The analyses of pathomechanisms of ALS and its development of therapeutic strategy by using Drosophila model.

• Project/Area Number: 16K19519
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Neurology
• Research Institution: Kyoto Prefectural University of Medicine
• Principal Investigator: AZUMA YUMIKO 京都府立医科大学, 医学(系)研究科(研究院), 特任助教 (00453093)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2017: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2016: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 筋萎縮性側索硬化症 / ショウジョウバエ / FUS / TDP-43 / FIG4 / 遺伝学 / 神経変性疾患 / TBPH / Cabeza / ALS / 脳神経疾患 / 応用生物学

Outline of Final Research Achievements

Amyotrophic lateral sclerosis (ALS) causes the progressive loss of motor neurons leading to muscle weakening and respiratory paralysis. To elucidate the common pathomechanisms of　neurodegeneration in ALS in vivo, we explored common modifier genes by using Drosophila FUS, TDP-43 and FIG4 knockdown. We identified autophagy-related genes VCP as modifiers of neurodegeneration in ALS model flies expressing TDP-43. Moreover, We identified MST as modifiers of neurodegeneration in ALS model flies expressing Drosophila FUS and Drosophila FIG4. MST is associated with the regulatory mechanisms for many biological events including cell death, cell proliferation, autophagy, and others. Our results suggest that both VCP and MST is involved in common pathogenesis of neurodegeneration in ALS.

Academic Significance and Societal Importance of the Research Achievements

ショウジョウバエは遺伝学・発生学のモデル生物の中でも最も多様な解析手法が確立しており、病態解明に大きな貢献を果たしてきた。model flyを用いた研究は、世代時間が短く、遺伝子組換え体の作成が簡便にでき、ヒト疾患関連遺伝子の約75%が相同遺伝子として存在し、多数の遺伝子変異体株がストックセンターから容易に入手することが出来る。このようなショウジョウバエの特性を最大限に活用して、複数の異なるALSの原因遺伝子を用いたモデルショウジョウバエを解析することにより、共通する分子メカニズムが明らかになれば、家族性および孤発性ALS全体の病態解明や根本治療の開発への応用が期待される。

