Development of therapeutic approach for progressive multifocal leukoencephalopathy using siRNA against JCV Agnogene.

2006 Fiscal Year Final Research Report Summary

• Project/Area Number: 15300112
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Nerve anatomy/Neuropathology
• Research Institution: Hokkaido University
• Principal Investigator: SAWA Hirofumi Hokkaido University, Research Center for Zoonosis Control, Dept. of Molecular Pathobiology, Professor, 人獣共通感染症リサーチセンター, 教授 (30292006)
• Co-Investigator(Kenkyū-buntansha): TAKAHASHI Hidehiro National Institute of Infectious Diseases, Department of Pathology, Section Chief, 感染病理部, 室長 (70260271) ; MOTOYAMA Noboru National Institute for Longevity Sciences, National Center for Geriatrics and Gerontology, Department of Geriatric Research, Section Chief, 研究所・老年病研究部, 室長 (50277282)
• Project Period (FY): 2003 – 2006
• Keywords: JC virus / Agnoprotein / siRNA

Research Abstract

In this project, we have attempted to inhibit JC virus (JCV) infection using small interfering RNAs (siRNAs) against JCV agnoprotein (Agno) as therapeutic strategy. In order to inhibit JCV infection in infected cells, we synthesized double-stranded siRNAs which are specific for JCV coding protein, Agno and introduced these into infected human glial-derived cells. We have observed that in post-infection treatment with siRNAs targeting JCV Agno suppresses virus production in JCV infected cells. Based on the results, we applied two patents and established the paper (J Virol 78: 7270, 2004). In addition, we have established JCV infection mimicking model using mice which were intracerebrally inoculated JCVinfected cells, and applied siRNA against JCV Agno to the model in vivo. After treatment with the siRNA, the number of intracerebral JCVinfected cells was significantly decreased. In addition, we have found that clinical drug was effectively suppressed JCV infection by modification of phosphorylation of a protein in vitro. In the future, the drug might be applicable for the JCV infected progressive multifocal leukoencephalopathy cases. We also examined the mechanism of intracellular translocation of JCV virion using the yeast two-hybrid assay using JCV Agno as a bait. We identified two cellular proteins as Agno-binding proteins. These proteins played a pivotal role in nuclear egress of JCV virion. These results were established (EMBO Rep 6: 452, 2005 and J Biol Chem 280, 24948, 2005). We also established some articles concerning about interaction between viral infection and host response.
10. KEYWORDS

Research Products

• [Journal Article] Identification of DDX1 as a JC virus (JCV) transcriptional control region-binding protein. (2007)
  • Author(s): Sunden Y, Semba S, Suzuki T, Okada Y, Orba Y, Nagashima K, Umemura T, Sawa H^*
  • Journal Title: Microbiol Immunol 51 Pages: 327-337
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] DDX1 promotes proliferation of JC virus (JCV) through transactivation of the JCV promoter. (2007)
  • Author(s): Sunden Y, Semba S, Suzuki T, Okada Y, Orba Y, Nagashima K, Umemura T, Sawa H^*
  • Journal Title: Microbiol Immunol 51 Pages: 339-347
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] JCウイルスの最近の基礎的知見。 (2007)
  • Author(s): 澤 洋文, 鈴木 忠樹, 大場 靖子, 寸田 祐嗣, 長嶋 和郎
