Advanced antisense therapies for postexposure protection against lethal filovirus infections

Journal name:
Nature Medicine
Volume:
16,
Pages:
991–994
Year published:
DOI:
doi:10.1038/nm.2202
Received
Accepted
Published online

Currently, no vaccines or therapeutics are licensed to counter Ebola or Marburg viruses, highly pathogenic filoviruses that are causative agents of viral hemorrhagic fever. Here we show that administration of positively charged phosphorodiamidate morpholino oligomers (PMOplus), delivered by various dosing strategies initiated 30–60 min after infection, protects >60% of rhesus monkeys against lethal Zaire Ebola virus (ZEBOV) and 100% of cynomolgus monkeys against Lake Victoria Marburg virus (MARV) infection. PMOplus may be useful for treating these and other highly pathogenic viruses in humans.

At a glance

Figures

  1. Postexposure protection of ZEBOV-infected rhesus monkeys by AVI-6002.
    Figure 1: Postexposure protection of ZEBOV-infected rhesus monkeys by AVI-6002.

    In all experiments, monkeys were challenged with approximately 1,000 plaque-forming units of ZEBOV-Kikwit by intramuscular injection, and PMOplus was administered in PBS beginning 30–60 min after challenge. (a) Combined Kaplan-Meier survival analysis from two proof-of-concept experiments in which monkeys (n = 8) were treated daily with 40 mg per kg body weight AVI-6002. Doses were divided into equal volumes and administered at intraperitoneal and subcutaneous sites. Four monkeys received treatments for 14 d, and four received treatment for 10 d. A single untreated monkey served as an infection control. (bf) Multiple-dose postexposure efficacy assessment of AVI-6002 for treatment of ZEBOV infection in rhesus monkeys. Monkeys were randomly assigned to treatment groups, and the in-life portion of the experiment was conducted under single-blind experimental conditions. AVI-6002 was delivered at one of four dose levels: 40 (6002-40; n = 5), 28 (6002-28; n = 5), 16 (6002-16; n = 5) or 4 (6002-4; n = 5) mg per kg body weight. Four monkeys were treated with 40 mg per kg body weight negative-control PMOplus formulation (AVI-6003; 6003-40), and one monkey was treated with PBS. All treatments were administered by bolus intravenous injection daily through day 14 after infection. Statistically significant differences (*P < 0.05) between means of AVI-6002 treatments and the AVI-6003 treatment are indicated. (b) Kaplan-Meier survival curves. (c) Mean platelet counts. (d) Mean peripheral blood lymphocyte counts. (e) Mean aspartate aminotransferase (AST) concentration (instrument interference prevented lymphocyte quantification of day 28 6002-16 sample). (f) Plasma viremia assessed by quantitative real-time PCR; results from samples collected from individual monkeys on day 8 are shown.

  2. Postexposure protection of MARV-infected cynomolgus monkeys by AVI-6003.
    Figure 2: Postexposure protection of MARV-infected cynomolgus monkeys by AVI-6003.

    In all experiments, monkeys were challenged with approximately 1,000 plaque-forming units of MARV-Musoke by subcutaneous injection, and treatments were initiated beginning 30–60 min after challenge and were administered daily through day 14 after infection. PMOplus was formulated in PBS for delivery. (a) Combined survival and viremia from two independent proof-of-concept evaluations. AVI-6003 was administered at doses of 40 (n = 4) or 30 (n = 3) mg per kg body weight via intraperitoneal and subcutaneous injections or was delivered at 40 mg per kg body weight by subcutaneous (n = 3) or intravenous (n = 3) injection. Treatments delivered at intraperitoneal and subcutaneous sites were administered by injecting equal volumes at each site. Viremia results from day 8, obtained by standard plaque assay (Vero cells), are presented and are depicted as the range and geometric mean. PFU, plaque-forming units. (bf) Multiple-dose assessment of AVI-6003 for treatment of MARV infection after exposure in cynomolgus monkeys. In-life study components were conducted under single-blind experimental conditions, and monkeys were randomized to treatments. AVI-6003 was delivered intravenously at one of three doses: 30 (6003-30; n = 5), 15 (6003-15; n = 5) or 7.5 (6003-7.5; n = 5) mg per kg body weight. Four monkeys were treated with 30 mg per kg body weight negative-control PMOplus formulation (AVI-6002; 6002-30), and one monkey was treated with PBS. Statistically significant differences (*P < 0.05) between means of AVI-6003 treatments and the AVI-6002 treatment are indicated. (b) Kaplan-Meier survival curves. (c) Mean platelet counts. (d) Mean peripheral blood lymphocyte counts. (e) Mean aspartate aminotransferase concentration. (f) Plasma viremia assessed by standard plaque assay; the maximum viremia value (occurring at either at day 8 or day 10 after infection in all monkeys) obtained during the course of infection is shown for each monkey. This research was conducted in compliance with the US Animal Welfare Act and other federal statutes and regulations relating to animals and experiments involving animals, and it adheres to principles stated in the Guide for the Care and Use of Laboratory Animals, National Research Council. The studies were approved by the USAMRIID Institutional Animal Care and Use Committee, and USAMRIID is fully accredited by the Association for Assessment and Accreditation of Laboratory Animal Care International.

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Author information

  1. These authors contributed equally to this work.

    • Travis K Warren &
    • Kelly L Warfield

Affiliations

  1. United States Army Medical Research Institute of Infectious Diseases (USAMRIID), Fort Detrick, Maryland, USA.

    • Travis K Warren,
    • Kelly L Warfield,
    • Jay Wells,
    • Dana L Swenson,
    • Kelly S Donner,
    • Sean A Van Tongeren,
    • Nicole L Garza,
    • Lian Dong,
    • Donald K Nichols &
    • Sina Bavari
  2. AVI Biopharma, Corvallis, Oregon, USA.

    • Dan V Mourich,
    • Stacy Crumley &
    • Patrick L Iversen
  3. Current address: Integrated BioTherapeutics, Germantown, Maryland, USA.

    • Kelly L Warfield &
    • Dana L Swenson

Contributions

T.K.W. designed and supervised multiple-dose primate evaluations, evaluated results and wrote the manuscript. K.L.W. designed, supervised and conducted proof-of-concept nonhuman primate and rodent investigations and evaluated results. J.W., D.L.S., K.S.D., S.A.V.T. and N.L.G. conducted the nonhuman primate and rodent studies and analyzed samples. L.D. conducted quantitative PCR analysis. D.K.N. conducted post-mortem analyses of all nonhuman primate subjects. D.V.M. and S.C. were responsible for synthesis of PMOplus agents. P.L.I. and S.B. designed experiments, evaluated results and provided project oversight. All authors read and approved the final version of the manuscript.

Competing financial interests

P.L.I. and S.B. claim intellectual property regaring PMOplus technologies for treatment of viral infections. P.L.I., D.V.M. and S.C. are employees of AVI BioPharma.

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