2. Academic Validation
  3. Cytidine acetylation yields a hypoinflammatory synthetic messenger RNA

Cytidine acetylation yields a hypoinflammatory synthetic messenger RNA

  • Cell Chem Biol. 2022 Feb 17;29(2):312-320.e7. doi: 10.1016/j.chembiol.2021.07.003.
Kellie D Nance 1,  Supuni Thalalla Gamage 1,  Md Masud Alam 2,  Acong Yang 3,  Michaella J Levy 4,  Courtney N Link 1,  Laurence Florens 4,  Michael P Washburn 5,  Shuo Gu 3,  Joost J Oppenheim 2,  Jordan L Meier 6
Affiliations

Affiliations

  • 1 Chemical Biology Laboratory, Center for Cancer Research, National Cancer Institute, National Institutes of Health, 538 Chandler Street, Frederick, MD 21702, USA.
  • 2 Laboratory of Cancer Immunometabolism, Center for Cancer Research, National Cancer Institute, National Institutes of Health, 538 Chandler Street, Frederick, MD 21702, USA.
  • 3 RNA Biology Laboratory, Center for Cancer Research, National Cancer Institute, National Institutes of Health, 538 Chandler Street, Frederick, MD 21702, USA.
  • 4 Stowers Institute for Medical Research, Kansas City, MO 64110, USA.
  • 5 Department of Cancer Biology, University of Kansas Medical Center, Kansas City, KS 66160, USA.
  • 6 Chemical Biology Laboratory, Center for Cancer Research, National Cancer Institute, National Institutes of Health, 538 Chandler Street, Frederick, MD 21702, USA. Electronic address: [email protected].
PMID: 35180432 DOI: 10.1016/j.chembiol.2021.07.003
Abstract

Synthetic messenger RNA (mRNA) is an emerging therapeutic platform with important applications in oncology and infectious disease. Effective mRNA medicines must be translated by the ribosome but not trigger a strong nucleic acid-mediated immune response. To expand the medicinal chemistry toolbox for these agents, here we report the properties of the naturally occurring nucleobase N4-acetylcytidine (ac4C) in synthetic mRNAs. We find that ac4C is compatible with, but does not enhance, protein production in the context of synthetic mRNA reporters. However, replacement of cytidine with ac4C diminishes inflammatory gene expression in immune cells caused by synthetic mRNAs. Chemoproteomic capture indicates that ac4C alters the protein interactome of synthetic mRNAs, reducing binding to cytidine-binding proteins and an immune sensor. Overall, our studies illustrate the unique ability of ac4C to modulate RNA-protein interactions and provide a foundation for using N4-cytidine acylation to fine-tune the properties of nucleic acid therapeutics.

Keywords

RNA modifications; acetylation; acetylcytidine; mRNA vaccines; messenger RNA; pseudouridine.

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