  • Journal Title: BRAIN and NERVE 59 Pages: 101-108
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] DDX1 promotes proliferation of JC virus (JCV) through transactivation of the JCV promoter. (*corresponding author) (2007)
  • Author(s): Sunden Y, Semba S, Suzuki T, Okada Y, Orba Y, Nagashima K, Umemura T, Sawa H*
  • Journal Title: Microbiol Immunol 51 Pages: 339-347
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Identification of DDX1 as a JC virus (JCV) transcriptional control region-binding protein. (*corresponding author) (2007)
  • Author(s): Sunden Y, Semba S, Suzuki T, Okada Y, Orba Y, Nagashima K, Umemura T, Sawa H*
  • Journal Title: Microbiol Immunol 51 Pages: 327-337
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Intranasal administration of adjuvant-combined recombinant influenza virus HA vaccine protects mice from the lethal H5N1 virus infection. (2006)
  • Author(s): Asahi-Ozaki Y, Itamura S, Ichinohe T, Strong P, Tamura S, Takahashi H, Sawa H, Moriyama M, Tashiro M, Sata T, Kurata T, Hasegawa H
  • Journal Title: Microbes Infect 8 Pages: 2706-2714
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Thymus-derived leukemia-lymphoma in mice transgenic for the Tax gene of human T-lymphotropic virus type-I. (2006)
  • Author(s): Hasegawa H^*, Sawa H^*, Lewis MJ, Orba Y, Sherhy N, Yamamoto Y, Ichinohe T, Tsunetsugu-Yokota Y, Katano H, Takahashi H, Matsuda J, Sata T, Kurata T, Nagashima, K, Hall, WW
  • Journal Title: Nat Med 12 Pages: 466-472
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Characterization and application of polyclonal antibodies that specifically recognize JC virus large Tantigen. (2006)
  • Author(s): Sunden Y, Suzuki T, Orba Y, Umemura T, Asamoto M, Nagashima K, Tanaka S, Sawa H^*
  • Journal Title: Acta Neuropathol 111 Pages: 379-387
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Dicer and positive charge of proteins decrease the stability of RNA containing the AU-rich element of GM-CSF. (2006)
  • Author(s): Takahashi H, Maeda M, Sawa H, Hasegawa H, Moriyama M, Sata T, Hall WW, Kurata T
  • Journal Title: Biochem Biophys Res Commun 340 Pages: 807-814
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Tristetraprolin inhibits HIV-1 production by binding to genomic RNA. (2006)
  • Author(s): Maeda M, Sawa H, Tobiume M, Tokunaga K, Hasegawa H, Ichinohe T, Sata T, Moriyama M, Hall WW, Kurata T, Takahashi T
  • Journal Title: Microbes Infect 8 Pages: 2647-2656
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Protection against influenza virus infection by intranasal vaccine with surf clam microparticles (SMP) as an adjuvant. (2006)
  • Author(s): Ichinohe T, Watanabe I, Tao E, Ito S, Kawaguchi A, Tamura SI, Takahashi H, Sawa H, Moriyama M, Chiba J, Komase K, Suzuki Y, Kurata T, Sata T, Hasegawa H
  • Journal Title: J Med Virol 78 Pages: 954-963
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Development of Thymus-Derived T-cell Leukemia/Lymphoma in Mice Transgenic for the Tax gene of Human T-Lymphotropic Virus Type-I (HTLV-I). (*These authors contributed equally to this work.) (2006)
  • Author(s): Hasegawa H*, Sawa H*, Lewis MJ, Orba Y, Sherhy N, Yamamoto Y, Ichinohe T, Tsunetsugu-Yokota Y, Katano H, Takahashi H, Matsuda J, Sata T, Kurata T, Nagashima, K, Hall, WW.
