Targeting the coronavirus nucleocapsid protein through GSK-3 inhibition

• Published in: Proceedings of the National Academy of Sciences - PNAS 2021-10, Vol.118 (42), p.1-9
• Main Authors: Liu, Xiaolei, Verma, Anurag, Garcia, Gustavo, Ramage, Holly, Lucas, Anastasia, Myers, Rebecca L., Michaelson, Jacob J., Coryell, William, Kumar, Arvind, Charney, Alexander W., Kazanietz, Marcelo G., Rader, Daniel J., Ritchie, Marylyn D., Berrettini, Wade H., Schultz, David C., Cherry, Sara, Damoiseaux, Robert, Arumugaswami, Vaithilingaraja, Klein, Peter S.
• Format: Article
• Language: English
• Subjects: Adult; Aged; Biological Sciences; Coronaviridae; Coronavirus Nucleocapsid Proteins - metabolism; Coronaviruses; COVID-19; COVID-19 - prevention & control; Epithelial cells; Epithelium; Female; Glycogen; Glycogen synthase kinase 3; Glycogen Synthase Kinase 3 - antagonists & inhibitors; Glycogen Synthase Kinase 3 - metabolism; Glycogens; HEK293 Cells; Humans; Kinases; Lithium; Lithium Compounds - pharmacology; Lithium Compounds - therapeutic use; Male; Middle Aged; Molecular Targeted Therapy; N protein; Nucleocapsids; Phosphoproteins - metabolism; Phosphorylation; Phosphorylation - drug effects; Proteins; Replication; Respiratory diseases; Retrospective Studies; Risk management; Severe acute respiratory syndrome coronavirus 2; Transcription; Viral diseases; Virions
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.2113401118

The coronaviruses responsible for severe acute respiratory syndrome (SARS-CoV), COVID-19 (SARS-CoV-2), Middle East respiratory syndrome-CoV, and other coronavirus infections express a nucleocapsid protein (N) that is essential for viral replication, transcription, and virion assembly. Phosphorylation of N from SARS-CoV by glycogen synthase kinase 3 (GSK-3) is required for its function and inhibition of GSK-3 with lithium impairs N phosphorylation, viral transcription, and replication. Here we report that the SARS-CoV-2 N protein contains GSK-3 consensus sequences and that this motif is conserved in diverse coronaviruses, raising the possibility that SARS-CoV-2 may be sensitive to GSK-3 inhibitors, including lithium. We conducted a retrospective analysis of lithium use in patients from three major health systems who were PCR-tested for SARS-CoV-2. We found that patients taking lithium have a significantly reduced risk of COVID-19 (odds ratio = 0.51 [0.35–0.74], P = 0.005). We also show that the SARS-CoV-2 N protein is phosphorylated by GSK-3. Knockout of GSK3A and GSK3B demonstrates that GSK-3 is essential for N phosphorylation. Alternative GSK-3 inhibitors block N phosphorylation and impair replication in SARS-CoV-2 infected lung epithelial cells in a cell-type–dependent manner. Targeting GSK-3 may therefore provide an approach to treat COVID-19 and future coronavirus outbreaks.