Antiviral and Neuroprotective Role of Octaguanidinium Dendrimer-Conjugated Morpholino Oligomers in Japanese Encephalitis
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{"title"=>"Antiviral and neuroprotective role of octaguanidinium dendrimer-conjugated Morpholino oligomers in Japanese encephalitis", "type"=>"journal", "authors"=>[{"first_name"=>"Arshed", "last_name"=>"Nazmi", "scopus_author_id"=>"35386900600"}, {"first_name"=>"Kallol", "last_name"=>"Dutta", "scopus_author_id"=>"30067532400"}, {"first_name"=>"Anirban", "last_name"=>"Basu", "scopus_author_id"=>"35335904100"}], "year"=>2010, "source"=>"PLoS Neglected Tropical Diseases", "identifiers"=>{"pmid"=>"21124882", "sgr"=>"78649770754", "doi"=>"10.1371/journal.pntd.0000892", "scopus"=>"2-s2.0-78649770754", "pui"=>"360094647", "isbn"=>"1935-2735 (Electronic)\\r1935-2727 (Linking)", "issn"=>"1935-2735"}, "id"=>"0a0142ae-61af-30cf-a3d2-612a176a0d1f", "abstract"=>"BACKGROUND: Japanese encephalitis (JE), caused by a mosquito-borne flavivirus, is endemic to the entire south-east Asian and adjoining regions. Currently no therapeutic interventions are available for JE, thereby making it one of the most dreaded encephalitides in the world. An effective way to counter the virus would be to inhibit viral replication by using anti-sense molecules directed against the viral genome. Octaguanidinium dendrimer-conjugated Morpholino (or Vivo-Morpholino) are uncharged anti-sense oligomers that can enter cells of living organisms by endocytosis and subsequently escape from endosomes into the cytosol/nuclear compartment of cells. We hypothesize that Vivo-Morpholinos generated against specific regions of 3' or 5' untranslated regions of JEV genome, when administered in an experimental model of JE, will have significant antiviral and neuroprotective effect.\\n\\nMETHODOLOGY/PRINCIPAL FINDINGS: Mice were infected with JEV (GP78 strain) followed by intraperitoneal administration of Morpholinos (5 mg/kg body weight) daily for up to five treatments. Survivability of the animals was monitored for 15 days (or until death) following which they were sacrificed and their brains were processed either for immunohistochemical staining or protein extraction. Plaque assay and immunoblot analysis performed from brain homogenates showed reduced viral load and viral protein expression, resulting in greater survival of infected animals. Neuroprotective effect was observed by thionin staining of brain sections. Cytokine bead array showed reduction in the levels of proinflammatory cytokines in brain following Morpholino treatment, which were elevated after infection. This corresponded to reduced microglial activation in brain. Oxidative stress was reduced and certain stress-related signaling molecules were found to be positively modulated following Morpholino treatment. In vitro studies also showed that there was decrease in infective viral particle production following Morpholino treatment.\\n\\nCONCLUSIONS/SIGNIFICANCE: Administration of Vivo-Morpholino effectively resulted in increased survival of animals and neuroprotection in a murine model of JE. Hence, these oligomers represent a potential antiviral agent that merits further evaluation.", "link"=>"http://www.mendeley.com/research/antiviral-neuroprotective-role-octaguanidinium-dendrimerconjugated-morpholino-oligomers-japanese-enc", "reader_count"=>25, "reader_count_by_academic_status"=>{"Unspecified"=>3, "Librarian"=>1, "Researcher"=>9, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>3, "Student > Postgraduate"=>1, "Student > Master"=>1, "Other"=>3, "Lecturer"=>1, "Professor"=>2}, "reader_count_by_user_role"=>{"Unspecified"=>3, "Librarian"=>1, "Researcher"=>9, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>3, "Student > Postgraduate"=>1, "Student > Master"=>1, "Other"=>3, "Lecturer"=>1, "Professor"=>2}, "reader_count_by_subject_area"=>{"Unspecified"=>4, "Environmental Science"=>1, "Biochemistry, Genetics and Molecular Biology"=>1, "Agricultural and Biological Sciences"=>14, "Neuroscience"=>3, "Veterinary Science and Veterinary Medicine"=>2}, "reader_count_by_subdiscipline"=>{"Neuroscience"=>{"Neuroscience"=>3}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>14}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>1}, "Unspecified"=>{"Unspecified"=>4}, "Environmental Science"=>{"Environmental Science"=>1}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>2}}, "reader_count_by_country"=>{"United Kingdom"=>1, "India"=>3, "French