Molecular mechanism for the maintenance of proteostasis in motor neurons

• Project/Area Number: 26290018
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Nerve anatomy/Neuropathology
• Research Institution: Tokai University
• Principal Investigator: HADANO SHINJI 東海大学, 医学部, 教授 (60281375)
• Co-Investigator(Kenkyū-buntansha): 大友 麻子 東海大学, 医学部, 助教 (50535226)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥16,770,000 (Direct Cost: ¥12,900,000、Indirect Cost: ¥3,870,000); Fiscal Year 2016: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2015: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2014: ¥7,150,000 (Direct Cost: ¥5,500,000、Indirect Cost: ¥1,650,000)
• Keywords: 脳神経疾患 / 脳・神経 / 神経科学 / 遺伝子発現 / 蛋白質分解 / 酸化ストレス / 運動ニューロン / ALS / 遺伝子 / 蛋白質

Outline of Final Research Achievements

ALS is a fatal neurodegenerative disorder characterized by a selective loss of motor neurons. Multiple toxicity pathways, such as oxidative stress and misfolded protein accumulation, are implicated in the pathogenesis of ALS. We generated SOD1H46R mice either on a Nfe2l2-KO, Sqstm1-KO, or Sqstm1/Als2-double KO background. Loss of SQSTM1 but not NFE2L2 exacerbated disease symptoms. A simultaneous inactivation of SQSTM1 and ALS2 further accelerated the onset of disease. Absence of SQSTM1 accelerated motor neuron degeneration with accompanying the preferential accumulation of ubiquitin-positive aggregates in spinal neurons. Surprisingly, loss of SQSTM1 rather suppressed the accumulation of insoluble polyubiquitinated proteins, suggesting that the selective accumulation of aggregates in neurons might be more insulting than the insoluble proteins. Collectively, SQSTM1 and ALS2 have distinct but additive protective roles against mutant SOD1-mediated toxicity by modulating proteostasis.

Research Products

• [Int'l Joint Research] Sichuan University(China)
• [Int'l Joint Research] Sichuan University/Pekin University(China)
• [Journal Article] Inositol hexakisphosphate kinase 2 is a pre-symptomatic biomarker for amyotrophic lateral sclerosis. (2017)
  • Author(s): Moriya, Y., Nagata, E., Fujii, N., Satoh, T., Ogawa, H., Hadano, S., and Takizawa, S.
  • Journal Title: Tokai J. Exp. Clin. Med. Volume: 42 Pages: 13-18
  • Peer Reviewed
• [Journal Article] Mutation screening of the CHCHD10 gene in Chinese patients with amyotrophic lateral sclerosis. (2017)
  • Author(s): Zhou, Q. Q., Chen, Y. P., Wei, Q. -Q., Cao, B., Wu, Y., Zhao, B., Ou, R. W., Yang, J., Chen, X. P., Hadano, S., and Shang, H. -F.
  • Journal Title: Mol. Neurobiol Volume: 印刷中 Issue: 5 Pages: 3189-3194
  • DOI: 10.1007/s12035-016-9888-0
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Functional links between SQSTM1 and ALS2 in the pathogenesis of ALS: Cumulative impact on the protection against mutant SOD1-mediated motor dysfunction in mice (2016)
  • Author(s): Hadano, S., Mitsui, S., Pan, L., Otomo, A., Kubo, M., Sato, K., Ono, S., Onodera, W., Abe, K., Koike, M., Uchiyama, Y., Aoki, M., Warabi, E., Yamamoto, M., Ishii, T., Yanagawa, T., Shang, HF., Yoshii, F
  • Journal Title: Hum. Mol. Genet. Volume: 25 Issue: 15 Pages: 3321-3340
  • DOI: 10.1093/hmg/ddw180
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Aberration of miRNAs expression in leukocytes from sporadic amyotrophic lateral sclerosis. (2016)
  • Author(s): Chen, Y. P., Wei, Q. Q., Chen, X. P., Li, C. Y., Cao, B., Ou, R. W., Hadano, S., and Shang, H. -F.
