Gene-Specific Countermeasures against Ebola Virus Based on Antisense Phosphorodiamidate Morpholino Oligomers
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{"title"=>"Gene-specific countermeasures against Ebola virus based on antisense phosphorodiamidate morpholino oligomers", "type"=>"journal", "authors"=>[{"first_name"=>"Kelly L.", "last_name"=>"Warfield", "scopus_author_id"=>"6602864764"}, {"first_name"=>"Dana L.", "last_name"=>"Swenson", "scopus_author_id"=>"7102254368"}, {"first_name"=>"Gene G.", "last_name"=>"Olinger", "scopus_author_id"=>"7006751100"}, {"first_name"=>"Donald K.", "last_name"=>"Nichols", "scopus_author_id"=>"56256393600"}, {"first_name"=>"William D.", "last_name"=>"Pratt", "scopus_author_id"=>"7201577658"}, {"first_name"=>"Robert", "last_name"=>"Blouch", "scopus_author_id"=>"12808233100"}, {"first_name"=>"David A.", "last_name"=>"Stein", "scopus_author_id"=>"7401615537"}, {"first_name"=>"M. Javad", "last_name"=>"Aman", "scopus_author_id"=>"56744332200"}, {"first_name"=>"Patrick L.", "last_name"=>"Iversen", "scopus_author_id"=>"7102956876"}, {"first_name"=>"Sina", "last_name"=>"Bavari", "scopus_author_id"=>"7003364755"}], "year"=>2006, "source"=>"PLoS Pathogens", "identifiers"=>{"sgr"=>"33644653708", "doi"=>"10.1371/journal.ppat.0020001", "issn"=>"15537366", "pui"=>"43555743", "isbn"=>"1553-7366 (Print)", "pmid"=>"16415982", "scopus"=>"2-s2.0-33644653708"}, "id"=>"dd4b07c6-41f2-3ab3-912b-4b2ec4e54fe8", "abstract"=>"The filoviruses Marburg virus and Ebola virus (EBOV) quickly outpace host immune responses and cause hemorrhagic fever, resulting in case fatality rates as high as 90% in humans and nearly 100% in nonhuman primates. The development of an effective therapeutic for EBOV is a daunting public health challenge and is hampered by a paucity of knowledge regarding filovirus pathogenesis. This report describes a successful strategy for interfering with EBOV infection using antisense phosphorodiamidate morpholino oligomers (PMOs). A combination of EBOV-specific PMOs targeting sequences of viral mRNAs for the viral proteins (VPs) VP24, VP35, and RNA polymerase L protected rodents in both pre- and post-exposure therapeutic regimens. In a prophylactic proof-of-principal trial, the PMOs also protected 75% of rhesus macaques from lethal EBOV infection. The work described here may contribute to development of designer, \"druggable\" countermeasures for filoviruses and other microbial pathogens.", "link"=>"http://www.mendeley.com/research/genespecific-countermeasures-against-ebola-virus-based-antisense-phosphorodiamidate-morpholino-oligo", "reader_count"=>65, "reader_count_by_academic_status"=>{"Unspecified"=>3, "Librarian"=>6, "Researcher"=>19, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>9, "Student > Postgraduate"=>4, "Other"=>6, "Student > Bachelor"=>9, "Lecturer > Senior Lecturer"=>1, "Professor"=>6}, "reader_count_by_user_role"=>{"Unspecified"=>3, "Librarian"=>6, "Researcher"=>19, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>9, "Student > Postgraduate"=>4, "Other"=>6, "Student > Bachelor"=>9, "Lecturer > Senior Lecturer"=>1, "Professor"=>6}, "reader_count_by_subject_area"=>{"Unspecified"=>3, "Agricultural and Biological Sciences"=>28, "Arts and Humanities"=>1, "Veterinary Science and Veterinary Medicine"=>1, "Business, Management and Accounting"=>1, "Chemistry"=>3, "Computer Science"=>1, "Earth and Planetary Sciences"=>1, "Environmental Science"=>1, "Nursing and Health Professions"=>3, "Biochemistry, Genetics and Molecular Biology"=>2, "Medicine and Dentistry"=>10, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Psychology"=>2, "Social Sciences"=>2, "Immunology and Microbiology"=>5}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>10}, "Social Sciences"=>{"Social Sciences"=>2}, "Psychology"=>{"Psychology"=>2}, "Unspecified"=>{"Unspecified"=>3}, "Environmental Science"=>{"Environmental Science"=>1}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}, "Arts and Humanities"=>{"Arts and Humanities"=>1}, "Chemistry"=>{"Chemistry"=>3}, "Earth