Design and delivery of plasmid vector for spatiotemporal control of the expression of therapeutic protein and siRNA

• Project/Area Number: 21390009
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Physical pharmacy
• Research Institution: Kyoto University
• Principal Investigator: TAKAKURA Yoshinobu 京都大学, 大学院・薬学研究科, 教授 (30171432)
• Co-Investigator(Kenkyū-buntansha): NISHIKAWA Makiya 京都大学, 大学院・薬学研究科, 准教授 (40273437)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥18,330,000 (Direct Cost: ¥14,100,000、Indirect Cost: ¥4,230,000); Fiscal Year 2011: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000); Fiscal Year 2010: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000); Fiscal Year 2009: ¥6,890,000 (Direct Cost: ¥5,300,000、Indirect Cost: ¥1,590,000)
• Keywords: ドラッグデリバリー / 遺伝子治療 / RNA干渉 / プラスミドDNA / 融合タンパク質 / デリバリー

Research Abstract

Interferon-γ(IFN-γ) was selected as a therapeutic protein and several IFN-γgene therapy systems were developed. Selection of proper promoters of IFN-γ-expressing vectors resulted in the regulation of the time profile of IFN-γtransgene expression. A fusion of IFN-γwith a protein that regulates the tissue distribution resulted in the spatiotemporal control of IFN-γafter in vivo gene transfer. This control led to an increase in the therapeutic effect of IFN-γ-based gene therapy on metastatic tumor with reduction in adverse effects.

Research Products

• [Journal Article] Constant and steady transgene expression of interferon-γby optimization of plasmid construct for safe and effective interferon-γgene therapy (2012)
  • Author(s): Ando M, Takahashi Y, Nishikawa M, Watanabe Y, Takakura Y.
  • Journal Title: J Gene Med Volume: 14 Issue: 4 Pages: 288-295
  • DOI: 10.1002/jgm.2616
  • Peer Reviewed
• [Journal Article] Prolonged circulation half-life of interferonγactivity by gene delivery of interferonγ-serum albumin fusion protein in mice (2011)
  • Author(s): Miyakawa N, Nishikawa M, Takahashi Y, Ando M, Misaka M, Watanabe Y, Takakura Y.
  • Journal Title: J Pharm Sci Volume: 100 Issue: 6 Pages: 2350-2357
  • DOI: 10.1002/jps.22473
  • Peer Reviewed
• [Journal Article] Increase in CpG DNA-induced inflammatory responses by DNA oxidation in macrophages and mice (2011)
  • Author(s): Yoshida H, Nishikawa M, Kiyota T, Toyota H, Takakura Y.
  • Journal Title: Free Radic Biol Med Volume: 51 Issue: 2 Pages: 424-431
  • DOI: 10.1016/j.freeradbiomed.2011.04.035
  • Peer Reviewed
• [Journal Article] 5'-Phosphate oligodeoxynucleotides enhance the phosphodiester-CpG DNA-induced inflammatory response in macrophages (2011)
  • Author(s): Yoshida H, Nishikawa M, Kiyota T, Uno S, Toyota H, Takahashi R, Narita M, Takakura Y.
  • Journal Title: Eur J Immunol Volume: 41 Issue: 2 Pages: 425-436
  • DOI: 10.1002/eji.201040396
  • Peer Reviewed
• [Journal Article] Saturation of transgene protein synthesis from mRNA in cells producing a large number of transgene mRNA (2011)
  • Author(s): Takahashi Y, Nishikawa M, Takiguchi N, Suehara T, Takakura Y.
  • Journal Title: Biotechnol Bioeng Volume: 108 Issue: 10 Pages: 2380-2389
  • DOI: 10.1002/bit.23179
  • NAID: 120005593565
  • Peer Reviewed
• [Journal Article] Inhibition of nuclear delivery of plasmid DNA and transcription by interferonγ: hurdles to be overcome for sustained gene therapy (2011)
  • Author(s): Zang L, Nishikawa M, Machida K, AndoM, Takahashi Y, Watanabe Y, Takakura Y.
