan, A.; Rezwan, R.; Ying, L.; Hosseini Far, H.; Bones, J.; He, S.; Yu, D.; Lennox, K.; Hertzog, P. J.; Vinuesa, C.; Behlke, M. A.; Ohto, U.; Laczka, O.; Corry, B.; Shimizu, T.; Gantier, M. P.

doi:10.1101/2024.07.25.605091

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bioRxiv · 10.1101/2024.07.25.605091

2'-O-Methyl-guanosine 3-base RNA fragments mediate essential natural TLR7/8 antagonism

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Abstract

Recognition of RNA fragments by Toll-like receptors (TLR) 7 and 8 is a key contributor to the initiation of a protective innate immune response against pathogens. A long-standing enigma is how degradation products of host RNAs, generated by the daily phagocytic clearance of billions of apoptotic cells, fail to activate TLR7 and TLR8 signalling1. Here, we report that select 2-O-methyl (2-Ome) guanosine RNA fragments as short as 3 bases, including those derived from host-RNAs, are potent TLR7 and TLR8 antagonists that reduce TLR7 sensing in vivo. Mechanistically, antagonistic fragments are directed towards a distinct binding site on these proteins by 5-end 2-Ome guanosine. Our results indicate that host-RNAs evade detection by TLR7/8 due to a pool of abundant host ribosomal 2-Ome-modified RNA fragments that naturally antagonize TLR7 and TLR8 sensing to avoid auto-immunity. Crucially, rare TLR7 and TLR8 mutations located at this antagonistic site decrease the inhibitory activity of 2-Ome guanosine RNA fragments and lead to auto-immunity in patients. Our findings also establish that select chemically synthesised 3-base oligonucleotides can harness the protective anti-inflammatory activity of this natural immune checkpoint for therapeutic targeting of TLR7-driven diseases. One Sentence SummaryShort 2-O-Methyl RNA fragments are natural TLR7/8 antagonists

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Alharbi, A., Sapkota, S., Zhang, Z., Jin, R., Jayasekara, W. S., Rupasinghe, E., Speir, M., Wilkinson-White, L., Gamsjaeger, R., Cubeddu, L., Ellyard, J. I., Wenholz, D., McAllan, A., Rezwan, R., Ying, L., Hosseini Far, H., Bones, J., He, S., Yu, D., Lennox, K., Hertzog, P. J., Vinuesa, C., Behlke, M. A., Ohto, U., Laczka, O., Corry, B., Shimizu, T., Gantier, M. P.. 2024-07-25. 2'-O-Methyl-guanosine 3-base RNA fragments mediate essential natural TLR7/8 antagonism. https://doi.org/10.1101/2024.07.25.605091

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