Microarray analysis in spinal cords of sporadic ALS patients with cell-type specific transcriptome

• Project/Area Number: 24591259
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Neurology
• Research Institution: Kyoto University
• Principal Investigator: YAMASHITA Hirofumi 京都大学, 医学(系)研究科(研究院), 助教 (60402913)
• Co-Investigator(Renkei-kenkyūsha): YAMANAKA Koji 名古屋大学, 環境医学研究所, 教授 (80446533)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2013: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2012: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: 筋萎縮性側索硬化症 / マイクロアレイ / アストロサイト / ミクログリア / トランスクリプトーム / 自然免疫 / 遺伝子 / グリア

Outline of Final Research Achievements

We employed unbiased screening to identify significantly misregulated 173 genes in sporadic ALS spinal cords with microarray. Similar experiments have been done before, therefore we went further by newly focusing on the cell-types that express misregulated genes. With cell-type specific transcriptome, we could analyze the microarray data from spinal cord samples composed of heterogenous cell-types in full detail. Nearly half of the misregulated genes in ALS spinal cords were expressesd abunduntly in microglia or astrocytes. Many of those genes were also changed in ALS mouse models. We confirmed that the prediction for the cell type abundantly expressing the genes were correct by the immunohistochemistry for CLEC7A, GPR65, NOX2, CHI3L1, Oncostatin M receptor, CEBPD, and calponin 3 with ALS mouse spinal cords.
We could present the useful methods for analyzing microarray data derived from spinal cord samples composed of heterogenous cell-types.

Research Products

• [Journal Article] A novel mutation in <i>glycyl-tRNA synthetase</i> caused Charcot-Marie-Tooth disease type 2D with facial and respiratory muscle involvement (2014)
  • Author(s): N. Kawakami, K. Komatsu, H. Yamashita, K. Uemura,N.Oka, H. Takashima, and R. Takahashi
  • Journal Title: Rinsho Shinkeigaku Volume: 54 Issue: 11 Pages: 911-915
  • DOI: 10.5692/clinicalneurol.54.911
  • NAID: 130004706051
  • ISSN: 0009-918X, 1882-0654
  • Peer Reviewed
• [Journal Article] Immunoreactivity of valosin-containing protein in sporadic amyotrophic lateral sclerosis and in a case of its novel mutant (2014)
  • Author(s): Ayaki T, Ito H, Fukushima H, Inoue T, Kondo T, Ikemoto A, Asano T, Shodai A, Fujita T, Fukui S, Morino H, Nakano S, Kusaka H, Yamashita H, Ihara M, Matsumoto R, Kawamata J, Urushitani M, Kawakami H, Takahashi R.
  • Journal Title: Acta Neuropathol Commun. Volume: 2 Issue: 1 Pages: 172-185
  • DOI: 10.1186/s40478-014-0172-0
  • Peer Reviewed / Open Access
• [Journal Article] Optineurin suppression causes neuronal cell death via NF-κB pathway. (2013)
  • Author(s): Akizuki M, Yamashita H, Uemura K, Maruyama H, Kawakami H, Ito H, Takahashi R.
  • Journal Title: J Neurochem Volume: 126(6) Issue: 6 Pages: 2230-41
  • DOI: 10.1111/jnc.12326
  • Peer Reviewed / Open Access
• [Presentation] Microarray analysis in spinal cords of sporadic ALS patients with cell-type specific transcriptome (2014)
  • Author(s): Hirofumi Yamashita, Okiru komine, Noriko Fujimori, Hidefumi Ito, Yohei Iguchi, Naoki Atsuta, Fumiaki Tanaka, Gen Sobue, Ryosuke Takahashi, Koji Yamanaka
  • Organizer: 第37回日本神経科学大会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2014-09-13
• [Presentation] Microarray analysis in spinal cords of sporadic ALS patients with cell-type specific transcriptome (2014)
  • Author(s): Hirofumi Yamashita 、 Noriko Fujimori 、 Hidefumi Ito 、Yohei Iguchi 、Naoki Atsuta 、 Fumiaki Tanaka 、Gen Sobue 、 Ryosuke Takahashi 、Koji Yamanaka
  • Organizer: Nucleic Acid Summit
  • Place of Presentation: San Diego,U.S.A.
  • Year and Date: 2014-06-19
• [Presentation] Optineurin suppression causes neuronal cell death via NF-κB pathway (2014)
  • Author(s): Akizuki M1, Yamashita H, Uemura K, Maruyama H, Kawakami H, Ito H, Takahashi R.
  • Organizer: 14th Asian & Oceanian Congress of Neurology
  • Place of Presentation: Macau
• [Presentation] NF-κB経路を介したOptineurinのノックダウンによる神経細胞死 (2013)
  • Author(s): 秋月真由美、山下博史、植村健吾、丸山博文、川上秀文、伊東秀文、髙橋良輔
  • Organizer: 第22回 日本Cell death学会
  • Place of Presentation: 京都 京都大学
• [Presentation] NF-κB経路を介したOptineurinのノックダウンによる神経細胞死 (2013)
  • Author(s): 秋月真由美、山下博史、植村健吾、丸山博文、川上秀文、伊東秀文、髙橋良輔
  • Organizer: Neuro2013
  • Place of Presentation: 京都 国際会議場
• [Presentation] Microarray analysis in spinal cords of sporadic ALS patients with cell-type specific transcriptome
  • Author(s): 山下博史
  • Organizer: The 64th Annual Meeting of the American Academy of Neurology
  • Place of Presentation: Morial Convention Cente, New Orleans, Louisiana, USA
• [Presentation] Microarray analysis in spinal cords of sporadic ALS patients with cell-type specific transcriptome
  • Author(s): 山下博史
  • Organizer: The 13th Asian Oceanian Congress of Neurology
  • Place of Presentation: Melbourne Convention and Exhibition Centre, Melbourne, Australia
• [Presentation] Microarray analysis in spinal cords of sporadic ALS patients with cell-type specific transcriptome
  • Author(s): 山下博史
  • Organizer: 23rd international symposium on ALS/MND
  • Place of Presentation: Sheraton Cicago Hotel & Towers, Cicago, Illinois, USA
• [Book] 分子脳科学 (2015)
  • Author(s): 山下博史・漆谷真・高橋良輔（三品 昌美 編）
  • Total Pages: 312
  • Publisher: 化学同人
• [Book] 医学のあゆみ(遺伝子改変によるALSモデルマウス) (2015)
  • Author(s): 山下博史、漆谷真
  • Publisher: 医歯薬出版
• [Book] medicina(筋萎縮～特徴的な所見と検査) (2014)
  • Author(s): 山下博史,髙橋良輔
  • Publisher: 医学書院
• [Book] 脳神経科学イラストレイテッド 改訂第3版 「ALSなど運動ニューロン病」の章 (2013)
  • Author(s): 山下博史、髙橋良輔
  • Total Pages: 395
  • Publisher: 羊土社
• [Book] 脳神経科学イラストレイテッド 改訂第3版 (2013)
  • Author(s): 山下博史、髙橋良輔
  • Publisher: 羊土社