Analyses for molecular chaperones regulating liquid-liquid phase separation in amyotrophic lateral sclerosis

• Project/Area Number: 20K16583
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 52020:Neurology-related
• Research Institution: Nara Medical University
• Principal Investigator: Nanaura Hitoki 奈良県立医科大学, 医学部, 助教 (00827909)
• Project Period (FY): 2020-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2021: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2020: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: 筋萎縮性側索硬化症 / 前頭側頭型認知症 / 液-液相分離 / シャペロン / アミロイド / 神経変性疾患

Outline of Research at the Start

筋萎縮性側索硬化症（ALS）をはじめとする神経変性疾患は、細胞内に蛋白凝集体を呈する疾患であるが、詳細な病態発症機序が未解明のため、有効な治療法はない。近年、ALS や関連疾患に、液-液相分離の制御異常が関連していることが明らかとなってきた。FUSやhnRNPA2などのRNA結合蛋白は液-液相分離するが、これらに生じた遺伝子変異がALSの原因となることが報告されている。本研究では、液-液相分離する蛋白質、および制御機能をもつシャペロンに関して、分野横断的解析を行うことにより、ALSをはじめとする神経変性疾患の病態発症機序を解明し、治療法の開発に繋げることを目指す。

Outline of Final Research Achievements

Aggregation and mislocalization of RNA binding proteins (RBPs) are observed in amyotrophic lateral sclerosis and other neurodegenerative diseases. Regulating self-association of RBPs to suppress the formation of pathogenic fibrils is crucial in the prevention of neurodegenerative diseases. Nuclear import receptors (NIRs) karyopherin-β2 (Kapβ2) not only controls the nucleocytoplasmic distribution of RBPs but also acts as a phase modifier to regulate self-association of FUS by recognizing proline–tyrosine nuclear localization signal. Exploiting multiple biochemical and biophysical techniques, we revealed in this study that arginine-rich poly-dipeptides from repeat expansion in C9orf72 impede the ability of NIRs to modify phase transitions of RBPs.

Academic Significance and Societal Importance of the Research Achievements

本研究により、毒性ペプチドが、相分離制御シャペロンの機能を阻害する詳細な分子メカニズムが明らかとなった。相分離の制御とその破綻に着目した本研究成果から、ALSをはじめとする神経変性疾患の病態解明、新たな治療法開発につながることが期待される。