  • Journal Title: Front. Mol. Neurosci. Volume: 9
  • DOI: 10.3389/fnmol.2016.00069
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition) (2016)
  • Author(s): Klionsky DJ., Abdelmohsen K., Abe A., Abedin MJ.,Inomata M., et al.
  • Journal Title: Autophagy Volume: 12(1) Issue: 1 Pages: 1-222
  • DOI: 10.1080/15548627.2015.1100356
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [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
• [Presentation] Dichotomous effect of systemic overexpression of SQSTM1 on disease onset and post-onset survival in SOD1H46R mice. (2017)
  • Author(s): Mitsui, S., Otomo, A., Nozaki, M., Ono, S., Sato, K., Shirakawa, R., Adachi, H., Aoki, M., Sobue, G., Shang, H. -F. and Hadano, S.
  • Organizer: 第40回日本神経科学大会
  • Place of Presentation: 幕張メッセ（千葉県・千葉市）
  • Year and Date: 2017-07-20
• [Presentation] The ALS2-interacting protein Rab30 is a possible regulator for vesicular trafficking between early endosome and the Golgi apparatus. (2017)
  • Author(s): Otomo, A., Matsui, K., Onodera, W., Sugiyama, J., Ishida, T., Sato, K., Mitsui, S., Shirakawa, R., Nozaki, M., Ono, S., Fukuda, M., and Hadano, S.
  • Organizer: 第40回日本神経科学大会
  • Place of Presentation: 幕張メッセ（千葉県・千葉市）
  • Year and Date: 2017-07-20
• [Presentation] 軸索輸送現象の定量評価に向けた開放型マイクロデバイスの開発および評価 (2017)
  • Author(s): 横山奨，串田隆志，石田智之，大友麻子，秦野伸二，木村啓志
  • Organizer: シンポジウム「細胞アッセイ技術の現状と将来」、細胞アッセイ研究会
  • Place of Presentation: 東京大学生産技術研究所コンベンションホール（東京都・目黒区）
• [Presentation] Alterations of oligomeric states and subcellular localization of ALS2 mutants underlie the pathogenesis of ALS2-linked motor neuron diseases. (2016)
  • Author(s): Sato, K., Suzuki-Utsunomiya, K., Hiratsuka, Y., Mitsui, S., Ono, S., Otomo, A., and Hadano, S.
  • Organizer: 27th International Symposium on ALS/MND
  • Place of Presentation: Dublin (Republic of Ireland)
  • Year and Date: 2016-12-07
  • Int'l Joint Research
• [Presentation] Systemic overexpression of SQSTM1 accelerates age of disease onset and reduces survival in SOD1H46R mice. (2016)
  • Author(s): Mitsui, S., Otomo, A., Nozaki, M., Ono, S., Sato, K., Shirakawa, R., Adachi, H., Aoki, M., Sobue, G., Shang, H. -F., and Hadano, S.
  • Organizer: 27th International Symposium on ALS/MND
  • Place of Presentation: Dublin (Republic of Ireland)
  • Year and Date: 2016-12-07
  • Int'l Joint Research
• [Presentation] Analysis of axonal transport in cultured neurons derived from an ALS mouse model by using the microfluidic cell culture system. (2016)
  • Author(s): Otomo, A., Araki, R., Ishida, T., Shirakawa, R., Mitsui, S., Ono, S., Sato, K., Yokoyama, S., Kimura, H., and Hadano, S.
