The Critical Contribution of Pseudouridine to mRNA COVID-19 Vaccines

Abstract

The current COVID-19 pandemic is a massive source of global disruption, having led so far to two hundred and fifty million COVID-19 cases and almost five million deaths worldwide. It was recognized in the beginning that only an effective vaccine could lead to a way out of the pandemic, and therefore the race for the COVID-19 vaccine started immediately, boosted by the availability of the viral sequence data. Two novel vaccine platforms, based on mRNA technology, were developed in 2020 by Pfizer-BioNTech and Moderna Therapeutics (comirnaty® and spikevax®, respectively), and were the first ones presenting efficacies higher than 90%. Both consisted of N1-methyl-pseudouridine-modified mRNA encoding the SARS-COVID-19 Spike protein and were delivered with a lipid nanoparticle (LNP) formulation. Because the delivery problem of ribonucleic acids had been known for decades, the success of LNPs was quickly hailed by many as the unsung hero of COVID-19 mRNA vaccines. However, the clinical trial efficacy results of the Curevac mRNA vaccine (CVnCoV) suggested that the delivery system was not the only key to the success. CVnCoV consisted of an unmodified mRNA (encoding the same spike protein as Moderna and Pfizer-BioNTech's mRNA vaccines) and was formulated with the same LNP as Pfizer-BioNTech's vaccine (Acuitas ALC-0315). However, its efficacy was only 48%. This striking difference in efficacy could be attributed to the presence of a critical RNA modification (N1-methyl-pseudouridine) in the Pfizer-BioNTech and Moderna's mRNA vaccines (but not in CVnCoV). Here we highlight the features of N1-methyl-pseudouridine and its contributions to mRNA vaccines.

Keywords: COVID-19; N1-methyl-pseudouridine; RNA modification; SARS-CoV-2; lipid nanoparticles; mRNA; pseudouridine; vaccines.

FIGURE 1

Schematic representation of U-to-Ψ isomerization and additional N1 methylation. Ψ is a rotational isomer of uridine, in which the N-C glycosidic bond is substituted with the C-C bond. The isomerization reaction also creates an extra hydrogen bond donor (-N1H). Ψ can be further methylated at the N1 position by Nep1 (an N1-specific Ψ methyltransferase) to generate N-methyl-Ψ. d, hydrogen bond donor; a, hydrogen bond acceptor.

FIGURE 2

Schematics of SARS-COVID 19 mRNA vaccination. The vaccine consists of unmodified or N-methyl-Ψ-modified mRNA (encoding the SARS-COVID-19 spike protein) and lipid nanoparticles (LNPs). It is injected into the muscle of the upper arm to create an immune response. N-methyl-Ψ-modified mRNA exhibits higher efficacy (more than 90% of efficacy against COVID-19 symptoms) as compared to the unmodified mRNA vaccines (lower than 50%).