Toward a development of the novel drug-screening system based on monitoring autophagy dynamics

• Project/Area Number: 24650189
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Nerve anatomy/Neuropathology
• Research Institution: Tokai University
• Principal Investigator: HADANO Shinji 東海大学, 医学部, 教授 (60281375)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2014: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2013: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2012: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 脳神経疾患 / 脳・神経 / トランスレーショナルリサーチ / 動物 / 神経科学 / 脳･神経

Outline of Final Research Achievements

The autophagy-endolysosomal system is an evolutionally conserved degradation system that is tightly linked to a wide variety of physiological processes. Dysfunction of this system is associated with many neurodegenerative diseases including amyotrophic lateral sclerosis (ALS). In this study, we sought to generate a novel analytical tool for the autophagy-endolysosomal system in vivo. First, we generated the expression construct encoding mKeimaRed_LC3B chimeric protein, and successfully detected the bimodal excitation spectrum of mKeimaRed_LC3B in HeLa cells under starved conditions. Then, we microinjected it into fertilized eggs from C57BL/6 mice, and transplanted them into the oviduct of pseudopregnant recipient mice. As a result, we have successfully established 2 lines of transgenic mouse expressing mKeima-Red_LC3B, particularly in heart, skeletal muscle and the central nervous system. These mice could become useful tools for development of the new drug-screening systems.