Research Products

• [Journal Article] Hippo, Drosophila MST, is a novel modifier of motor neuron degeneration induced by knockdown of Caz, Drosophila FUS. (2018)
  • Author(s): Azuma Y, Tokuda T, Kushimura Y, Yamamoto I, Mizuta I, Mizuno T, Nakagawa M, Ueyama M, Nagai Y, Iwasaki Y, Yoshida M, Pan D, Yoshida H, Yamaguchi M.
  • Journal Title: Exp Cell Res. Volume: 371 Issue: 2 Pages: 311-321
  • DOI: 10.1016/j.yexcr.2018.08.001
  • Peer Reviewed / Open Access
• [Journal Article] Loss-of-function mutation in Hippo suppressed enlargement of lysosomes and neurodegeneration caused by dFIG4 knockdown. (2018)
  • Author(s): Kushimura, Y., Azuma, Y., Mizuta, I., Muraoka, Y., Kyotani, A., Yoshida, H., Tokuda, T., Mizuno, T., and Yamaguchi, M.
  • Journal Title: Neuroreport Volume: 29 Issue: 10 Pages: 856-862
  • DOI: 10.1097/wnr.0000000000001044
  • Peer Reviewed / Open Access
• [Journal Article] verexpression of ter94, Drosophila VCP, improves motor neuron degeneration induced by knockdown of TBPH, Drosophila TDP-43. (2018)
  • Author(s): Kushimura Y, Tokuda T, Azuma Y, Yamamoto I, Mizuta I, Mizuno T, Nakagawa M, Ueyama M, Nagai Y, Yoshida H, Yamaguchi M.
  • Journal Title: Am J Neurodegener Dis Volume: 7 Pages: 11-31
  • Peer Reviewed / Open Access
• [Journal Article] NPM-hMLF1 fusion protein suppresses defects of a Drosophila FTLD model expressing the human FUS gene. (2018)
  • Author(s): Yamamoto I, Azuma Y, Kushimura Y, Yoshida H, Mizuta I, Mizuno T, Ueyama M, Nagai Y, Tokuda T, Yamaguchi M.
  • Journal Title: Sci. Rep. Volume: 8 Issue: 1 Pages: 11291-11291
  • DOI: 10.1038/s41598-018-29716-9
  • Peer Reviewed / Open Access
• [Journal Article] Genetic screening of the genes interacting with Drosophila FIG4 identified a novel link between CMT-causing gene and long noncoding RNAs. (2018)
  • Author(s): Muraoka Y, Nakamura A, Tanaka R, Suda K, Azuma Y, Kushimura Y, Lo Piccolo L, Yoshida H, Mizuta I, Tokuda T, Mizuno T, Nakagawa M, Yamaguchi M.
  • Journal Title: Exp. Neurol. Volume: 310 Pages: 1-13
  • DOI: 10.1016/j.expneurol.2018.08.009
  • Peer Reviewed / Open Access
• [Journal Article] Novel Drosophila model for mitochondrial diseases by targeting of a solute carrier protein SLC25A46. (2018)
  • Author(s): Suda, K., Ueoka, I., Azuma, Y., Muraoka, Y., Yoshida, H. and Yamaguchi, M.
  • Journal Title: Brain Research Volume: 1689 Pages: 30-44
  • DOI: 10.1016/j.brainres.2018.03.028
  • Peer Reviewed / Open Access
• [Journal Article] Novel roles of Drosophila FUS and Aub responsible for piRNA biogenesis in neuronal disorders. (2018)
  • Author(s): Wakisaka KT, Tanaka R, Hirashima T, Muraoka Y, Azuma Y, Yoshida H, Tokuda T, Asada S, Suda K, Ichiyanagi K, Ohno S, Itoh M, Yamaguchi M.
  • Journal Title: Brain Res. Volume: 18 Pages: 30646-30652
  • DOI: 10.1016/j.brainres.2018.12.028
  • Peer Reviewed / Open Access
• [Journal Article] Overexpression of ter94, Drosophila VCP, improves motor neuron degeneration induced by knockdown of TBPH, Drosophila TDP-43. (2018)
