Elucidation of the pathogenesis of spinal and bulbar muscular atrophy by single cell RNA-seq analysis

• Project/Area Number: 21K20686
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Multi-year Fund
• Review Section: 0704:Neuroscience, brain sciences, and related fields
• Research Institution: Nagoya University
• Principal Investigator: Iida Madoka 名古屋大学, 医学系研究科, 助教 (40815437)
• Project Period (FY): 2021-08-30 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000); Fiscal Year 2022: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2021: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 球脊髄性筋萎縮症 / シングルセル解析 / オリゴデンドロサイト / イオンチャネル / シナプス / 興奮毒性 / 超早期病態 / 運動ニューロン疾患

Outline of Research at the Start

本研究課題では、シングルセル解析を用いてSBMAマウスモデルの脊髄における1細胞ごとの遺伝子発現情報とオープンクロマチン領域の分布を発症前から経時的に取得し、SBMAの早期病態解明と治療開発を目指す。運動ニューロンの遺伝子変化を効率的に検出するために、運動ニューロン特異的なレポーターを有するSBMAマウスモデルを用いる。シングルセル解析の結果に基づき、主に発症前に変化が大きい遺伝子と細胞種に注目し、神経系細胞の初代培養およびiPS細胞を用いた検証を行う。さらにSBMAマウスモデルを用いて、病期に応じて標的遺伝子が病態に与える影響を明らかにする。

Outline of Final Research Achievements

Spinal and bulbar muscular atrophy (SBMA) is a progressive hereditary neuromuscular disease caused by the expansion of CAG repeat in androgen receptor (AR). We aimed to elucidate the early pathogenesis of SBMA using single cell analysis. We obtained gene expression profile of each cell type in spinal cords of mouse model of SBMA (AR-97Q) from four different disease stages (3, 6, 9, and 13 weeks of age). Comparing wild-type mice with AR-97Q mice at each week, we found significant changes in oligodendrocytes, from 3 weeks: before onset of disease. It is also revealed that genes related to synapse and ion channel activity were up-regulated in oligodendrocytes of AR mice, suggesting that cellular excitotoxicity is involved in the pathogenesis of early stage of SBMA and that it could be a therapeutic target for SBMA.

Academic Significance and Societal Importance of the Research Achievements

球脊髄性筋萎縮症（SBMA）はそのアンドロゲン依存性病態に基づき、治療薬として抗アンドロゲン療法であるリュープロレリン酢酸塩が2017年に薬事承認されたが臨床における効果は限定的である。また性機能抑制などの副作用から、発症前のSBMAキャリア男性への使用は困難であり、新たな治療薬開発が望まれている。本研究によりSBMAの発症前にニューロンよりもオリゴデンドロサイトにおいて遺伝子発現変化が大きいことが判明し、従来は知られていなかった病態が明らかとなった。オリゴデンドロサイトの異常が運動ニューロンの変性の起始に強く関わっていると考えられ、疾患に対する新たな治療アプローチを提案することができた。