Research Products

• [Journal Article] Six SQSTM1 mutations in a Chinese amyotrophic lateral sclerosis cohort. (2015)
  • Author(s): Yang, Y., Tang, L., Zhang, N., Pan, L., Hadano, S., and Fan, D. -S.
  • Journal Title: Amyotroph. Lateral Scler. Frontotempo. Degen. Volume: in press Issue: 5-6 Pages: 378-384
  • DOI: 10.3109/21678421.2015.1009466
  • Peer Reviewed
• [Journal Article] SQSTM1 mutations in Han Chinese populations with sporadic amyotrophic lateral sclerosis. (2014)
  • Author(s): Chen, Y. P., Zheng, Z. Z., Chen, X. P., Huang, R., Yang, Y., Yuan, L. X., Pan, L., Hadano, S., and Shang, H. -F.
  • Journal Title: Neurobiol. Aging Volume: 35 Issue: 3 Pages: 726.e7-726.e9
  • DOI: 10.1016/j.neurobiolaging.2013.09.008
  • Peer Reviewed
• [Journal Article] Different Human Copper-Zinc Superoxide Dismutase Mutants, SOD1G93A and SOD1H46R, Exert Distinct Harmful Effects on Gross Phenotype in Mice (2012)
  • Author(s): Lei Pan
  • Journal Title: PLoS One Volume: 7巻 Issue: 3 Pages: e33409-e33409
  • DOI: 10.1371/journal.pone.0033409
  • Peer Reviewed
• [Journal Article] A novel small-molecule, N-(4-(2-pyridyl)(1,3-thiazol-2-yl))-2-(2,4,6-trimethylphenoxy)acetamide, selectively protects against oxidative stress-induced cell death by activating the Nrf2-ARE pathway: Therapeutic implications for ALS. (2012)
  • Author(s): Kanno, T.
  • Journal Title: Free Radical Biol. Med. Volume: 53 Issue: 11 Pages: 2028-2042
  • DOI: 10.1016/j.freeradbiomed.2012.09.010
  • Peer Reviewed
• [Journal Article] Dysregulation of the autophagy-endolysosomal system in amyotrophic lateral sclerosis and related motor neuron diseases (2012)
  • Author(s): Otomo A, Pan L, and Hadano S
  • Journal Title: Neurol. Res. Int Volume: 2012 Pages: 498428-498428
  • DOI: 10.1155/2012/498428
  • Peer Reviewed
• [Presentation] p62/SQSTM1 deficiency accelerates motor neuron degeneration in SOD1H46R transgenic mice. (2014)
  • Author(s): Pan, L., Otomo, A., Koike, M., Uchiyama, Y., Aoki, M., Abe, K., Ishii, T., Yanagawa, T., Shang, H. -F., Yoshii, F., and Hadano, S.
  • Organizer: 25th International Symposium on ALS/MND
  • Place of Presentation: Brussels, Belgium
  • Year and Date: 2014-12-05 – 2014-12-07
• [Presentation] オートファジー動態の可視化を目的とした新規トランスジェニックマウスの作出 (2014)
  • Author(s): 秦野伸二、王婷、田中政之、林英樹、荻原早苗、岡田千沙、伊藤誠敏、福西菜穂子、飯田裕美、中村彩花、大友麻子、大塚正人
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] 筋萎縮性側索硬化症原因遺伝子産物ALS2に結合する新規調節因子Rab30の機能解析 (2014)
  • Author(s): 小野寺和歌奈、大友麻子、福田光則、秦野伸二
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] ALS2及び新規ALS2結合低分子量Gタンパク質Rab17の細胞内局在解析 (2014)
  • Author(s): 小野鈴花、大友麻子、福田光則、秦野伸二
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] Loss of p62/SQSTM1, an autophagy substrate, aggravates motor dysfunction in a SOD1H46R-expressing mouse ALS model. (2014)
  • Author(s): Hadano, S., Pan, L., Otomo, A., Abe, K., Koike, M., Uchiyama, Y., Aoki, M., Ishii, T., Yanagawa, T., Shang, H. -F., and Yoshii, F.
  • Organizer: 第37回日本神経科学大会
  • Place of Presentation: 横浜
  • Year and Date: 2014-09-11 – 2014-09-13
• [Presentation] オートファジー関連因子p62/SQSTM1の機能喪失はALSマウスモデルの疾患症状を悪化させる (2014)
  • Author(s): 秦野伸二、潘雷、大友麻子、阿部幸一郎、小池正人、内山安男、青木正志、吉井文均、石井哲郎、柳川徹
  • Organizer: 第61回日本実験動物学会総会
  • Place of Presentation: 札幌
  • Year and Date: 2014-05-15 – 2014-05-17
• [Presentation] Loss of p62/SQSTM1 exacerbates motor dysfunction in a mutant SOD1-expressing mouse ALS model. (2013)
  • Author(s): Pan, L., Otomo, A., Abe, K., Ogawa, H., Chiba, T., Koike, M., Uchiyama, Y., Aoki, M., Yoshii, F., Ishii, T., Yanagawa, T., and Hadano, S.
  • Organizer: 24th International Symposium on ALS/MND
  • Place of Presentation: Milan, Italy
• [Presentation] SQSTM1 mutations in sporadic Chinese patients with amyotrophic lateral sclerosis. (2013)