Polynesia"=>1}, "group_count"=>1}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/407165"], "description"=>"<div><h3>Background</h3><p>Japanese encephalitis (JE), caused by a mosquito-borne flavivirus, is endemic to the entire south-east Asian and adjoining regions. Currently no therapeutic interventions are available for JE, thereby making it one of the most dreaded encephalitides in the world. An effective way to counter the virus would be to inhibit viral replication by using anti-sense molecules directed against the viral genome. Octaguanidinium dendrimer-conjugated Morpholino (or Vivo-Morpholino) are uncharged anti-sense oligomers that can enter cells of living organisms by endocytosis and subsequently escape from endosomes into the cytosol/nuclear compartment of cells. We hypothesize that Vivo-Morpholinos generated against specific regions of 3′ or 5′ untranslated regions of JEV genome, when administered in an experimental model of JE, will have significant antiviral and neuroprotective effect.</p><h3>Methodology/Principal Findings</h3><p>Mice were infected with JEV (GP78 strain) followed by intraperitoneal administration of Morpholinos (5 mg/kg body weight) daily for up to five treatments. Survivability of the animals was monitored for 15 days (or until death) following which they were sacrificed and their brains were processed either for immunohistochemical staining or protein extraction. Plaque assay and immunoblot analysis performed from brain homogenates showed reduced viral load and viral protein expression, resulting in greater survival of infected animals. Neuroprotective effect was observed by thionin staining of brain sections. Cytokine bead array showed reduction in the levels of proinflammatory cytokines in brain following Morpholino treatment, which were elevated after infection. This corresponded to reduced microglial activation in brain. Oxidative stress was reduced and certain stress-related signaling molecules were found to be positively modulated following Morpholino treatment. <em>In vitro</em> studies also showed that there was decrease in infective viral particle production following Morpholino treatment.</p><h3>Conclusions/Significance</h3><p>Administration of Vivo-Morpholino effectively resulted in increased survival of animals and neuroprotection in a murine model of JE. Hence, these oligomers represent a potential antiviral agent that merits further evaluation.</p></div>", "links"=>[], "tags"=>["antiviral", "neuroprotective", "octaguanidinium", "dendrimer-conjugated", "morpholino", "oligomers", "japanese", "encephalitis"], "article_id"=>140448, "categories"=>["Cancer", "Medicine"], "users"=>["Arshed Nazmi", "Kallol Dutta", "Anirban Basu"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0000892", "stats"=>{"downloads"=>1, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Antiviral_and_Neuroprotective_Role_of_Octaguanidinium_Dendrimer_Conjugated_Morpholino_Oligomers_in_Japanese_Encephalitis/140448", "title"=>"Antiviral and Neuroprotective Role of Octaguanidinium Dendrimer-Conjugated Morpholino Oligomers in Japanese Encephalitis", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-11-23 00:07:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/816996"], "description"=>"<p>Approximately 4-fold increases in the levels of pNFκB and 3-fold increases in the levels of phosphoP38MAPK in JEV-infected and JEV+SC-MO groups were observed in comparison to Sham. The levels of both pNFκB and phosphoP38MAPK were significantly reduced in JEV+3′ MO and JEV+5′ MO groups when compared to only JEV-infected groups (A–C). Phospho ERK1 and ERK2 levels were found to be increased by approximately 3- and 5-fold in JEV-infected and JEV+SC-MO groups respectively, when compared to Sham. but showed considerable decreases in JEV+3′ MO and JEV+5′ MO groups when compared to only JEV-infected groups (A&D) (<sup>*</sup> p<0.01 for JEV-infected and JEV+SC-MO when compared to Sham; <sup>#</sup> p<0.01 for JEV+3′ MO and JEV+5′MO when compared to only JEV-infected groups).</p>", "links"=>[], "tags"=>["regulates", "proteins"], "article_id"=>487369, "categories"=>["Infectious Diseases", "Medicine"], "users"=>["Arshed Nazmi", "Kallol Dutta", "Anirban Basu"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0000892.g006", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_MOs_regulates_the_expression_pattern_of_several_key_proteins_associated_with_stress_/487369", "title"=>"MOs regulates the expression pattern of several key proteins associated with stress.