Research Products

• [Journal Article] Dysregulation of Aldh1a2 underlies motor neuron degeneration in spinal muscular atrophy (2023)
  • Author(s): Kataoka Mayumi、Sahashi Kentaro、Tsujikawa Koyo、Takeda Jun-ichi、Hirunagi Tomoki、Iida Madoka、Katsunoa Masahisa
  • Journal Title: Neuroscience Research Volume: - Pages: 58-65
  • DOI: 10.1016/j.neures.2023.04.007
  • Peer Reviewed / Open Access
• [Journal Article] Clinical implication of denervation in sporadic inclusion body myositis (2022)
  • Author(s): Noda Seiya、Murakami Ayuka、Kazuta Tomoyuki、Hirano Satoko、Kimura Seigo、Nakanishi Hirotaka、Matsuo Koji、Tsujikawa Koyo、Yamada Shinichiro、Iida Madoka、Koike Haruki、Kuru Satoshi、Katsuno Masahisa
  • Journal Title: Journal of the Neurological Sciences Volume: 439 Pages: 120317-120317
  • DOI: 10.1016/j.jns.2022.120317
  • Peer Reviewed / Open Access
• [Journal Article] Label-free morphological sub-population cytometry for sensitive phenotypic screening of heterogenous neural disease model cells (2022)
  • Author(s): Imai Yuta、Iida Madoka、Kanie Kei、Katsuno Masahisa、Kato Ryuji
  • Journal Title: Scientific Reports Volume: 12 Issue: 1 Pages: 9296-9296
  • DOI: 10.1038/s41598-022-12250-0
  • Peer Reviewed / Open Access
• [Journal Article] Mid1 is associated with androgen-dependent axonal vulnerability of motor neurons in spinal and bulbar muscular atrophy (2022)
  • Author(s): Ogura Yosuke、Sahashi Kentaro、Hirunagi Tomoki、Iida Madoka、Miyata Takaki、Katsuno Masahisa
  • Journal Title: Cell Death & Disease Volume: 13 Issue: 7 Pages: 601-601
  • DOI: 10.1038/s41419-022-05001-6
  • Peer Reviewed / Open Access
• [Journal Article] Metabolome and transcriptome analysis on muscle of sporadic inclusion body myositis. (2022)
  • Author(s): Murakami, A., Noda, S., Kazuta, T., Hirano, S., Kimura, S., Nakanishi, H., Matsuo, K., Tsujikawa, K., Iida, M., Koike, H., Sakamoto, K., Hara, Y., Kuru, S., Kadomatsu, K., Shimamura, T., Ogi, T., Katsuno, M.
  • Journal Title: Ann Clin Transl Neurol. Volume: 9 Issue: 10 Pages: 1602-1615
  • DOI: 10.1002/acn3.51657
  • Peer Reviewed / Open Access
• [Journal Article] Selective suppression of polyglutamine-expanded protein by lipid nanoparticle-delivered siRNA targeting CAG expansions in the mouse CNS. (2021)
  • Author(s): Hirunagi T, Sahashi K, Tachikawa K, Leu AI, Nguyen M, Mukthavaram R, Karmali PP, Chivukula P, Tohnai G, Iida M, Onodera K, Ohyama M, Okada Y, Okano H, Katsuno M.
  • Journal Title: Mol Ther Nucleic Acids. Volume: 24 Pages: 1-10
  • DOI: 10.1016/j.omtn.2021.02.007
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Ratio of urinary N-terminal titin fragment to urinary creatinine is a novel biomarker for amyotrophic lateral sclerosis (2021)
  • Author(s): Yamada Shinichiro、Hashizume Atsushi、Hijikata Yasuhiro、Ito Daisuke、Kishimoto Yoshiyuki、Iida Madoka、Koike Haruki、Hirakawa Akihiro、Katsuno Masahisa
  • Journal Title: Journal of Neurology, Neurosurgery & Psychiatry Volume: 92 Issue: 10 Pages: 1072-1079
  • DOI: 10.1136/jnnp-2020-324615
  • Peer Reviewed / Open Access
• [Journal Article] A role for the endocannabinoid enzymes monoacylglycerol and diacylglycerol lipases in cue-induced cocaine craving following prolonged abstinence. (2021)
  • Author(s): Mitra S, Gobira PH, Werner CT, Martin JA, Iida M, Thomas SA, Erias K, Miracle S, Lafargue C, An C, Dietz DM.
  • Journal Title: Addict Biol. Volume: 26 Issue: 5
  • DOI: 10.1111/adb.13007
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Single cell analysis reveals oligodendrocyte heterogeneity in spinal and bulbar muscular atrophy (2023)
  • Author(s): Madoka Iida
  • Organizer: 第64回日本神経学会学術大会
• [Presentation] Single cell analysis reveals oligodendrocyte heterogeneity in spinal and bulbar muscular atrophy (2023)
  • Author(s): Madoka Iida
  • Organizer: CAG Triplet Repeat Disorders Gordon Research Conference
  • Int'l Joint Research