  • Author(s): Chen, Y., Chen, X., Huang, R., Zheng, Z., Wei, Q., Guo, X., Pan, L., Hadano, S., and Shang, H.
  • Organizer: 24th International Symposium on ALS/MND
  • Place of Presentation: Milan, Italy
• [Presentation] ALSモデルマウスにおけるIP6K2の病態への関連性について（第２報） (2013)
  • Author(s): 森谷祐介、永田栄一郎、潘雷、佐藤忠之、小川温子、秦野伸二、瀧澤俊也
  • Organizer: 第54回日本神経学会学術大会
  • Place of Presentation: 東京
• [Presentation] 筋萎縮性側索硬化症におけるイノシトール６リン酸キナーゼ２の役割 (2013)
  • Author(s): 永田栄一郎、森谷祐介、藤井奈津子、小原さおり、佐藤忠之、高尾昌樹、潘雷、小川温子、秦野伸二、三原盤、瀧澤俊也
  • Organizer: Neuro2013：第36回日本神経科学会大会/第56回日本神経化学会大会/第23回日本神経回路学会大会
  • Place of Presentation: 京都
• [Presentation] 低分子量Gタンパク質Rab5の活性化因子、ALS2とRabGEF1は、EEA1-LC3陽性小胞の成熟と分解効率を調節する (2013)
  • Author(s): 大友麻子、潘雷、小川温子、平塚結衣、秦野伸二
  • Organizer: Neuro2013：第36回日本神経科学会大会/第56回日本神経化学会大会/第23回日本神経回路学会大会
  • Place of Presentation: 京都
• [Presentation] SQSTM1遺伝子変異が筋萎縮性側索硬化症（ALS）の発症に及ぼす影響 (2013)
  • Author(s): 秦野伸二
  • Organizer: 第7回オートファジー研究会
  • Place of Presentation: 掛川
• [Presentation] マウスモデルを用いたALS疾患症状の調節要因に関する研究. (2013)
  • Author(s): 秦野伸二
  • Organizer: 東海大学総合医学研究所第４回シンポジウム
  • Place of Presentation: 霞ヶ関/東京
• [Presentation] Dysregulation of the autophagy-endolysosomal system in motor neuron diseases. (2012)
  • Author(s): Hadano, S.
  • Organizer: BIT’s 3rd Annual World Congress on NeuroTalk-2012
  • Place of Presentation: Beijing, China
• [Presentation] Dysregulation of the autophagy-endolysosomal system in ALS and FTD. (2012)
  • Author(s): Hadano, S.
  • Organizer: The 15th National Conference of Neurology
  • Place of Presentation: Guangzhou, China
  • Invited
• [Presentation] Different human copper-zinc superoxide dismutase mutants, SOD1G93A and SOD1H46R, exert distinct harmful effects on gross phenotype in mice. (2012)
  • Author(s): Pan, L.
  • Organizer: The 15th National Conference of Neurology
  • Place of Presentation: Guangzhou, China
• [Presentation] Role of inositol hexakisphosphate kinase 2 in a mouse model for amyotrophic lateral sclerosis. (2012)
  • Author(s): Moriya, Y.
  • Organizer: Neuroscience 2012
  • Place of Presentation: New Orleans, Louisiana, USA
• [Presentation] Different human copper-zinc superoxide dismutase mutants, SOD1G93A and SOD1H46R, exert distinct harmful effects on gross phenotype in mice. (2012)
  • Author(s): Pan, L.
  • Organizer: 23rd International Symposium on ALS/MND
  • Place of Presentation: Chicago, Illinois, U.S.A.
• [Presentation] Bromocriptine retards disease progression in an ALS mouse model via suppression of glial inflammation. (2012)
  • Author(s): Tanaka, K.
  • Organizer: 23rd International Symposium on ALS/MND
  • Place of Presentation: Chicago, Illinois, U.S.A.
• [Presentation] ALSモデルマウスにおけるイノシトール6リン酸キナーゼ2（IP6K2）の病態への関連性について. (2012)
  • Author(s): 森谷祐介
  • Organizer: 第53回日本神経学会学術大会
  • Place of Presentation: 東京
• [Presentation] 家族性ALS原因遺伝子産物ALS2の機能喪失がオートファジー系に及ぼす影響. (2012)
  • Author(s): 秦野伸二
  • Organizer: 第11回生体機能研究会
  • Place of Presentation: 箱根/神奈川
• [Presentation] 低分子量Gタンパク質Rab5の活性化因子、ALS2とRabGEF1は、EEA1-LC3要請小胞の形成と成熟を 調節する. (2012)
  • Author(s): 大友麻子
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡
• [Presentation] Molecular pathogenesis of ALS/MNDs: Dysfunction of autophagy-endolysosomal system.
  • Author(s): Hadano, S.
  • Organizer: Fujian Conference of Neurology on Neurodegenerative Diseases
  • Place of Presentation: Fuzhou, China
  • Invited
• [Presentation] 細胞内エンドソーム動態と神経変性
  • Author(s): 秦野伸二
  • Organizer: 厚生労働科学研究費補助金難治性疾患等克服研究事業神経変性疾患に関する調査研究班、「病態に根ざしたALSの新規治療法開発」分科班、平成25年度ワークショップ
  • Place of Presentation: 東京
  • Invited
• [Presentation] QSTM1遺伝子変異が筋萎縮性側索硬化症（ALS）の発症に及ぼす影響
  • Author(s): 秦野伸二
  • Organizer: 東海大学総合医学研究所第17回公開研究報告会
  • Place of Presentation: 伊勢原
• [Book] ALS2/Alsin、アクチュアル脳・神経疾患の臨床「すべてがわかるALS・運動ニューロン疾患」IV. ALSの病態関連遺伝子と遺伝子変異、辻省次、祖父江元編 (2013)
  • Author(s): 大友麻子、白川健太郎、宮嶋裕明、秦野伸二
  • Publisher: 中山書店