  • Journal Title: Nat Med 12 Pages: 466-472
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Characterization and application of polyclonal antibodies that specifically recognize JC virus large T antigen. (*corresponding author) (2006)
  • Author(s): Sunden Y, Suzuki T, Orba Y, Umemura T, Asamoto M, Nagashima K, Tanaka S, Sawa H*
  • Journal Title: Acta Neuropathol 111 Pages: 379-387
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Protection against influenza virus infection by intranasal vaccine with surf clam microparticles (SM-P) as an adjuvant. (2006)
  • Author(s): Ichinohe T, Watanabe I, Tao E, Ito S, Kawaguchi A, Tamura SI, Takahashi H, Sawa H, Moriyama M, Chiba J, Komase K, Suzuki Y, Kurata T, Sata T, Hasegawa H
  • Journal Title: J Med Virol 78 Pages: 954-963
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Identification of FEZ1 as a protein that interacts with JC virus agnoprotein and microtubules : role of agnoprotein-induced dissociation of FEZ1 from microtubules in viral propagation. (2005)
  • Author(s): Suzuki T, Okada Y, Semba S, Orba Y, Yamanouchi S, Endo S, Tanaka S, Fujita T, Kuroda S, Nagashima K, Sawa H^*
  • Journal Title: J Biol Chem 280 Pages: 24948-24956
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Dissociation of heterochromatin protein 1 from lamin B receptor induced by human polyomavirus agnoprotein : role in nuclear egress of viral particles. (2005)
  • Author(s): Okada Y, Suzuki T, Sunden Y, Orba Y, Kose S, Imamoto N, Takahashi H, Tanaka S, Hall WW, Nagashima K, Sawa H^*
  • Journal Title: EMBO Rep 6 Pages: 452-457
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] The agnoprotein of polyomaviruses : A multifunctional auxiliary protein. (2005)
  • Author(s): Khalili K, White MK, Sawa H, Nagashima K, Safak M
  • Journal Title: J Cell Physiol 204 Pages: 1-7
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Isolation of a monoclonal antibody recognizing a cell-surface molecule as a receptor for JC virus. (2005)
  • Author(s): Henmi C, Sawa H^*, Iwata H, Orba Y, Tanaka S, Nagashima K
  • Journal Title: Biochem Biophys Res Commun 327 Pages: 242-251
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Synthetic double-stranded RNA [poly (I : C)] combined with mucosal vaccine protects against influenza virus infection. (2005)
  • Author(s): Ichinohe T, Watanabe I, Ito S, Moriyama M, Tamura S, Takahashi H, Sawa H, Chiba J, Kurata T, Sata T, Hasegawa H
  • Journal Title: J Virol 79 Pages: 2910-2919
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Protection against influenza virus infection by intranasal administration of HA vaccine with chitin microparticles as an adjuvant. (2005)
  • Author(s): Hasegawa H, Ichinohe T, Strong P, Watanabe I, Ito S, Tamura S-i, Takahashi H, Sawa H, Chiba J, Kurata T, Sata T
  • Journal Title: J Med Virol 75 Pages: 130-136
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Clinicopathological and virological analyses of familial HTLV-I associated polyneuropathy. (2005)
  • Author(s): Sawa H, Nagashima T, Nagashima K, Shinohara T, Chuma T, Mano Y, Tachi N, Hall WW
  • Journal Title: J Neurovirol 11 Pages: 199-207
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Identification of FEZ1 as a protein that interacts with JC virus agnoprotein and microtubules : role of agnoprotein-induced dissociation of FEZ1 from microtubules in viral propagation. (* corresponding author) (2005)
  • Author(s): Suzuki T, Okada Y, Semba S, Orba Y, Yamanouchi S, Endo S, Tanaka S, Fujita T, Kuroda S, Nagashima K, Sawa H*
  • Journal Title: J Biol Chem 280 Pages: 24948-24956
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Dissociation of heterochromatin protein 1 from lamin B receptor induced by human polyomavirus agnoprotein : role in nuclear egress of viral particles. (* corresponding author) (2005)
  • Author(s): Okada Y, Suzuki T, Sunden Y, Orba Y, Kose S, Imamoto N, Takahashi H, Tanaka S, Hall WW, Nagashima K, Sawa H*