  • Organizer: 27th International Symposium on ALS/MND
  • Place of Presentation: Dublin (Republic of Ireland)
  • Year and Date: 2016-12-07
  • Int'l Joint Research
• [Presentation] ALS患者由来SQSTM1/p62変異体の神経細胞における局在と機能の解析 (2016)
  • Author(s): 白川涼平、濱祐太郎、大友麻子、Shang, H. -F.、秦野伸二
  • Organizer: 第39回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜（神奈川県・横浜市）
  • Year and Date: 2016-11-30
• [Presentation] マイクロ流体デバイスを用いたALS疾患モデルの解析 (2016)
  • Author(s): 石田智之、荒木良介、大友麻子、横山奨、串田隆志、杉山純也、秦野伸二、木村啓志
  • Organizer: 第39回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜（神奈川県・横浜市）
  • Year and Date: 2016-11-30
• [Presentation] 新規ALS2結合候補分子Rab30の発現及び細胞内動態解析 (2016)
  • Author(s): 松井香奈、小野寺和歌奈、大友麻子、福田光則、秦野伸二
  • Organizer: 第39回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜（神奈川県・横浜市）
  • Year and Date: 2016-11-30
• [Presentation] SQSTM1の全身性高発現は変異SOD1発現ALSマウスモデルの発症を早める (2016)
  • Author(s): 三井駿、大友麻子、野崎昌久、小野鈴花、佐藤海、白川涼平、足立弘明、青木正志、祖父江元、Shang, H. -F.、秦野伸二
  • Organizer: 第39回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜（神奈川県・横浜市）
  • Year and Date: 2016-11-30
• [Presentation] ALS2疾患原因変異体の高次構造および細胞内局在の変化はALS2関連運動ニューロン疾患の発症要因である (2016)
  • Author(s): 佐藤海、鈴木-宇都宮恭子、平塚結衣、三井駿、小野鈴花、大友麻子、秦野伸二
  • Organizer: 第39回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜（神奈川県・横浜市）
  • Year and Date: 2016-11-30
• [Presentation] Development of novel device with high-throughout axonal transport quantification. (2016)
  • Author(s): Yokoyama, S., Wada, J., Kushida, T., Araki, R., Ishida, T., Otomo, A., Hadano, S., and Kimura, H.
  • Organizer: The 20th International Conference on Miniaturized Systems for Chemistry and Life Sciences
  • Place of Presentation: Dublin (Republic of Ireland)
  • Year and Date: 2016-10-09
  • Int'l Joint Research
• [Presentation] 明暗条件がマウスの行動に及ぼす影響 (2016)
  • Author(s): 内藤佳津子、上條あけみ、岩尾佳代子、秦野伸二
  • Organizer: 第50回日本実験動物技術者協会総会
  • Place of Presentation: ウェスタ川越（埼玉県・川越市）
  • Year and Date: 2016-09-29
• [Presentation] A novel teaching approach based on a DNA experiment lesson in high school biology laboratory class: Scientific inquiry by visualization of DNA sequence conservation and diversity and a developmental lesson to understand genetic diagnosis. (2016)
  • Author(s): Inoue, Y., Otomo, A., Takahashi, K., Moriya, H., Ohnuki, Y., Izumi, S., Miyachi, H., and Hadano, S.
  • Organizer: 2016 International Conference of East-Asian Association for Science Education (EASE2016 Tokyo)
  • Place of Presentation: 東京理科大学（東京都・新宿区）
  • Year and Date: 2016-08-26
  • Int'l Joint Research
• [Presentation] Loss of SQSTM1/p62 but not NFE2L2/Nrf2 exacerbates motor neuron degeneration in a mutant SOD1-expressing mouse ALS model. (2016)
  • Author(s): Hadano, S., Mitsui, S., Otomo, A., Sato, K., Ono, S., Chen, X. P., Aoki, M., Warabi, E., Yamamoto, M., Ishii, T., Yanagawa, T., Shang, H. -F., and Yoshii, F.
  • Organizer: 15th Asian and Oceanian Congress of Neurology
  • Place of Presentation: Kuala Lumpur (Malaysia)
  • Year and Date: 2016-08-19
  • Int'l Joint Research
• [Presentation] Loss of SQSTM1/p62 but not NFE2L2/Nrf2 exacerbates motor neuron degeneration in a mutant SOD1-expressing mouse ALS model. (2016)
  • Author(s): Hadano, S., Mitsui, S., Otomo, A., Sato, K., Ono, S., Chen, X. P., Aoki, M., Warabi, E., Yamamoto, M., Ishii, T., Yanagawa, T., Shang, H. -F., and Yoshii, F.
  • Organizer: 15th Asian and Oceanian Congress of Neurology
  • Place of Presentation: Kuala Lumpur Convention Centre (Kuala Lumpur, Malaysia)
  • Year and Date: 2016-08-18
  • Int'l Joint Research
• [Presentation] Analysis of axonal transport in cultured neurons from ALS model mouse by using the microfluidic cell culture system. (2016)
  • Author(s): Otomo, A., Araki, R., Yokoyama, S., Wada, J., Hadano, S., and Kimura, H.
