VP35 knockdown inhibits Ebola virus amplification and protects against lethal infection in mice.

Abstract

Phosphorodiamidate morpholino oligomers (PMO) are a class of uncharged single-stranded DNA analogs modified such that each subunit includes a phosphorodiamidate linkage and morpholine ring. PMO antisense agents have been reported to effectively interfere with the replication of several positive-strand RNA viruses in cell culture. The filoviruses, Marburg virus and Ebola virus (EBOV), are negative-strand RNA viruses that cause up to 90% lethality in human outbreaks. There is currently no commercially available vaccine or efficacious therapeutic for any filovirus. In this study, PMO conjugated to arginine-rich cell-penetrating peptide (P-PMO) and nonconjugated PMO were assayed for the ability to inhibit EBOV infection in cell culture and in a mouse model of lethal EBOV infection. A 22-mer P-PMO designed to base pair with the translation start site region of EBOV VP35 positive-sense RNA generated sequence-specific and time- and dose-dependent inhibition of EBOV amplification in cell culture. The same oligomer provided complete protection to mice when administered before or after an otherwise lethal infection of EBOV. A corresponding nonconjugated PMO, as well as nonconjugated truncated versions of 16 and 19 base residues, provided length-dependent protection to mice when administered prophylactically. Together, these data suggest that antisense PMO and P-PMO have the potential to control EBOV infection and are promising therapeutic candidates.

• Authors: Sven Enterlein, Kelly L Warfield, Dana L Swenson, David A Stein, Jeffery L Smith, C Scott Gamble, Andrew D Kroeker, Patrick L Iversen, Sina Bavari, Elke Mühlberger
• Journal: Antimicrobial agents and chemotherapy (Antimicrob Agents Chemother) Vol. 50 Issue 3 Pg. 984-93 (Mar 2006) ISSN: 0066-4804 [Print] United States
• PMID: 16495261 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, Non-P.H.S.)

Chemical References

• Morpholines
• Peptides
• RNA, Viral
• VP35 protein, filovirus
• Viral Proteins
• Viral Regulatory and Accessory Proteins

Topics

• Animals
• Chlorocebus aethiops
• Dose-Response Relationship, Drug
• Ebolavirus (drug effects, genetics)
• Female
• Hemorrhagic Fever, Ebola (immunology, mortality, prevention & control)
• Kinetics
• Mice
• Mice, Inbred C57BL
• Morpholines (pharmacology)
• Peptides (chemistry)
• RNA, Viral (genetics)
• Reverse Transcriptase Polymerase Chain Reaction
• Vero Cells
• Viral Proteins (pharmacology)
• Viral Regulatory and Accessory Proteins
• Virus Replication (drug effects, genetics)