TY - JOUR
T1 - 5-Ethynyluridine
T2 - A Bio-orthogonal Uridine Variant for mRNA-Based Therapies and Vaccines
AU - Maassen, Sjors
AU - Coenen, Britt
AU - Dulk, Sara
AU - van der Werff, Martijn
AU - Warner, Harry
AU - Spada, Fabio
AU - Frischmuth, Thomas
AU - Incarnato, Danny
AU - van den Bogaart, Geert
N1 - Funding Information:
This work was supported by the European Research Council (ERC) under the European Union's Horizon 2020 research innovation programme [grant agreement no. 862137] to G.v.d.B.; ZonMW [project grant no. 09120011910001] to G.v.d.B. The authors would like to acknowledge Pieter Grijpstra for his very competent management of lab utilities during the COVID‐19 pandemic to assure completion of this project.
Publisher Copyright:
© 2023 The Authors. ChemBioChem published by Wiley-VCH GmbH.
PY - 2023/3/1
Y1 - 2023/3/1
N2 - The identification of pseudo- and N1-methylpseudo-uridine (Ψ and mΨ, respectively) as immunosilent uridine analogues has propelled the development of mRNA-based vaccines and therapeutics. Here, we have characterised another uridine analogue, 5-ethynyluridine (EU), which has an ethynyl moiety. We show that this uridine analogue does not cause immune activation in human macrophages, as it does not induce interleukin-6 secretion or expression of the inflammatory and antiviral genes MX1, PKR, and TAP2. Moreover, EU allows for prolonged expression, as shown with mRNA coding for yellow fluorescent protein (YFP). Side-by-side comparisons of EU with unmodified, Ψ, and mΨ revealed that EU-modified mRNA is expressed at lower levels, but confers similar stability and low immunogenicity to the other uridine analogues. Furthermore, structure analysis of modified mRNAs suggests that the observed phenotype is largely independent of RNA folding. Thus, EU is a potential candidate for RNA-based vaccines and therapeutics.
AB - The identification of pseudo- and N1-methylpseudo-uridine (Ψ and mΨ, respectively) as immunosilent uridine analogues has propelled the development of mRNA-based vaccines and therapeutics. Here, we have characterised another uridine analogue, 5-ethynyluridine (EU), which has an ethynyl moiety. We show that this uridine analogue does not cause immune activation in human macrophages, as it does not induce interleukin-6 secretion or expression of the inflammatory and antiviral genes MX1, PKR, and TAP2. Moreover, EU allows for prolonged expression, as shown with mRNA coding for yellow fluorescent protein (YFP). Side-by-side comparisons of EU with unmodified, Ψ, and mΨ revealed that EU-modified mRNA is expressed at lower levels, but confers similar stability and low immunogenicity to the other uridine analogues. Furthermore, structure analysis of modified mRNAs suggests that the observed phenotype is largely independent of RNA folding. Thus, EU is a potential candidate for RNA-based vaccines and therapeutics.
KW - macrophage
KW - mRNA therapy
KW - uridine analogues
KW - viral immunity
UR - http://www.scopus.com/inward/record.url?scp=85147102045&partnerID=8YFLogxK
U2 - 10.1002/cbic.202200658
DO - 10.1002/cbic.202200658
M3 - Article
C2 - 36594506
AN - SCOPUS:85147102045
SN - 1439-4227
VL - 24
JO - ChemBioChem
JF - ChemBioChem
M1 - 202200658
ER -