and Planetary Sciences"=>{"Earth and Planetary Sciences"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>5}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>28}, "Computer Science"=>{"Computer Science"=>1}, "Business, Management and Accounting"=>{"Business, Management and Accounting"=>1}, "Nursing and Health Professions"=>{"Nursing and Health Professions"=>3}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>1}}, "reader_count_by_country"=>{"Canada"=>1, "United States"=>2, "United Kingdom"=>1, "Slovakia"=>1, "Spain"=>1}, "group_count"=>4}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/961133"], "description"=>"<div><p>(A) PMO-treated C57BL/6 mice that have previously survived EBOV infection generate EBOV-specific CD8<sup>+</sup> responses. Pooled splenocytes from three PMO-treated EBOV survivors were restimulated in vitro with EBOV-specific VP35 or NP peptides, an irrelevant Lassa NP peptide as a negative control, or PMA/ionomycin as a positive control. The stimulated cells were stained after for 4 h in culture with anti-CD44-FITC, anti-IFN-γ-PE, and anti-CD8-Cy-Chrome. The percent of CD44<sup>+</sup>, IFN-γ+ cells among CD8<sup>+</sup> lymphocytes is indicated in the upper right quadrant of each plot. These data are representative of several experiments examining the EBOV-specific T cell epitopes in PMO-treated mice after challenge.</p><p>(B) Total serum anti-EBOV antibodies were measured in surviving mice prior to or 4 wk following treatment and challenge. PMO mice were treated with the combination of PMOs 24 hand 4 h before challenge, and their antibody responses are compared with mice treated with Ebola VLPs 24 h before EBOV infection. The results are depicted as the endpoint titers of the individual mice (filled circles). The horizontal line in each column represents the geometric mean titer of the group.</p><p>(C) Mice that previously survived EBOV challenge following PMO treatment were rechallenged with 1,000 pfu of mouse-adapted Ebola virus 4 wk after the initial challenge. Results are plotted as percent survival for the PMO-treated mice (black) and naïve control mice (<i>n</i> = 10 per group).</p></div>", "links"=>[], "tags"=>["responses", "pmo-treated", "mice", "ebola"], "article_id"=>631120, "categories"=>["Virology", "Biotechnology", "Cell Biology", "Infectious Diseases", "Immunology"], "users"=>["Kelly L Warfield", "Dana L Swenson", "Gene G Olinger", "Donald K Nichols", "William D Pratt", "Robert Blouch", "David A Stein", "M. Javad Aman", "Patrick L Iversen", "Sina Bavari"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.0020001.g004", "stats"=>{"downloads"=>0, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Immune_Responses_of_PMO_Treated_Mice_Following_Survival_of_Ebola_Virus_Infection_/631120", "title"=>"Immune Responses of PMO-Treated Mice Following Survival of Ebola Virus Infection", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-22 08:55:10"}
  • {"files"=>["https://ndownloader.figshare.com/files/960989"], "description"=>"<div><p>(A–D) C57BL/6 mice were challenged intraperitoneally with 1,000 plaque-forming units of EBOV following treatment with PMOs. Immunoperoxidase stain showing presence of viral antigen is brown with hematoxylin counterstain. Spleen (100×) of a mouse treated with scrambled PMO (A) or the EBOV VP24, VP35, and L PMOs (B) 3 d after EBOV infection. Diffuse staining pattern in the livers (600×) of the scrambled PMO-treated mice (C) on day 6 of EBOV infection, compared with focal areas of infection in the mice treated with the combination of PMOs (D).</p><p>(E) Viral titers in tissues from mice treated with a combination of 3 PMO and infected with 1,000 pfu of EBOV. Samples of the liver, spleen, and kidney were taken at 3 d or 6 d post challenge (dpc), macerated, and analyzed for viral titer using plaque assay. The data are presented as the mean viral titer of three mice with errors bars representing the standard deviation.