  • Journal Title: EMBO Rep 6 Pages: 452-457
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Isolation of a monoclonal antibody recognizing a cell-surface molecule as a receptor for JC virus. (* corresponding author) (2005)
  • Author(s): Henmi C, Sawa H*, Iwata H, Orba Y, Tanaka S, Nagashima K
  • Journal Title: Biochem Biophys Res Commun 327 Pages: 242-251
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Protection against influenza virus infection by intranasal administration of HA vaccine with chitin microparticles as an adiuvant. (2005)
  • Author(s): Hasegawa H, Ichinohe T, Strong P, Watanabe I, Ito S, Tamura S-i, Takahashi H, Sawa H, Chiba J, Kurata T, Sata T
  • Journal Title: JMed Virol 75 Pages: 130-136
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Variable patterns of varicella-zoster virus reactivation in Ramsay Hunt syndrome. (2004)
  • Author(s): Aizawa H, Ohtani F, Furuta Y, Sawa H, Fukuda S
  • Journal Title: J Med Virol 74 Pages: 355-360
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Investigation of simian virus 40 large T antigen in 18 autopsied malignant mesothelioma patients in Japan. (2004)
  • Author(s): Jin M, Sawa H^*, Suzuki T, Shimizu K, Makino Y, Tanaka S, Nojima T, Fujioka Y, Asamoto M, Suko N, Nagashima K
  • Journal Title: J Med Virol 74 Pages: 686-676
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Nuclear entry mechanism of the human polyomavirus JC virus like particle : role of importins and the nuclear pore complex. (2004)
  • Author(s): Qu Q, Sawa H^*, Suzuki T, Semba S, Henmi C, Okada Y, Tsuda M, Tanaka S, Atwood WJ, Nagashima K
  • Journal Title: J Biol Chem 279 Pages: 27735-27742
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Inhibition of virus production in JC virus-infected cells by postinfection RNA interference. (2004)
  • Author(s): Orba Y, Sawa H^*, Iwata H, Tanaka S, Nagashima K
  • Journal Title: J Virol 78 Pages: 7270-7273
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Nucleolin and the packaging signal, Ψ promote the budding of human immunodeficiency virus type-1 (HIV-1). (2004)
  • Author(s): Ueno T, Tokunaga K, Sawa H, Maeda M, Chiba J, Kojima A, Hasegawa H, Shoya Y, Sata T, Kutrata T, Takahashi H
  • Journal Title: Microbiol Immunol 48 Pages: 111-118
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Topoisomerase I dissociates human immunodeficiency virus type 1 reverse transcriptase from genomic RNAs. (2004)
  • Author(s): Takahashi H, Sawa H, Hasegawa H, Nagashima K, Sata T, Kurata T
  • Journal Title: Biochem Biophys Res Commun 313 Pages: 1073-1078
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Varicella-zoster virus DNA level and facial paralysis in Ramsay Hunt syndrome. (2004)
  • Author(s): Furuta Y, Aizawa H, Ohtani F, Sawa H, Fukuda S
  • Journal Title: Ann Otol Rhinol Laryngol 113 Pages: 700-705
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Investigation of simian virus 40 large T antigen in 18 autopsied malignant mesothelioma patients in Japan. (* corresponding author) (2004)
  • Author(s): Jin M, Sawa H*, Suzuki T, Shimizu K, Makino Y, Tanaka S, Nojima T, Fujioka Y, Asamoto M, Suko N, Nagashima K
  • Journal Title: J Med Virol 74 Pages: 668-676
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Nuclear entry mechanism of the human polyomavirus JC virus like particle : role of importins and the nuclear pore complex. (* corresponding author) (2004)
  • Author(s): Qu Q, Sawa H*, Suzuki T, Semba S, Henmi C, Okada Y, Tsuda M, Tanaka S, Atwood WJ, Nagashima K
  • Journal Title: J Biol Chem 279 Pages: 27735-27742
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Tbpoisomerase I dissociates human immunodeficiency virus type 1 reverse transcriptase from genomic RNAs. (2004)
  • Author(s): Takahashi H, Sawa H, Hasegawa H, Nagashima K, Sata T, Kurata T
  • Journal Title: Biochem Biophys Res Commun 313 Pages: 1073-1078
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Nucleolin and the packaging signal, ψ promote the budding of human immunodeficiency virus type-1 (HIV-1). (2004)
