Inhibition of multiple subtypes of influenza A virus in cell cultures with morpholino oligomers

Authors

Qing Ge
Manoj Pastey
Darwyn Kobasa
Piliapan Puthavathana
Christopher LupferFollow
Richard K. Bestwick
Patrick L. Iversen
Jianzhu Chen
David A. Stein

Abstract

Peptide-conjugated phosphorodiamidate morpholino oligomers (P-PMO) are single-stranded nucleic acid-like antisense agents that can reduce gene expression by sterically blocking complementary RNA sequence. P-PMO are water soluble and nuclease resistant, and they readily achieve uptake into cells in culture under standard conditions. Eight P-PMO, each 20 to 22 bases in length, were evaluated for their ability to inhibit influenza A virus (FLUAV) A/PR/8/34 (H1N1) replication in cell culture. The P-PMO were designed to base pair with FLUAV RNA sequences that are highly conserved across viral subtypes and considered critical to the FLUAV biological-cycle, such as gene segment termini and mRNA translation start site regions. Several P-PMO were highly efficacious, each reducing viral titer in a dose-responsive and sequence-specific manner in A/PR/8/34-infected cells. Two P-PMO, one designed to target the AUG translation start site region of PB1 mRNA and the other the 3′-terminal region of nucleoprotein viral genome RNA, also proved to be potent against several other FLUAV strains, including A/WSN/33 (H1N1), A/Memphis/8/88 (H3N2), A/Eq/Miami/63 (H3N8), A/Eq/Prague/56 (H7N7), and the highly pathogenic A/Thailand/1(KAN-1)/04 (H5N1). The P-PMO exhibited minimal cytotoxicity in cell viability assays. High efficacy by two of the P-PMO against multiple FLUAV subtypes suggests that these oligomers represent a broad-spectrum therapeutic approach against a high percentage of known FLUAV strains.

Document Type

Article

DOI

https://doi.org/10.1128/AAC.00644-06

Publication Date

11-1-2006

Recommended Citation

Ge, Qing, Manoj Pastey, Darwyn Kobasa, Piliapan Puthavathana, Christopher Lupfer, Richard K. Bestwick, Patrick L. Iversen, Jianzhu Chen, and David A. Stein. "Inhibition of multiple subtypes of influenza A virus in cell cultures with morpholino oligomers." Antimicrobial agents and chemotherapy 50, no. 11 (2006): 3724-3733.

Journal Title

Antimicrobial Agents and Chemotherapy