  • Journal Title: Gene Ther Volume: 18 Issue: 9 Pages: 891-897
  • DOI: 10.1038/gt.2011.35
  • Peer Reviewed
• [Journal Article] 5'-Phosphate oligodeoxynucleotides enhance the phosphodiester-CpG DNA-induced inflammatory response in macrophages. (2011)
  • Author(s): Yoshida H., Nishikawa M., Takakura Y., et al.
  • Journal Title: European Journal of Immunology Volume: 41 Pages: 425-436
  • Peer Reviewed
• [Journal Article] Injection site-dependent induction of immune response by DNA vaccine : comparison of skin and spleen as a target for vaccination (2010)
  • Author(s): Guan X, Nishikawa M, Takemoto S, Ohno Y, Yata T, Takakura Y.
  • Journal Title: J Gene Med Volume: 12 Issue: 3 Pages: 301-309
  • DOI: 10.1002/jgm.1432
  • Peer Reviewed
• [Journal Article] Development of a novel Hsp70-based DNA vaccine as a multifunctional antigen delivery system (2010)
  • Author(s): Yamaoka A, Guan X, Takemoto S, Nishikawa M, Takakura Y.
  • Journal Title: J Control Release Volume: 142 Issue: 3 Pages: 411-415
  • DOI: 10.1016/j.jconrel.2009.11.005
  • Peer Reviewed
• [Journal Article] Sustained exogenous expression of therapeutic levels of IFN-gamma ameliorates atopic dermatitis in NC/Nga mice via Th1 polarization (2010)
  • Author(s): Hattori K, Nishikawa M, Watcharanurak K, Ikoma A, Kabashima K, Toyota H, Takahashi Y, Takahashi R, Watanabe Y, Takakura Y.
  • Journal Title: J Immunol Volume: 184 Issue: 5 Pages: 2729-2735
  • DOI: 10.4049/jimmunol.0900215
  • Peer Reviewed
• [Journal Article] Persistent interferon transgene expression by RNA interference-mediated silencing of interferon receptors (2010)
  • Author(s): Takahashi Y, Vikman E, Nishikawa M, Ando M, Watanabe Y, Takakura Y.
  • Journal Title: J Gene Med Volume: 12 Issue: 9 Pages: 739-746
  • DOI: 10.1002/jgm.1493
  • Peer Reviewed
• [Journal Article] Induction of Tumor-specific Immune Response by Gene Transfer of Hsp70-cell-penetrating Peptide Fusion Protein to Tumors in Mice (2010)
  • Author(s): Nishikawa M, Otsuki T, Ota A, Guan X, Takemoto S, Takahashi Y, Takakura Y.
  • Journal Title: Mol Ther Volume: 18 Issue: 2 Pages: 421-428
  • DOI: 10.1038/mt.2009.203
  • Peer Reviewed
• [Journal Article] Sustained exogenous expression of therapeutic levels of IFN-γ ameliorates atopic dermatitis in NC/Nga mice via Th1 polarization. (2010)
  • Author(s): Hattori K., Nishikawa M., Takahashi Y., Takakura Y., et al.
  • Journal Title: Journal of Immunology Volume: 184 Pages: 2729-2735
  • Peer Reviewed
• [Journal Article] Persistent interferon transgene expression by RNA interference-mediated silencing of interferon receptors. (2010)
  • Author(s): Takahashi Y.Nishikawa M., Takakura Y., et al.
  • Journal Title: Journal of Gene Medicine Volume: 12 Pages: 739-746
  • Peer Reviewed
• [Journal Article] Injection site-dependent induction of immune response by DNA vaccine : comparison of skin and spleen as a target for vaccination. (2010)
  • Author(s): Guan X., Nishikawa M., Takakura Y., et al.
  • Journal Title: Journal of Gene Medicine Volume: 12 Pages: 301-309
  • Peer Reviewed
• [Journal Article] Development of a novel Hsp70-based DNA vaccine as a multifunctional antigen delivery system (2010)
  • Author(s): Yamaoka A, et al.
