N4-hydroxycytidine, the active compound of Molnupiravir, promotes SARS-CoV-2 mutagenesis and escape from a neutralizing nanobody
Arne Zibat,
Xiaoxiao Zhang,
Antje Dickmanns,
Kim M. Stegmann,
Adrian W. Dobbelstein,
Halima Alachram,
Rebecca Soliwoda,
Gabriela Salinas,
Uwe Groß,
Dirk Görlich,
Maik Kschischo,
Bernd Wollnik,
Matthias Dobbelstein
Affiliations
Arne Zibat
Department of Human Genetics, University Medical Center Göttingen, 37073 Göttingen, Germany
Xiaoxiao Zhang
Department of Mathematics and Technology, University of Applied Sciences Koblenz, 53424 Remagen, Germany; Department of Informatics, Technical University of Munich, 81675 Munich, Germany
Antje Dickmanns
Department of Molecular Oncology, Göttingen Center of Molecular Biosciences (GZMB), University Medical Center Göttingen, 37077 Göttingen, Germany
Kim M. Stegmann
Department of Molecular Oncology, Göttingen Center of Molecular Biosciences (GZMB), University Medical Center Göttingen, 37077 Göttingen, Germany
Adrian W. Dobbelstein
Max Planck Institute for Biology, 72076 Tübingen, Germany
Halima Alachram
Department of Human Genetics, University Medical Center Göttingen, 37073 Göttingen, Germany
Rebecca Soliwoda
Department of Molecular Oncology, Göttingen Center of Molecular Biosciences (GZMB), University Medical Center Göttingen, 37077 Göttingen, Germany
Gabriela Salinas
NGS Integrative Genomics Core Unit, Department of Human Genetics, University Medical Center Göttingen, 37077 Göttingen, Germany
Uwe Groß
Department of Medical Microbiology and Virology, University Medical Center Göttingen, 37075 Göttingen, Germany
Dirk Görlich
Max Planck Institute for Multidisciplinary Sciences, 37077 Göttingen, Germany
Maik Kschischo
Department of Mathematics and Technology, University of Applied Sciences Koblenz, 53424 Remagen, Germany
Bernd Wollnik
Department of Human Genetics, University Medical Center Göttingen, 37073 Göttingen, Germany; Cluster of Excellence ''Multiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells'' (MBExC), University of Göttingen, 37075 Göttingen, Germany
Matthias Dobbelstein
Department of Molecular Oncology, Göttingen Center of Molecular Biosciences (GZMB), University Medical Center Göttingen, 37077 Göttingen, Germany; Corresponding author
Summary: N4-hydroxycytidine (NHC), the active compound of the drug Molnupiravir, is incorporated into SARS-CoV-2 RNA, causing false base pairing. The desired result is an “error catastrophe,” but this bears the risk of mutated virus progeny. To address this experimentally, we propagated the initial SARS-CoV-2 strain in the presence of NHC. Deep sequencing revealed numerous NHC-induced mutations and host-cell-adapted virus variants. The presence of the neutralizing nanobody Re5D06 selected for immune escape mutations, in particular p.E484K and p.F490S, which are key mutations of the Beta/Gamma and Omicron-XBB strains, respectively. With NHC treatment, nanobody resistance occurred two passages earlier than without. Thus, within the limitations of this purely in vitro study, we conclude that the combined action of Molnupiravir and a spike-neutralizing antagonist leads to the rapid emergence of escape mutants. We propose caution use and supervision when using Molnupiravir, especially when patients are still at risk of spreading virus.