  • Author(s): Kushimura Y, Tokuda T, Azuma Y, Yamamoto I, Mizuta I, Mizuno T, Nakagawa M, Ueyama M, Nagai Y, Yoshida H, Yamaguchi M.
  • Journal Title: Am J Neurodegener Dis Volume: 7 Pages: 11-31
  • Peer Reviewed / Open Access
• [Journal Article] ALS and cancer (2016)
  • Author(s): Yamaguchi M, Azuma Y and Yoshida H.
  • Journal Title: J Carcinog Mutagen Volume: 7 Issue: 06
  • DOI: 10.4172/2157-2518.1000e122
  • Peer Reviewed / Open Access
• [Presentation] The loss-of-function mutation of Hippo improves enlargement of lysosome and neurodegeneration caused by dFIG4 knockdown. (2018)
  • Author(s): Kushimura Y, Azuma Y, Mizuta I, Muraoka Y, Kyotani A, Yoshida H, Tokuda T, Mizuno T, Yamaguchi M.
  • Organizer: 16th Asian Oceanian Congress of Neurology.
• [Presentation] ショウジョウバエCMT原因遺伝子dFIG4と遺伝学的に相互作用する遺伝子の同定. (2018)
  • Author(s): 村岡夕香,中村綾,田中領,須田晃治郎,東裕美子,櫛村由紀恵, Luca Lo Piccolo,吉田英樹, 水田依久子,徳田隆彦,水野敏樹,山口政光.
  • Organizer: 第41回日本分子生物学会
• [Presentation] ALSモデルバエにおけるpiRNA産生に応答した，ショウジョウバエFUSとAubの新規の役割. (2018)
  • Author(s): 脇坂啓子, 田中領,平島智貴,村岡夕香,東裕美子, 吉田英樹, 徳田隆彦, 浅田賢志, 須田晃治郎, 伊藤雅信, 山口政光.
  • Organizer: 第41回日本分子生物学会
• [Presentation] The search for genes that modulate FUS-mediated phenotypes by Drosophila amyotrophic lateral sclerosis model. (2017)
  • Author(s): Azuma Y, Tokuda T, Kushimura Y, Yamamoto I, Ueoka I, Mizuta I, Mizuno T, Nakagawa M, Ueyama M, Nagai Y, Yoshida H, Yamaguchi M.
  • Organizer: Cold Spring Harbor Laboratory Meetings & Courses, Neurobiology of Drosophila.
  • Int'l Joint Research
• [Presentation] Loss of function mutation of ter94, Drosophila VCP, partially enhanced motor neuron degeneration induced by knockdown of TBPH, Drosophila TDP-43． (2017)
  • Author(s): Kushimura Y，Tokuda T，Azuma Y，Yamamoto I，Ueoka I，Mizuta I，Mizuno T，Nakagawa M，Ueyama M，Nagai Y，Yoshida H，Yamaguchi M．
  • Organizer: Cold Spring Harbor Laboratory Meetings & Courses, Neurobiology of Drosophila.
  • Int'l Joint Research
• [Presentation] The search for genes that modulate FUS-mediated phenotypes by Drosophila amyotrophic lateral sclerosis model. (2017)
  • Author(s): Azuma Y, Tokuda T, Kushimura Y, Yamamoto I, Ueoka I, Mizuta I, Mizuno T, Nakagawa M, Ueyama M, Nagai Y, Yoshida H, Yamaguchi M.
  • Organizer: XXⅢ World congress of neurology
  • Int'l Joint Research
• [Presentation] Loss of function mutation of ter94, Drosophila VCP, partially enhanced motor neuron degeneration induced by knockdown of TBPH, Drosophila TDP-43. (2017)
  • Author(s): Kushimura Y，Tokuda T，Azuma Y，Yamamoto I，Ueoka I，Mizuta I，Mizuno T，Nakagawa M，Ueyama M，Nagai Y，Yoshida H，Yamaguchi M．
  • Organizer: XXⅢ World congress of neurology
  • Int'l Joint Research
• [Presentation] The search for genes that modulate FUS-mediated phenotypes by Drosophila amyotrophic lateral sclerosis model. (2017)
  • Author(s): Azuma Y, Tokuda T, Kushimura Y, Yamamoto I, Ueoka I, Mizuta I, Mizuno T, Nakagawa M, Ueyama M, Nagai Y, Yoshida H, Yamaguchi M.