  • Journal Title: Journal of Controlled Release 142 Pages: 411-415
  • Peer Reviewed
• [Journal Article] Injection site-dependent induction of immune response by DNA vaccine : comparison of skin and spleen as a target for vaccination (2010)
  • Author(s): Guan X, et al.
  • Journal Title: Journal of Gene Medicine 12 Pages: 301-309
  • Peer Reviewed
• [Journal Article] Induction of tumor-specific immune response by gene transfer of Hsp70-cell-penetrating peptide fusion protein to tumors in mice (2010)
  • Author(s): Nishikawa M, et al.
  • Journal Title: Molecular Therapy 18 Pages: 421-428
  • Peer Reviewed
• [Journal Article] Nonviral vector-mediated RNA interference : its gene silencing characteristics and important factors to achieve RNAi-based gene therapy Adv Drug Deliv Rev (2009)
  • Author(s): Takahashi Y, Nishikawa M, Takakura Y.
  • Journal Title: Advanced Drug Delivery Reviews Volume: 61 Issue: 9 Pages: 760-766
  • DOI: 10.1016/j.addr.2009.04.006
  • ISSN: 0169-409X
  • Peer Reviewed
• [Journal Article] Enhanced generation of cytotoxic T lymphocytes by increased cytosolic delivery of MHC class I epitope fused to mouse heat shock protein 70 via polyhistidine conjugation (2009)
  • Author(s): Takemoto S, Nishikawa M, Otsuki T, Yamaoka A, Maeda K, Ota A, Takakura Y.
  • Journal Title: J Control Release Volume: 135 Issue: 1 Pages: 11-18
  • DOI: 10.1016/j.jconrel.2008.11.024
  • Peer Reviewed
• [Journal Article] Quantitative and temporal analysis of gene silencing in tumor cells induced by small interfering RNA or short hairpin RNA expressed from plasmid vectors (2009)
  • Author(s): Takahashi Y, Yamaoka K, Nishikawa M, Takakura Y.
  • Journal Title: J Pharm Sci Volume: 98 Issue: 1 Pages: 74-80
  • DOI: 10.1002/jps.21398
  • Peer Reviewed
• [Journal Article] Enhanced CTL response by controlled intracellular trafficking of antigen in dendritic cells following DNA vaccination (2009)
  • Author(s): Isaji K, Kawase A, Matono M, Guan X, Nishikawa M, Takakura Y.
  • Journal Title: J Control Release Volume: 135 Issue: 3 Pages: 227-233
  • DOI: 10.1016/j.jconrel.2009.01.026
  • Peer Reviewed
• [Journal Article] Nonviral vector-mediated RNA interference : its gene silencing characteristics and important factors to achieve RNAi-based gene therapy (2009)
  • Author(s): Takahashi Y, et al.
  • Journal Title: Advanced Drug Delivery Reviews 61 Pages: 760-766
  • Peer Reviewed
• [Journal Article] Enhanced CTL response by controlled intracellular trafficking of antigen in dendritic cells following DNA vaccination (2009)
  • Author(s): Isaji K, et al.
  • Journal Title: Journal of Controlled Release 135 Pages: 227-233
  • Peer Reviewed
• [Journal Article] Enhanced CTL response by controlled intracellular trafficking of antigen in dendritic cells following DNA vaccination (2009)
  • Author(s): Takemoto S, et al.
  • Journal Title: Journal of Controlled Release 135 Pages: 11-18
  • Peer Reviewed
• [Journal Article] Quantitative and temporal analysis of gene silencing in tumor cells induced by small interfering RNA or short hairpin RNA expressed from plasmid vectors (2009)
  • Author(s): Takahashi Y, et al.
  • Journal Title: Journal of Pharmaceutical Sciences 98 Pages: 74-80
  • Peer Reviewed
• [Presentation] 核酸を基盤としたナノメディシンのデザインとデリバリー (2011)
  • Author(s): 高倉喜信
  • Organizer: 第20回DDSカンファランス
  • Place of Presentation: 静岡県コンベンションアーツセンター(静岡県)
  • Year and Date: 2011-09-16
• [Presentation] 核酸を基盤としたナノメディシンのデザインとデリバリー (2011)
  • Author(s): 高倉喜信
  • Organizer: 第20回DDSカンファランス
  • Place of Presentation: 静岡県コンベンションアーツセンター(静岡県)(招待講演)
  • Year and Date: 2011-09-16
• [Presentation] Kinetic comparison of gene silencing profile of siRNA and shRNA-expressing plasmid DNA in vivo (2011)
  • Author(s): Takahashi Y, Matsui Y, Nishikawa M, Takakura Y.
  • Organizer: 7th Annual Meeting of the Oligonucleotide Therapeutics Society
  • Place of Presentation: The Royal Library(Copenhagen, Denmark)
  • Year and Date: 2011-09-09
• [Presentation] Kinetic comparison of gene silencing profile of siRNA and shRNA-expressing plasmid DNA in vivo (2011)
  • Author(s): Takahashi Y, Matsui Y, Nishikawa M, Takakura Y
  • Organizer: 7th Annual Meeting of the Oligonucleotide Therapeutics Society
  • Place of Presentation: The Royal Library (Copenhagen, Denmark)
  • Year and Date: 2011-09-09
• [Presentation] 抗原タンパク質の発現プロファイル依存的な抗原特異的免疫応答の解析 (2011)
  • Author(s): 戎浦規文, 高橋有己, 西川元也, 高倉喜信
  • Organizer: 第21アンチセンスシンポジウム+第11回遺伝子・デリバリー研究会シンポジウム合同シンポジウム
  • Place of Presentation: 大阪大学コンベンションセンター(大阪府)
  • Year and Date: 2011-09-02
• [Presentation] Design and gene delivery of mouse interferonγgenetically fused with mouse serum albumin (2010)
  • Author(s): Miyakawa N, Nishikawa M, Takahashi Y, Watanabe Y, Takakura Y.
  • Organizer: Pharmaceutical Sciences World Congress
  • Place of Presentation: Morial convention center(New Orleans, USA)
  • Year and Date: 2010-11-16
• [Presentation] Interferonγ-mediated inhibition of nuclear entry of plasmid vector and mRNA transcription from the vector : an obstacle for repeated injections of interferonγ-expressing plasmid vector (2010)
  • Author(s): Zang L, Nishikawa M, Machida K, Ando M, Takahashi Y, Takakura Y.
  • Organizer: Pharmaceutical Sciences World Congress
  • Place of Presentation: Morial convention center(New Orleans, USA)
  • Year and Date: 2010-11-16
• [Presentation] Design and gene delivery of mouse interferon γ genetically fused with mouse serum albumin. (2010)
  • Author(s): Miyakawa N., et al.
  • Organizer: 2010 Pharmaceutical Sciences World Congress
  • Place of Presentation: Morial convention center(New Orleans, USA)
• [Presentation] Interferon γ-mediated inhibition of nuclear entry of plasmid vector and mRNA transcription from the vector:an obstacle for repeated injections of interferon γ-expressing plasmid vector. (2010)
  • Author(s): Zang L., et al.
  • Organizer: 2010 Pharmaceutical Sciences World Congress
  • Place of Presentation: Morial convention center(New Orleans, USA)
• [Presentation] Contradictory effects of gene silencing of b-catenin in melanoma cells on proliferation and metastasis (2009)
  • Author(s): 高橋有己, 西川元也, 高倉喜信
  • Organizer: 第68回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜(神奈川県)
  • Year and Date: 2009-10-03
• [Presentation] Contradictory effects of gene silencing of β-catenin in melanoma cells on proliferation and metastasis (β-catenin 遺伝子発現抑制によるがん細胞増殖抑制効果と転移促進) (2009)
  • Author(s): Takahashi Y, et al.
  • Organizer: 第68回日本癌学会学術総会
  • Place of Presentation: 横浜