VP35 knockdown inhibits Ebola virus amplification and protects against lethal infection in mice.
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.
Department of Virology, Philipps-University Marburg, Hans-Meerwein-Str. 3, 35043 Marburg, Germany.
Sven
S
Enterlein
Kelly L
KL
Warfield
Dana L
DL
Swenson
David A
DA
Stein
Jeffery L
JL
Smith
C Scott
CS
Gamble
Andrew D
AD
Kroeker
Patrick L
PL
Iversen
Sina
S
Bavari
Elke
E
Mühlberger
D000818:Animals; D002522:Chlorocebus aethiops; D004305:Dose-Response Relationship, Drug; D029043:Ebolavirus / Q000187:drug effects* / Q000235:genetics; D005260:Female; D019142:Hemorrhagic Fever, Ebola / Q000276:immunology / Q000401:mortality* / Q000517:prevention & control*; D007700:Kinetics; D051379:Mice; D008810:Mice, Inbred C57BL; D009025:Morpholines / Q000494:pharmacology*; D010455:Peptides / Q000737:chemistry; D012367:RNA, Viral / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014709:Vero Cells; D014764:Viral Proteins / Q000494:pharmacology*; D054334:Viral Regulatory and Accessory Proteins; D014779:Virus Replication / Q000187:drug effects / Q000235:genetics
D009025:Morpholines; D010455:Peptides; D012367:RNA, Viral; C080980:VP35 protein, filovirus; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins