Construction of human artificial chromosome for gene therapy of Duchenne muscular dystrophy

• Project/Area Number: 18390107
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Human genetics
• Research Institution: Tottori University
• Principal Investigator: OSHIMURA Mitsuo Tottori University, 大学院・医学系研究科, 教授 (20111619)
• Co-Investigator(Kenkyū-buntansha): KATO Motonobu 鳥取大学, 医学部, 助教 (00273904) ; KAZUKI Yasuhiro 鳥取大学, 大学院・医学系研究科, 助教 (90403401)
• Project Period (FY): 2006 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥13,160,000 (Direct Cost: ¥11,000,000、Indirect Cost: ¥2,160,000); Fiscal Year 2008: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2007: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2006: ¥3,800,000 (Direct Cost: ¥3,800,000)
• Keywords: 遺伝子治療 / 染色体工学 / ヒト人工染色体 / 筋ジストロフィー / 間葉系幹細胞 / ES細胞

Research Abstract

デュシャンヌ型筋ジストロフィー(DMD)の遺伝子治療を目指して、2. 4Mb におよぶジストロフィン(DMD)遺伝子ゲノム領域を染色体改変技術を利用した染色体転座型クローニング法でHAC ベクター上にクローニングすることに成功した(DYS-HAC ベクターとよぶ)。このDYS-HACベクターを保持するキメラマウスにおいて、DMD 遺伝子の発現は組織特異的かつ筋肉の細胞膜周辺に発現することが示された。さらにヒト細胞でのDYS-HAC ベクターは非常に安に維持されることが示された。以上のことより、本研究で作製したDYS-HAC ベクターはDMD の遺伝子治療に利用できることが示された。