</p></div>", "links"=>[], "tags"=>["slows", "viral", "replication"], "article_id"=>630983, "categories"=>["Virology", "Biotechnology", "Cell Biology", "Infectious Diseases", "Immunology"], "users"=>["Kelly L Warfield", "Dana L Swenson", "Gene G Olinger", "Donald K Nichols", "William D Pratt", "Robert Blouch", "David A Stein", "M. Javad Aman", "Patrick L Iversen", "Sina Bavari"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.0020001.g003", "stats"=>{"downloads"=>1, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_PMO_Treatment_Slows_Viral_Replication_in_Mice_/630983", "title"=>"PMO Treatment Slows Viral Replication in Mice", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-22 08:54:09"}
  • {"files"=>["https://ndownloader.figshare.com/files/961291"], "description"=>"<div><p>(A) Treatment schedule for proof-of-concept trial using PMO to treat lethal Ebola virus infection in nonhuman primates.</p><p>(B) Survival following infection with 1,000 pfu of EBOV in monkeys treated with a combination of PMOs (▪, <i>n</i> = 4) or untreated monkeys (○, <i>n</i> = 3). The arrows indicate the time of death of each PMO-treated monkey.</p><p>(C–E) Viral titers (C), platelet counts (D), or alkaline phosphatase levels (E) in the blood of the PMO-treated monkeys [0646 (♦), 1438 (▴), 1496 (×), 1510 (▪)] or a representative untreated monkey (○).</p></div>", "links"=>[], "tags"=>["rhesus", "macaques", "antisense", "pmos", "lethal", "ebola"], "article_id"=>631267, "categories"=>["Virology", "Biotechnology", "Cell Biology", "Infectious Diseases", "Immunology"], "users"=>["Kelly L Warfield", "Dana L Swenson", "Gene G Olinger", "Donald K Nichols", "William D Pratt", "Robert Blouch", "David A Stein", "M. Javad Aman", "Patrick L Iversen", "Sina Bavari"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.0020001.g006", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Treatment_of_Rhesus_Macaques_with_Antisense_PMOs_Provide_Protection_against_Lethal_Ebola_Virus_Infection_/631267", "title"=>"Treatment of Rhesus Macaques with Antisense PMOs Provide Protection against Lethal Ebola Virus Infection", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-22 08:56:13"}
  • {"files"=>["https://ndownloader.figshare.com/files/960839"], "description"=>"<div><p>(A–C) Survival of mice (<i>n</i> = 10) pretreated at 4 h and 24 h before EBOV infection with 500 (square) or 50 (triangle) μg doses of PMOs targeting EBOV- (filled) or MARV-specific (unfilled) VP24 (A), VP35 (B), L (C), or with PBS (×).</p><p>(D) Survival of mice (<i>n</i> = 10) pretreated at 24 h and 4 h before EBOV infection with 500 μg (square), 50 μg (triangle), or 5 μg (circle) doses of a combination of the EBOV- (filled) or MARV-specific (unfilled) VP24, VP35, and L PMOs or PBS (×).</p><p>(E) Survival of mice (<i>n</i> = 10) treated 4 h before (unfilled) or 24 h after (filled) EBOV infection with 500 μg (square), 50 μg (triangle), or 5 μg (circle) of the combination of EBOV-specific PMOs or an unrelated sequence (×). All the animals in these studies were monitored for at least 28 d for illness, and there were no differences observed in survival within any of the groups between 14 d and 28 d.</p></div>", "links"=>[], "tags"=>["pmos", "mice", "lethal", "ebola"], "article_id"=>630832, "categories"=>["Virology", "Biotechnology", "Cell Biology", "Infectious Diseases", "Immunology"], "users"=>["Kelly L Warfield", "Dana L Swenson", "Gene G Olinger", "Donald K Nichols", "William D Pratt", "Robert Blouch", "David A Stein", "M. Javad Aman", "Patrick L Iversen", "Sina Bavari"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.0020001.g002", "stats"=>{"downloads"=>1, "page_views"=>15, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Ebola_Specific_PMOs_Protect_Mice_from_Lethal_Ebola_Virus_Infection_/630832", "title"=>"Ebola-Specific PMOs Protect Mice from Lethal Ebola Virus Infection", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-22 08:53:06"}