  • Organizer: 第39回日本神経科学大会（Neuroscience2016）
  • Place of Presentation: パシフィコ横浜（神奈川県・横浜市）
  • Year and Date: 2016-07-20
• [Presentation] The novel ALS2-interacting small G protein Rab17 colocalizes with ALS2/Alsin in recycling endosomes. (2016)
  • Author(s): Ono, S., Otomo, A., Fukuda, M., and Hadano, S.
  • Organizer: 第39回日本神経科学大会（Neuroscience2016）
  • Place of Presentation: パシフィコ横浜（神奈川県・横浜市）
  • Year and Date: 2016-07-20
• [Presentation] The novel ALS2-interacting small G protein Rab17 colocalizes with ALS2/Alsin in recycling endosomes. (2016)
  • Author(s): Ono, S., Otomo, A., Fukuda, M., and Hadano, S.
  • Organizer: 第39回日本神経科学大会
  • Place of Presentation: パシフィコ横浜（神奈川県、横浜市）
  • Year and Date: 2016-07-20
• [Presentation] Analysis of axonal transport in cultured neurons from ALS model mouse by using the microfluidic cell culture system. (2016)
  • Author(s): Otomo, A., Araki, R., Yokoyama, S., Wada, J., Hadano, S., and Kimura, H.
  • Organizer: 第39回日本神経科学大会
  • Place of Presentation: パシフィコ横浜（神奈川県、横浜市）
  • Year and Date: 2016-07-20
• [Presentation] マイクロ流体デバイスを用いた神経疾患モデルの確立. (2016)
  • Author(s): 荒木良介、大友麻子、横山奨、和田純希、秦野伸二、木村啓志
  • Organizer: 細胞アッセイ研究会
  • Place of Presentation: 東京大学生産技術研究所コンベンションホール（東京都、文京区）
  • Year and Date: 2016-01-19
• [Presentation] Molecular pathogenesis of ALS: Dysregulation of membrane trafficking and proteostasis. (2016)
  • Author(s): Hadano, S.
  • Organizer: Annual conference 2016: Movement Disorders and Motor Neuron Disease, Society of Neurology in Sichuan Province
  • Place of Presentation: Chengdu (China)
  • Int'l Joint Research / Invited
• [Presentation] 神経変性疾患の発症機序解明に向けた新たな展開 (2016)
  • Author(s): 秦野伸二
  • Organizer: 東海大学総合医学研究所第12回研修会
  • Place of Presentation: 東海大学医学部（神奈川県・伊勢原市）
• [Presentation] The novel ALS2-interacting small G protein Rab17 colocalizes with ALS2 in recycling endosomes. (2015)
  • Author(s): Ono, S., Otomo, A., Onodera, W., Sato, K., Mitsui, S., Fukuda, M., and Hadano, S.
  • Organizer: 26th International Symposium on ALS/MND
  • Place of Presentation: JM Marriott Orlando Grande Lakes (Orlando, U.S.A.)