Research Products

• [Journal Article] Serum Alkaline Phosphatase in Cryptogenic Stroke Cases with Active Cancer (2022)
  • Author(s): Izumi Tesseki、Nanaura Hitoki、Iguchi Naohiko、Ozaki Maki、Sugie Kazuma
  • Journal Title: Internal Medicine Volume: 61 Issue: 5 Pages: 639-646
  • DOI: 10.2169/internalmedicine.7550-21
  • NAID: 130008165973
  • ISSN: 0918-2918, 1349-7235
  • Year and Date: 2022-03-01
  • Peer Reviewed / Open Access
• [Journal Article] C9orf72-Derived Proline:Arginine Poly-Dipeptides Modulate Cytoskeleton and Mechanical Stress Response (2022)
  • Author(s): Shiota Tomo、Nagata Riko、Kikuchi Sotaro、Nanaura Hitoki、Matsubayashi Masaya、Nakanishi Mari、Kobashigawa Shinko、Isozumi Noriyoshi、Kiriyama Takao、Nagayama Kazuaki、Sugie Kazuma、Yamashiro Yoshito、Mori Eiichiro
  • Journal Title: Frontiers in Cell and Developmental Biology Volume: 10 Pages: 750829-750829
  • DOI: 10.3389/fcell.2022.750829
  • Peer Reviewed / Open Access
• [Journal Article] C9orf72-derived arginine-rich poly-dipeptides impede phase modifiers (2021)
  • Author(s): Nanaura Hitoki、Aiba Yuichiro、Nakanishi Mari、Shiota Tomo、Hibino Masaki、Kikuchi Sotaro、Nagata Riko、Morikawa Naritaka、Iguchi Naohiko、Kiriyama Takao、Nagamori Shushi、Sugie Kazuma、Saio Tomohide、Yoshizawa Takuya、Mori Eiichiro et al.
  • Journal Title: Nature Communications Volume: 12 Issue: 1 Pages: 5301-5301
  • DOI: 10.1038/s41467-021-25560-0
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Anti-programmed cell death-1 (PD-1) monoclonal antibodies involve reversible cranial dura matter (2021)
  • Author(s): Kataoka Hiroshi、Shimada Daisuke、Nanaura Hitoki、Sugie Kazuma
  • Journal Title: Oxford Medical Case Reports Volume: 2021 Issue: 9
  • DOI: 10.1093/omcr/omab077
  • Peer Reviewed / Open Access
• [Journal Article] Zonisamide for the Efficacy of Sleep Abnormality in Parkinson's Disease (ZEAL Study): A Protocol for Randomized Controlled Trials (2021)
  • Author(s): Kataoka Hiroshi、Isogawa Masahiro、Inoue Takashi、Hasebe Miyoko、Takashima Ryuzo、Kasama Shu、Nanaura Hitoki、Kiriyama Takao、Kasahara Masato、Sugie Kazuma
  • Journal Title: Frontiers in Neurology Volume: 12
  • DOI: 10.3389/fneur.2021.741307
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] 27‐Hydroxycholesterol regulates human SLC22A12 gene expression through estrogen receptor action (2020)
  • Author(s): Matsubayashi M、Sakaguchi YM.、Sahara Y、Nanaura H、Kikuchi S、Asghari A、Bui L、Kobashigawa S、Nakanishi M、Nagata R、Matsui TK.、Kashino G、Hasegawa M、Takasawa S、Eriguchi M、Tsuruya K、Nagamori S、Sugie K、Nakagawa T、Takasato M、Umetani M、Mori E
  • Journal Title: The FASEB Journal Volume: 35 Issue: 1
  • DOI: 10.1096/fj.202002077r
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Proteolytic cleavage of Podocin by Matriptase exacerbates podocyte injury (2020)
  • Author(s): Ozawa Shota、Matsubayashi Masaya、Nanaura Hitoki、Yanagita Motoko、Mori Kiyoshi、Asanuma Katsuhiko、Kajiwara Nobuyuki、Hayashi Kazuyuki、Ohashi Hiroshi、Kasahara Masato、Yokoi Hideki、Kataoka Hiroaki、Mori Eiichiro、Nakagawa Takahiko
  • Journal Title: Journal of Biological Chemistry Volume: 295 Issue: 47 Pages: 16002-16012
  • DOI: 10.1074/jbc.ra120.013721
  • Peer Reviewed / Open Access
• [Journal Article] Inhibition of the ATR kinase enhances 5-FU sensitivity independently of nonhomologous end-joining and homologous recombination repair pathways (2020)
  • Author(s): Ito Soichiro S.、Nakagawa Yosuke、Matsubayashi Masaya、Sakaguchi Yoshihiko M.、Kobashigawa Shinko、Matsui Takeshi K.、Nanaura Hitoki、Nakanishi Mari、Kitayoshi Fumika、Kikuchi Sotaro、Kajihara Atsuhisa、Tamaki Shigehiro、Sugie Kazuma、Kashino Genro、Takahashi Akihisa、Hasegawa Masatoshi、Mori Eiichiro、Kirita Tadaaki
  • Journal Title: Journal of Biological Chemistry Volume: 295 Issue: 37 Pages: 12946-12961
  • DOI: 10.1074/jbc.ra120.013726
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Brainstem organoids from human pluripotent stem cells (2020)
  • Author(s): Nobuyuki Eura, Takeshi K. Matsui, Joachim Luginbuhl, Masaya Matsubayashi, Hitoki Nanaura, Tomo Shiota, Kaoru Kinugawa, Naohiko Iguchi, Takao Kiriyama, Canbin Zheng, Tsukasa Kouno, Yan Jun Lan, Pornparn Kongpracha, Pattama Wiriyasermkul, Yoshihiko M. Sakaguchi, Riko Nagata, Tomoya Komeda, Naritaka Morikawa, et al.
  • Journal Title: Frontiers in Neuroscience Volume: - Pages: 00538-00538
  • DOI: 10.3389/fnins.2020.00538
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Dysfunction of molecular chaperones for phase separating proteins in ALS (2021)
  • Author(s): 七浦仁紀、塩田智、森英一朗、杉江和馬
  • Organizer: 第62回 日本神経学会学術大会
• [Presentation] C9orf72-derived PR poly-dipeptides target Kapβ2 through NLS binding site and dysregulate LLPS (2020)
  • Author(s): 七浦 仁紀, 塩田 智, 森 英一朗, 杉江 和馬
  • Organizer: 第61回日本神経学会学術大会