  • {"files"=>["https://ndownloader.figshare.com/files/961214"], "description"=>"<div><p>Hartley guinea pigs (<i>n</i> = 6) weighing 400–500 g each were treated intraperitoneally with 10 mg each of VP24, VP35, and L PMO in PBS at −24 (triangle), +24 (circle), or +96 (square) h post challenge. Control guinea pigs were injected with PBS only (diamond). The guinea pigs were infected subcutaneously with ~1,000 pfu of EBOV and monitored for illness for 21 d.</p><p>(A) The data are presented as percent survival for each group.</p><p>(B) Viral titers in the serum of guinea pigs at 7 d post challenge (dpc). The data are presented as the titers of individual guinea pigs treated with PMOs at −24, +24, or +96 h post challenge or PBS only and the mean viral titer for each group (horizontal line). Animals which survived are represented by open symbols and those that succumbed to disease are represented by filled symbols.</p></div>", "links"=>[], "tags"=>["guinea", "pigs", "antisense", "pmos", "lethal", "ebola"], "article_id"=>631196, "categories"=>["Virology", "Biotechnology", "Cell Biology", "Infectious Diseases", "Immunology"], "users"=>["Kelly L Warfield", "Dana L Swenson", "Gene G Olinger", "Donald K Nichols", "William D Pratt", "Robert Blouch", "David A Stein", "M. Javad Aman", "Patrick L Iversen", "Sina Bavari"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.0020001.g005", "stats"=>{"downloads"=>0, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Treatment_of_Guinea_Pigs_with_Antisense_PMOs_Increases_Survival_Following_Lethal_Ebola_Virus_Infection_/631196", "title"=>"Treatment of Guinea Pigs with Antisense PMOs Increases Survival Following Lethal Ebola Virus Infection", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-22 08:55:44"}
  • {"files"=>["https://ndownloader.figshare.com/files/960752"], "description"=>"<div><p>(A) Schematic diagram of the EBOV genome showing the PMO sequences and the relative locations of their mRNA targets.</p><p>(B–D) Sequence-specific inhibition of EBOV VP24 (B), VP35 (C), and L (D) gene targets in cell-free translation assay. Plasmids containing 150 nucleotides encompassing the PMO target area from EBOV VP24, VP35, or L fused to firefly luciferase were used to generate RNA. Inhibition of the VP24, VP35, or L RNA targets was assessed using in vitro translation reactions containing different concentrations of PMO targeting EBOV (•) or MARV (▪) VP24, VP35, or L, respectively, or a scrambled (▴) PMO, along with the respective RNA. The data are shown as means of three replicates per data point and the standard error of the mean.</p><p>(E) Inhibition of viral amplification by PMOs in vitro. Vero E6 cells were pretreated for 24 h with 20 μM of EBOV VP24-specific PMO (blue), EBOV VP35-specific PMO (red), EBOV L-specific PMO (yellow), combination of the three EBOV-specific PMOs (gray), or a scrambled PMO (black). The cells were infected with EBOV-Zaire (multiplicity of infection = 1), and the viral titers were assessed at 24, 48, and 72 h post infection. The data are presented as percent reduction of EBOV-specific PMO-treated cells, as compared with the scrambled PMO-treated cells (titers at 24 h were 127 pfu/ml, at 48 h were 4,241 pfu/ml, and at 72 h were 202,000 pfu/ml). The experiments were performed at least three times and a single representative experiment is shown.</p></div>", "links"=>[], "tags"=>["pmos", "inhibit", "ebola"], "article_id"=>630742, "categories"=>["Virology", "Biotechnology", "Cell Biology", "Infectious Diseases", "Immunology"], "users"=>["Kelly L Warfield", "Dana L Swenson", "Gene G Olinger", "Donald K Nichols", "William D Pratt", "Robert Blouch", "David A Stein", "M. Javad Aman", "Patrick L Iversen", "Sina Bavari"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.0020001.g001", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Gene_Specific_PMOs_Inhibit_Ebola_Virus_Gene_Translation_and_Growth_/630742", "title"=>"Gene-Specific PMOs Inhibit Ebola Virus Gene Translation and Growth", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-22 08:52:22"}

PMC Usage Stats | Further Information

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Relative Metric

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