Research Products

• [Journal Article] A highly stable and non-integrated human artificial chromosome(HAC)containing the 2.4Mb entire human dystrophin gene (2009)
  • Author(s): Hoshiya H.
  • Journal Title: Mol. Therapy. 17 Pages: 309-317
  • Peer Reviewed
• [Journal Article] Enhanced apoptosis during early neiironal differentiation in mouse ES cells with autosomal imbalance. (2009)
  • Author(s): Kai Y.
  • Journal Title: Cell Res. 19 Pages: 247-258
  • Peer Reviewed
• [Journal Article] Transfer human artificial chromosome vectors into stem cells (2008)
  • Author(s): Oshimura M., Katoh M.
  • Journal Title: Reproductive BioMedicine Online Vol.16 No.1 Pages: 57-69
• [Journal Article] Correction of a genetic defect in multipotentGermline stem cells using a human artificial chromosome (2008)
  • Author(s): Kazuki Y., Hoshiya H., Kai Y., Abe S., Takiguchi M., Osaki M.,Kawazoe S., Katoh M., Kanatsu-Shinohara M., Inoue K., Kajitani N., Yoshino N., Shirayoshi Y., Ogura A., Shinohara T., BarrettJC., Oshimura M.
  • Journal Title: Gene Therapy 15 Pages: 617-24
• [Journal Article] A highly stable and non-integrated human artificial chromosome(HAC) containing the 2.4Mb entire human dystrophin gene (2008)
  • Author(s): Hoshiya H., Kazuki Y., Abe S., Takiguchi M., Kajitani N., Watanabe Y., Yoshino T.,Shirayoshi Y., Higaki K., Messina G., Cossu G., Oshimura M.,
  • Journal Title: Mol. Therapy
• [Journal Article] Telomerase-mediated life-span extension of human primary fibroblasts by human artificial chromosome (HAC) vector (2008)
  • Author(s): Shitara S, Kakeda M, Nagata K, Hiratsuka M, Sano A, Osawa K, Okazaki A, Katoh M, Kazuki Y, Oshimura M, Tomizuka K
  • Journal Title: Biochem Biophys Res Commun Volume 369, number 3 Pages: 807-11
• [Journal Article] Telomerase-mediated life-span extension of human primary fibroblasts by human artificial chromosome(HAC)vector. (2008)
  • Author(s): Shitara S
  • Journal Title: Biochem Biophys Res Commun. 369 Pages: 807-811
  • Peer Reviewed
• [Journal Article] Correction of a genetic defect in multipotent germline stem cells using a human artificial chromosome. (2008)
  • Author(s): Kazuki Y
  • Journal Title: Gene Ther. 15 Pages: 617-624
  • Peer Reviewed
• [Journal Article] Transfer of human artificial chromosome vectors into stem cells. (2008)
  • Author(s): Oshimura M
  • Journal Title: Reprod Biomed Online. 16 Pages: 57-69
  • Peer Reviewed
• [Journal Article] Human artificial chromosome vectors meet stem cells : new prospects for gene delivery (2006)
  • Author(s): Ren X, Tahimic CG, Katoh M, Kurimasa A, Inoue T, Oshimura M
  • Journal Title: Stem Cell Rev 2 Pages: 43-50
• [Journal Article] Human artificial choromosome vectors meet stem cells : new prospects for gene delivery. (2006)
  • Author(s): Ren X
  • Journal Title: Stem Cell 2・1 Pages: 43-50
• [Presentation] 染色体工学技術を用いたヒト型モデル動物の作製と応用 (2009)
  • Author(s): 香月康宏
  • Organizer: 第129回日本薬学会年会
  • Place of Presentation: 京都
  • Year and Date: 2009-03-26
• [Presentation] A novel human artificial chromosome (HAC)for gene therapy and animal transgenesis (2008)
  • Author(s): Kazuki Y
  • Organizer: ASGT 11^<th> annual meeting
  • Place of Presentation: USA, Boston
  • Year and Date: 2008-05-30
• [Presentation] A human artificial chromosome (HAC)vector with about 2.4 Mb-human Dystrophin genome including native expression control elements (2008)
  • Author(s): Hoshiya H
  • Organizer: ASGT 11^<th> annual meeting
  • Place of Presentation: USA, Boston
  • Year and Date: 2008-05-30
• [Presentation] 新規ヒト人工染色体ベクターの構築 : サブテロメアを用いた高発現ベクター系 (2008)
  • Author(s): 掛田実、永田恵子、佐野暁子、大澤加奈子、岡崎晃代、設楽真吾、香月康宏、押村光雄、富塚一磨
  • Organizer: 第6回幹細胞シンポジウム
  • Place of Presentation: 東京
• [Presentation] A human artificial chromosome (HAC) vector with about 2.4 Mb-human Dystrophin genome including native expression control elements (2008)
  • Author(s): Hoshiya H., Kazuki Y., Abe S., Kai Y., Takiguchi M., Iida Y., Watanabe Y., Osaki M., Kajitani N., Yoshino T., Shirayoshi Y. and Oshimura M.
  • Organizer: ASGT 11^<th> annual meeting
  • Place of Presentation: U.S.A.
• [Presentation] A novel human artificial chromosome (HAC) for gene therapy and animal transgenesis (2008)
  • Author(s): Kazuki Y., Hoshiya H., Abe S., Takiguchi M., Iida Y., Watanabe Y., Osaki M., Katoh M., Kajitani N., Yoshino T., Shirayoshi Y., Hiratsuka M., Kurimasa K., and Oshimura M.
  • Organizer: ASGT 11th annual meeting
  • Place of Presentation: U.S.A.
• [Presentation] 遺伝子治療に向けた新規遺伝子治療用ヒト人工染色体ベクターの開発 (2008)
  • Author(s): 香月康宏, 星谷英寿, 甲斐義輝, 阿部智志, 滝口正人, 飯田雄一, 渡辺義徳, 尾崎克彦, 梶谷尚世, 吉野とう子, 白吉安昭, 富松望, 平塚正治, 栗政明弘, 押村光雄
  • Organizer: 第7回日本再生医療学会総会
  • Place of Presentation: 名古屋
• [Presentation] 多能性精原幹細胞(mGS)とヒト人工染色体ベクターを用いた欠損型遺伝子治療を目指して (2008)
  • Author(s): 阿部智志、香月康宏、星谷英寿、甲斐義輝、滝口正人、尾崎克彦、川添真史郎、加藤基伸、篠原美都、井上貴美子、梶谷尚世、吉野とう子、白吉安昭、小倉淳郎、篠原隆司、押村光雄
  • Organizer: 第7回日本再生医療学会総会
  • Place of Presentation: 名古屋
• [Presentation] ジストロフィンゲノムDNA(2.5Mb)を搭載したHAC ベクター : 遺伝子治療を目指して (2008)
  • Author(s): 星谷英寿、香月康宏、甲斐義輝、阿部智志、滝口正人、飯田雄一 ,渡辺義徳, 尾崎克彦、梶谷尚世,吉野とう子,白吉安昭 ,押村光雄
  • Organizer: 第7回日本再生医療学会総会
  • Place of Presentation: 名古屋
• [Presentation] Genetic restoration of ataxia telangiectasia cells using a human artificial chromosome mediated-genomic cloning system (2007)
  • Author(s): Kazuki Y
  • Organizer: ASGT 10th annual meeting
  • Place of Presentation: USA
  • Year and Date: 2007-06-01
• [Presentation] Functional restoration of a genetic defect in multipotent germline stem cells by human artificial chromosome containing a genomic insert (2006)
  • Author(s): Kazuki Y., Hoshiya, H., Kai Y., Abe S., Takiguchi M., Osaki M., Kanatsu-Shinohara M., Kajitani N., ShirayoshiY., Hratsuka M., Ogura A., Shinohara T.and Oshimura, M
  • Organizer: ASGT 9^<th> annual meeting
  • Place of Presentation: U.S.A.
• [Book] 幹細胞とヒト人工染色体ベクターの出会いVol.19, No.1 (2007)
  • Author(s): 香月康宏、押村光雄
  • Publisher: アニテックス
• [Book] ヒト人工染色体ベクター導入と遺伝子・再生医療への可能性vol.10No.4 (2006)
  • Author(s): 香月康宏、押村光雄
  • Publisher: 分子呼吸器病
• [Book] 11Vol.5 No.4 (2006)
  • Author(s): 平塚正治、押村光雄
  • Publisher: 再生医療
• [Patent(Industrial Property Rights)] ヒト人工染色体(HAC)ベクター及びヒト人工染色体(HAC)を有するヒト細胞医薬 (2007)
  • Inventor(s): 掛田実、富塚一磨、押村光雄、香月康宏
  • Filing Date: 2007-07-06
  • Overseas
• [Patent(Industrial Property Rights)] ヒト人工染色体(HAC)ベクター及びヒト人工染色体(HAC)を有するヒト細胞医薬 (2007)
  • Inventor(s): 掛田実、富塚一磨、押村光雄、香月康宏
  • Industrial Property Rights Holder: キリンファーマ株式会社
  • Filing Date: 2007-07-06
  • Overseas
• [Patent(Industrial Property Rights)] 内在遺伝子を含まないヒト人工染色体ベクター (2006)
  • Inventor(s): 押村光雄、香月康宏、松岡隆之
  • Industrial Property Number: 2006-127372
  • Filing Date: 2006-05-02
• [Patent(Industrial Property Rights)] 内在遺伝子を含まないヒト人工染色体ベクター (2006)
  • Inventor(s): 押村光雄, 香月康宏, 松岡隆之
  • Industrial Property Rights Holder: 鳥取大学, 有限株式会社chromocenter
  • Industrial Property Number: 2006-127372
  • Filing Date: 2006-05-02