Inhibition of SARS-CoV-2 growth in the lungs of mice by a peptide-conjugated morpholino oligomer targeting viral RNA

Inhibition of SARS-CoV-2 growth in the lungs of mice by a peptide-conjugated morpholino oligomer targeting viral RNA

Molecular Therapy - Nucleic Acids

10.1016/j.omtn.2024.102331

https://doi.org/10.1016/j.omtn.2024.102331

Alexandra Sakai, Gagandeep Singh, Mahsa Khoshbakht, Scott Bittner, Christiane V. Löhr, Randy Diaz-Tapia, Prajakta Warang, Kris White, Luke Le Luo, Blanton Tolbert, Mario Blanco, Amy Chow, Mitchell Guttman, Cuiping Li, Yiming Bao, Joses Ho, Sebastian Maurer-Stroh, Arnab Chatterjee, Sumit Chanda, Adolfo García-Sastre, Michael Schotsaert, John R. Teijaro, Hong M. Moulton, David A. Stein

MT: Oligonucleotides, Therapies and applications

10 September 2024

Sakai, A., Singh, G., Khoshbakht, M., Bittner, S., Löhr, C. V., Diaz-Tapia, R., Warang, P., White, K., Luo, L. L., Tolbert, B., Blanco, M., Chow, A., Guttman, M., Li, C., Bao, Y., Ho, J., Maurer-Stroh, S., Chatterjee, A., Chanda, S., … Stein, D. A. (2024). Inhibition of SARS-CoV-2 growth in the lungs of mice by a peptide-conjugated morpholino oligomer targeting viral RNA. Molecular Therapy - Nucleic Acids, 35(4), 102331. https://doi.org/10.1016/j.omtn.2024.102331

Further development of direct-acting antiviral agents against human SARS-CoV-2 infections remains a public health priority. Here, we report that an antisense peptide-conjugated morpholino oligomer (PPMO) named 5′END-2, targeting a highly conserved sequence in the 5′ UTR of SARS-CoV-2 genomic RNA, potently suppressed SARS-CoV-2 growth in vitro and in vivo. In HeLa-ACE 2 cells, 5′END-2 produced IC50 values of between 40 nM and 1.15 μM in challenges using six genetically disparate strains of SARS-CoV-2, including JN.1. In vivo, using K18-hACE2 mice and the WA-1/2020 virus isolate, two doses of 5′END-2 at 10 mg/kg, administered intranasally on the day before and the day after infection, produced approximately 1.4 log10 virus titer reduction in lung tissue at 3 days post-infection. Under a similar dosing schedule, intratracheal administration of 1.0–2.0 mg/kg 5′END-2 produced over 3.5 log10 virus growth suppression in mouse lungs. Electrophoretic mobility shift assays characterized specific binding of 5′END-2 to its complementary target RNA. Furthermore, using reporter constructs containing SARS-CoV-2 5′ UTR leader sequence, in an in-cell system, we observed that 5′END-2 could interfere with translation in a sequence-specific manner. The results demonstrate that direct pulmonary delivery of 5′END-2 PPMO is a promising antiviral strategy against SARS-CoV-2 infections and warrants further development.

Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)

This research / project is supported by the National Institute of Allergy and Infectious Diseases (NIAID) Antiviral Drug Discovery program, - NA
Grant Reference no. : U19AI171443

This research / project is supported by the NIH/National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) - NA
Grant Reference no. : NIH/NIAID R01AI160706 / R01DK130425

https://oar.a-star.edu.sg/communities-collections/articles/22199

2162-2531

Bioinformatics Institute