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-11-23 02:02:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/816873"], "description"=>"<p>Increases were observed in the ROS levels in the brain samples of JEV-infected and JEV+SC-MO groups in comparison to Sham, that were reduced following 3′ and 5′ MO treatments. Although ROS levels were decreased in JEV+3′ MO group when compared to JEV-infected group, it remained significantly higher than that in Sham (A). Approximately 13-fold increases were observed in the levels of HSP-70 in JEV-infected and JEV+SC-MO groups as compared to Sham. These drastic increases were significantly reduced in JEV+3′ MO and JEV+5′ MO groups, the levels remained significantly higher than Sham (B&C). SOD-1 levels were found to be elevated 2- and nearly 3-fold in JEV-infected and JEV+SC-MO groups respectively compared to Sham. 3′ and 5′ MO treatment caused significant reduction of SOD-1 levels compared to JEV-infected group (B&D). Alterations in TRX-1 levels were similar to that observed in SOD-1 except that its level in JEV+ SC-MO was not significantly different than only JEV-infected group (B&E). Nearly 2-fold increases were observed in NO levels of JEV-infected and JEV+SC-MO groups when compared to those obtained from Sham. NO levels were subsequently reduced to significantly lower levels following 3′ and 5′MO treatments (F). iNOS expression was found to increase 8-fold in JEV-infected and JEV+SC-MO groups when compared to Sham. Following 3′ and 5′MO treatments iNOS levels decreased significantly as compared to JEV-infected group (G&H). (<sup>*</sup> p<0.01 for JEV when compared to Sham; <sup>**</sup> p<0.01 for JEV+SC-MO when compared to JEV-infected only;<sup> #</sup> p<0.01 for JEV+ 3′MO and JEV+ 5′ MO when compared to only JEV-infected group; ̂ p<0.01 for JEV+ 3′MO when compared to Sham; <sup>$</sup> p<0.01 for JEV+ 5′ MO when compared to Sham).</p>", "links"=>[], "tags"=>["modulates", "oxidative", "markers"], "article_id"=>487240, "categories"=>["Infectious Diseases", "Medicine"], "users"=>["Arshed Nazmi", "Kallol Dutta", "Anirban Basu"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0000892.g005", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_MOs_treatment_modulates_oxidative_stress_markers_in_CNS_/487240", "title"=>"MOs treatment modulates oxidative stress markers in CNS.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-11-23 02:00:40"}
  • {"files"=>["https://ndownloader.figshare.com/files/816422"], "description"=>"<p>Schematic diagram showing the secondary structure of the 3′ and 5′ UTR of JEV genomic RNA as predicted by the Mfold program <a href=\"http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0000892#pntd.0000892-Zuker1\" target=\"_blank\">[41]</a>. The regions blocked by the 3′ MO and 5′ MO are marked by black curved lines and marked with arrows. Numbers in the smaller font refer to the nucleotide position of the JEV genome.</p>", "links"=>[], "tags"=>["jev", "genome", "blocked"], "article_id"=>486790, "categories"=>["Infectious Diseases", "Medicine"], "users"=>["Arshed Nazmi", "Kallol Dutta", "Anirban Basu"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0000892.g001", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Regions_of_the_JEV_GP78_strain_genome_that_are_blocked_by_the_MOs_/486790", "title"=>"Regions of the JEV (GP78 strain) genome that are blocked by the MOs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-11-23 01:53:10"}
  • {"files"=>["https://ndownloader.figshare.com/files/816606"], "description"=>"<p>Significant reduction in viral titer was observed following MO treatment in animals, as compared to JEV-infected and JEV+SC-MO groups. (<sup>*</sup> p<0.001 for JEV and JEV+SC-MO when compared to Sham, <sup>#</sup> p<0.001 for JEV+ 3′MO and JEV+ 5′ MO when compared to only JEV-infected group) (<i>A</i>). Immunoblot analysis showed expression of JEV NS5 and E glycoprotein were significantly increased in JEV-infected and JEV+SC-MO than Sham, but reduced significantly after both 3′ and 5′ MO treatments (<sup>*</sup> p<0.01 for JEV and JEV+SC-MO when compared to Sham, <sup>#</sup> p<0.01 for JEV+ 3′MO and JEV+ 5′ MO when compared to only JEV-infected group) (<i>B–D</i>). Immunostaining of brain sections from different treatment groups showed greater presence of JEV antigen in JEV and JEV+SC-MO groups (<i>E</i>). Magnification ×20; scale bar correspond to 50µ Photomicrographs shown here in this figure are representative of three individual animals from each group.</p>", "links"=>[], "tags"=>["viral"], "article_id"=>486977, "categories"=>["Infectious Diseases", "Medicine"], "users"=>["Arshed Nazmi", "Kallol Dutta", "Anirban Basu"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0000892.g003", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_MOs_treatment_reduces_the_viral_load_in_vivo_/486977", "title"=>"MOs treatment reduces the viral load <i>in vivo</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-11-23 01:56:17"}