  • Organizer: 第40回日本分子生物学会
• [Presentation] Loss of function mutation of ter94, Drosophila VCP, partially enhanced motor neuron degeneration induced by knockdown of TBPH, Drosophila TDP-43． (2017)
  • Author(s): Kushimura Y，Tokuda T，Azuma Y，Yamamoto I，Ueoka I，Mizuta I，Mizuno T，Nakagawa M，Ueyama M，Nagai Y，Yoshida H，Yamaguchi M．
  • Organizer: 第40回日本分子生物学会
• [Presentation] ALS原因遺伝子FUSのショウジョウバエホモログCabezaと遺伝学的相互作用するエピジェネティック制御因子の同定 (2017)
  • Author(s): 浅田賢志, 鈴木孝禎, 東裕美子, 徳田隆彦, 吉田英樹, 山口政光
  • Organizer: 第40回日本分子生物学会
• [Presentation] ショウジョウバエモデルを用いたパーキンソン病（PD）原因候補遺伝子の解析 (2017)
  • Author(s): 清水丈, 笠井高士, 東裕美子, 吉田英樹, 山口政光
  • Organizer: 第40回日本分子生物学会
• [Presentation] ショウジョウバエCMT原因遺伝子dFIG4の新規遺伝学的相互作用因子としての長鎖非コードRNA, CR18854の同定 (2017)
  • Author(s): 村岡夕香, 中村綾, 田中領, 東裕美子, 櫛村由紀恵, Luca Lo Piccolo, 須田晃治郎, 吉田英樹, 水野敏樹, 徳田隆彦, 山口政光
  • Organizer: 第40回日本分子生物学会
• [Presentation] FUS regulates synapse structure and function at the neuromuscular junctions by Drosophila ALS model (2016)
  • Author(s): 東裕美子, 徳田隆彦, 櫛村由紀恵,山本格, 京谷茜, 水田依久子, 水野敏樹, 中川正法, 上山盛夫, 永井義隆, 吉田英樹, 山口政光
  • Organizer: 日本分子生物学会
  • Place of Presentation: 横浜
  • Year and Date: 2016-11-30
• [Presentation] The functional analysis of TDP-43 by Drosophila ALS model (2016)
  • Author(s): 櫛村由紀恵, 徳田隆彦, 東裕美子, 山本格, 京谷茜, 水田依久子, 水野敏樹, 中川正法, 上山盛夫, 永井義隆, 吉田英樹, 山口政光
  • Organizer: 日本分子生物学会
  • Place of Presentation: 横浜
  • Year and Date: 2016-11-30
• [Presentation] ヒトNPM-hMLF1融合タンパク質はALS原因因子FUSが誘導する凝集体形成を阻害する (2016)
  • Author(s): 山本格, 東裕美子, 櫛村由紀恵, 吉田英樹, 水田依久子, 奥主隆太, 上山盛夫, 藤掛伸宏, 水野敏樹, 徳田隆彦, 山口 政光
  • Organizer: 日本分子生物学会
  • Place of Presentation: 横浜
  • Year and Date: 2016-11-30
• [Presentation] ショウジョウバエCMT原因遺伝子／ALS関連遺伝子FIG4の遺伝学的相互作用因子としてのイオンチャンネル活性化因子とlong non-coding RNAの同定 (2016)
  • Author(s): 中村綾, 山本格, 吉田英樹, 東裕美子, 水田依久子, 水野敏樹, 中川正法, 徳田隆彦.山口政光.
  • Organizer: 日本分子生物学会
  • Place of Presentation: 横浜
  • Year and Date: 2016-11-30
• [Presentation] FUS regulates synapse structure and function at the neuromuscular junctions by Drosophila ALS model (2016)
  • Author(s): 東裕美子, 徳田隆彦, 櫛村由紀恵,山本格, 京谷茜, 水田依久子, 水野敏樹, 中川正法, 上山盛夫, 永井義隆, 吉田英樹, 山口政光
  • Organizer: 日本神経学会
  • Place of Presentation: 神戸
  • Year and Date: 2016-05-18
• [Presentation] The functional analysis of TDP-43 by Drosophila ALS model (2016)
  • Author(s): 櫛村由紀恵, 徳田隆彦, 東裕美子, 山本格, 京谷茜, 水田依久子, 水野敏樹, 中川正法, 上山盛夫, 永井義隆, 吉田英樹, 山口政光
  • Organizer: 日本神経学会
  • Place of Presentation: 神戸
  • Year and Date: 2016-05-18
• [Presentation] ヒトNPM-hMLF1融合タンパク質はALS原因因子FUSが誘導する凝集体形成を阻害する (2016)
  • Author(s): 山本格, 東裕美子, 櫛村由紀恵, 吉田英樹, 水田依久子, 奥主隆太, 上山盛夫, 藤掛伸宏, 水野敏樹, 徳田隆彦, 山口 政光
  • Organizer: 第6回４大学連携研究フォーラム
  • Place of Presentation: 京都
• [Book] Drosophila models of human diseases. Masamitsu Yamaguchi Editor (chapter 6 Amyotrophic Lateral Sclerosis model) (2018)
  • Author(s): Azuma Y, Mizuta I, Tokuda T, and Mizuno T.
  • Total Pages: 301
  • Publisher: Springer
  • ISBN: 9789811305283