  • Year and Date: 2015-12-11
  • Int'l Joint Research
• [Presentation] SOD1H46R発現ALSマウスモデルの運動ニューロン変性はNrf2ではなく、p62/SQSTM1の機能喪失により悪化する. (2015)
  • Author(s): 三井駿、久保瑞希、潘雷、大友麻子、小池正人、内山安男、青木正志、山本雅之、石井哲郎、柳川徹、Hui-Fang Shang、吉井文均、秦野伸二
  • Organizer: BMB2015・第38回日本分子生物学会年会・第88回日本生化学会大会
  • Place of Presentation: 神戸国際会議場（兵庫県、神戸市）
  • Year and Date: 2015-12-01
• [Presentation] ALS2疾患原因変異体はALS2複合体構造や細胞内局在を変化させる. (2015)
  • Author(s): 佐藤海、平塚結衣、大友麻子、秦野伸二
  • Organizer: BMB2015・第38回日本分子生物学会年会・第88回日本生化学会大会
  • Place of Presentation: 神戸国際会議場（兵庫県、神戸市）
  • Year and Date: 2015-12-01
• [Presentation] ALS2及び新規結合低分子量Gタンパク質Rab17はリサイクリングエンドソームに局在する. (2015)
  • Author(s): 小野鈴花、大友麻子、福田光則、秦野伸二
  • Organizer: BMB2015・第38回日本分子生物学会年会・第88回日本生化学会大会
  • Place of Presentation: 神戸国際会議場（兵庫県、神戸市）
  • Year and Date: 2015-12-01
• [Presentation] マイクロ流体デバイスを用いたALS疾患モデルの確立. (2015)
  • Author(s): 荒木良介、大友麻子、横山奨、和田純希、秦野伸二、木村啓志
  • Organizer: BMB2015・第38回日本分子生物学会年会・第88回日本生化学会大会
  • Place of Presentation: 神戸国際会議場（兵庫県、神戸市）
  • Year and Date: 2015-12-01
• [Presentation] SQSTM1/p62及びその変異体の神経細胞における局在解析. (2015)
  • Author(s): 白川涼平、濱祐太郎、大友麻子、秦野伸二
  • Organizer: BMB2015・第38回日本分子生物学会年会・第88回日本生化学会大会
  • Place of Presentation: 神戸国際会議場（兵庫県、神戸市）
  • Year and Date: 2015-12-01
• [Presentation] Loss of p62/SQSTM1 but not Nrf2 accelerates motor neuron degeneration in SOD1H46R transgenic mice. (2015)
  • Author(s): Mitsui, S., Kubo, M., Pan, L., Otomo, A., Koike, M., Uchiyama, Y., Aoki, M., Yamamoto, M., Ishii, T., Yanagawa, T., Shang, H. -F., Yoshii, F., and Hadano, S.
  • Organizer: 第38回日本神経科学大会
  • Place of Presentation: 神戸国際会議場（兵庫県、神戸市）
  • Year and Date: 2015-07-28
• [Presentation] Pathogenic mutations in ALS2 alter the oligomeric states of the ALS2 complex and its subcellular localization. (2015)
  • Author(s): Sato, K., Hiratsuka, Y., Suzuki-Utsunomiya, K., Otomo, A., and Hadano, S.
  • Organizer: 第38回日本神経科学大会
  • Place of Presentation: 神戸国際会議場（兵庫県、神戸市）
  • Year and Date: 2015-07-28
• [Presentation] Neurological diseases: Recent advances in studies on genes and their functions. (2015)
  • Author(s): Hadano, S.
  • Organizer: 2015 Tianfu International Stroke Conference
  • Place of Presentation: Holiday Inn Chengdu Century City- West Tower (Chengdu, China)
  • Year and Date: 2015-05-30
  • Int'l Joint Research / Invited
• [Presentation] Motor neuron diseases: Recent advances in studies on genes and their functions. (2015)
  • Author(s): Hadano, S.
  • Organizer: Society of Neurology in Sichuan Province
  • Place of Presentation: Holiday Inn Chengdu Century City- West Tower (Chengdu, China)
  • Year and Date: 2015-05-30
  • Int'l Joint Research / Invited
• [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] Generation and characterization of a novel transgenic mouse model for monitoring the autophagy-endolysosomal system. (2014)
  • Author(s): Hadano, S., Wang, T., Tanaka, M., Hayashi, H., Ogiwara, S., Okada, C., Itoh, M., Fukunishi, N., Iida, Y., Nakamura, A., Otomo, A., and Ohtsuka, M.
  • 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] Identification and characterization of a novel ALS2 interacting protein Rab30. (2014)
  • Author(s): Onodera, W., Otomo, A., Fukuda, M., and Hadano, S.
  • 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] Subcellular distribution of ALS2 and the novel ALS2-interacting small G protein Rab17. (2014)
  • Author(s): Ono, S., Otomo, A., Fukuda, M., and Hadano, S.
  • 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: 日本実験動物科学技術さっぽろ2014：第61回日本実験動物学会総会/第48回日本実験動物技術者協会総会
  • Place of Presentation: 札幌
  • Year and Date: 2014-05-15 – 2014-05-17