  • Author(s): Ueno T, Tokunaga K, Sawa H, Maeda M, Chiba J, Kojima A, Hasegawa H, Shoya Y, Sata T, Kutrata T, Takahashi H
  • Journal Title: Microbiol Immunol 48 Pages: 111-118
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Inhibition of virus production in JC virus-infected cells by postinfection RNA interference. (* corresponding author) (2004)
  • Author(s): Orba Y, Sawa H*, Iwata H, Tanaka S, Nagashima K
  • Journal Title: J Virol 78 Pages: 7270-7273
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Induction of Chromosomal Instability in Colonic Cells by the Human Polyomavirus JC Virus. (2003)
  • Author(s): Ricciardiello L, Baglioni M, Giovannini C, Pariali M, Cenacchi G, Ripalti A, Landini MP, Sawa H, Nagashima K, Frisque RJ, Goel A, Boland CR, Tognon M, Roda E, Bazzoli F
  • Journal Title: Cancer Res 63 Pages: 7256-7262
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Progressive multifocal leukoencephalopathy in a patient with X-linked agammaglobulinemia. (2003)
  • Author(s): Teramoto T, Kaneko H, Futano M, Sawa H, Nagashima K, Hirose Y, Kondo N
  • Journal Title: Scand J Infect Dis 35 Pages: 910-911
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Human topoisomerase I promotes HIV-1 proviral DNA synthesis : implications for the species specificity and cellular tropism of HIV-1 infection. (2003)
  • Author(s): Shoya Y, Tokunaga T, Sawa H, Maeda M, Ueno T, Yoshikawa T, Sata T, Kurata T, Hall WW, Cullen BR, Takahashi H
  • Journal Title: Proc Natl Acad Sci USA 100 Pages: 8442-8447
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Protection against influenza virus infection by intranasal administration of C3d-fused hemagglutinin. (2003)
  • Author(s): Watanabe I, Ross TM, Tamura Si, Ichinohe T, Ito S, Takahashi H, Sawa H, Chiba J, Kurata T, Sata T, Hasegawa H
  • Journal Title: Vaccine 21 Pages: 4532-4538
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Human topoisomerase I promotes HIV-1 proviral DNA synthesis : implications for the species specificity and cellular tropism of HIV-1 infection.q (2003)
  • Author(s): Shoya Y, Tokunaga T, Sawa H, Maeda M, Ueno T, Yoshikawa T, Sata T, Kurata T, Hall WW, Cullen BR, Takahashi H
  • Journal Title: Proc Natl Acad Sci USA 100 Pages: 8442-8447
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Induction of Chromosomal Instability in Colonic Cells by the Human Polvomavirus JC Virus. (2003)
  • Author(s): Ricciardiello L, Baglioni M, Giovannini C, Pariali M, Cenacchi G, Ripalti A, Landini MP, Sawa H, Nagashima K, Frisque RJ, Goel A, Boland CR, Tognon M, Roda E, Bazzoli F
  • Journal Title: Cancer Res 63 Pages: 7256-62
  • Description: 「研究成果報告書概要(欧文)」より
• [Book] Methods in Molecular Biology. DNA viruses, Methods and Protocols. (2005)
  • Author(s): Sawa H, Komagome R
  • Total Pages: 175-186
  • Publisher: Humana Press
  • Description: 「研究成果報告書概要(和文)」より
• [Patent(Industrial Property Rights)] JCウイルスのVP-1に対するsiRNA、およびそれを含有してなる医薬組成物 (2005)
  • Inventor(s): 長嶋 和郎, 澤 洋文, 大場 靖子
  • Industrial Property Rights Holder: 科学技術振興機構
  • Industrial Property Number: 特願2006-513677
  • Filing Date: 2005-04-26
  • Description: 「研究成果報告書概要(和文)」より
• [Patent(Industrial Property Rights)] JCウイルスの粒子形成阻害剤 (2004)
  • Inventor(s): 長嶋 和郎, 澤 洋文, 仙葉 愼吾
  • Industrial Property Rights Holder: 科学技術振興機構
  • Industrial Property Number: 特願2004-165083
  • Filing Date: 2004-06-02
  • Description: 「研究成果報告書概要(和文)」より
• [Patent(Industrial Property Rights)] 成人T細胞白血病モデルマウス (2004)
  • Inventor(s): 長谷川秀樹, 澤洋文他2名
  • Industrial Property Rights Holder: 森山雅美
  • Industrial Property Number: 特願2004-204169
  • Filing Date: 2004-07-12
  • Description: 「研究成果報告書概要(和文)」より
• [Patent(Industrial Property Rights)] HIV-I複製抑制剤 (2004)
  • Inventor(s): 長嶋 和郎, 澤 洋文, 高橋 秀宗
  • Industrial Property Rights Holder: 科学技術振興機構, 国立感染症研究所
  • Industrial Property Number: 特願2004-61237
  • Filing Date: 2004-03-04
  • Description: 「研究成果報告書概要(和文)」より