  • {"files"=>["https://ndownloader.figshare.com/files/817133"], "description"=>"<p>NA- Not applicable.</p>", "links"=>[], "tags"=>["sequences", "locations"], "article_id"=>487504, "categories"=>["Infectious Diseases", "Medicine"], "users"=>["Arshed Nazmi", "Kallol Dutta", "Anirban Basu"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0000892.t001", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Names_sequences_and_target_locations_of_used_Vivo_Morpholinos_/487504", "title"=>"Names, sequences and target locations of used Vivo-Morpholinos.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2010-11-23 02:05:04"}
  • {"files"=>["https://ndownloader.figshare.com/files/816500"], "description"=>"<p>The survival of mice following JEV infection was dramatically increased with treatments of both 3′ and 5′ MO, though the survival in 3′ MO treated mice was greater (∼90%) than those treated with 5′MO (75%) (A). Considerable changes in the average body weights of JEV-infected animals treated with both 3′ and 5′ MO were not observed when compared to animals belonging to JEV and JEV+ SC-MO groups that showed significant reductions in their body weights from 6<sup>th</sup> day post infection till their death. Black arrows points to the days by which all the animals died. (B). Infection with JEV was accompanied with distinct symptoms that were alleviated following treatments with both 3′ and 5′ MO. Animals were assigned scores according to the symptoms, in a blinded manner. The graph was plotted by taking the scores of one animal that was considered as the representative of that group (C). n = 8 for all experiments; data shown are representative of duplicate sets of experiments.</p>", "links"=>[], "tags"=>["protected", "jev", "mo"], "article_id"=>486870, "categories"=>["Infectious Diseases", "Medicine"], "users"=>["Arshed Nazmi", "Kallol Dutta", "Anirban Basu"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0000892.g002", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Mice_are_protected_from_JEV_following_MO_treatment_/486870", "title"=>"Mice are protected from JEV following MO treatment.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-11-23 01:54:30"}
  • {"files"=>["https://ndownloader.figshare.com/files/816740"], "description"=>"<p>Thionin staining of brain sections from all the treatment groups showed neurons with distinct morphology in Sham-treated, JEV+3′ MO and JEV+5′ MO groups. However in sections from JEV-infected and JEV+SC-MO groups showed damaged neurons with altered morphology. Magnification ×20; scale bar correspond to 50 µ. Immunofluorescent staining for microglia-specific Iba-1 performed in brain sections of all groups showed that number of activated (star shaped) microglia appeared to be more frequent in JEV-infected and JEV+SC-MO groups as compared to compared to sections belonging to Sham, JEV+3′ MO and JEV+5′ MO groups. Magnification ×20; scale bar correspond to 50 µ (B). Photomicrographs shown here in this figure are representative of three individual animals from each group. CBA showed levels of MCP-1, IFN-γ, TNF-α, and IL-6 were increased significantly in both JEV-infected and JEV+SC-MO groups when compared to Sham treated groups. The elevated levels of these proinflammatory cytokines were then significantly reduced with 3′ and 5′ MO treatments (<sup>*</sup> p<0.01 for JEV and JEV+SC-MO when compared to Sham; <sup>#</sup> p<0.01 for JEV+ 3′MO and JEV+ 5′ MO when compared to only JEV-infected group) (C–F).</p>", "links"=>[], "tags"=>["microglial", "activation", "inhibits", "proinflammatory", "cytokines"], "article_id"=>487110, "categories"=>["Infectious Diseases", "Medicine"], "users"=>["Arshed Nazmi", "Kallol Dutta", "Anirban Basu"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0000892.g004", "stats"=>{"downloads"=>1, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_MOs_neuroprotect_reduces_microglial_activation_and_inhibits_proinflammatory_cytokines_production_in_brain_/487110", "title"=>"MOs neuroprotect, reduces microglial activation and inhibits proinflammatory cytokines production in brain.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2010-11-23 01:58:30"}

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