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Functional-structural relationship of liquid-liquid phase separation caused by ALS-causing proteins and its pathological significance.

Research Project

Project/Area Number 20H03593
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 52020:Neurology-related
Research InstitutionTokyo Medical University

Principal Investigator

Kanekura Kohsuke  東京医科大学, 医学部, 准教授 (10508568)

Co-Investigator(Kenkyū-buntansha) 早水 裕平  東京工業大学, 物質理工学院, 准教授 (80443216)
Project Period (FY) 2020-04-01 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥17,810,000 (Direct Cost: ¥13,700,000、Indirect Cost: ¥4,110,000)
Fiscal Year 2022: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000)
Fiscal Year 2021: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2020: ¥7,280,000 (Direct Cost: ¥5,600,000、Indirect Cost: ¥1,680,000)
Keywords液液相分離 / 筋萎縮性側索硬化症 / C9ORF72 / TDP-43 / ALS / 非膜性オルガネラ / LLPS / C9orf72
Outline of Research at the Start

真核生物は細胞内の特定の部位において生体高分子のやわらかな凝集を作り、生命活動を効率よく進める。この凝集の形成過程に液液相分離(LLPS)と呼ばれる現象が関与することが明らかとなってきた。近年神経変性疾患の様々な原因蛋白がLLPSを起こすことが報告され、特に筋萎縮性側索硬化症(ALS)においてはC9orf72、FUS、TDP43など多数のRNA結合原因蛋白がLLPSを起こす。本研究はこれらLLPSを起こすALS原因蛋白に対して集中的な構造機能解析を行い、LLPS制御の分子機構を明らかにすることで、LLPS異常によるALS発症解明の端緒となる知見を得ることを目指す。

Outline of Final Research Achievements

ALS-related proteins such as C9ORF72 dipeptide repeat proteins and TDP-43 are thought to contribute to ALS pathogenesis by disturbing phase separation homeostasis, but their molecular mechanisms have remained unclear. In this study, we analyzed why C9ORF72-derived poly(PR) is toxic by combining various structural variants, multi-omics analyses, and in silico analysis, and found that although the insertion of Pro between Arg is disadvantageous in terms of binding energy, it promotes multivalent binding and enhanced phase separation, resulting in toxicity.

Academic Significance and Societal Importance of the Research Achievements

本研究の成果により、C9ORF72由来ジペプチドの機能構造連関が初めて明らかとなった。本研究を応用することで、C9ORF72-poly(PR)の毒性機構を理解し、さらに相分離異常を標的とした新規治療法の開発にもつながる可能性がある。また、本研究は非膜性オルガネラの形成機構など広く科学分野へ寄与する可能性がある。研究生はは論文の出版や学会発表を通じて学術界で知見を共有し、さらに一般社会へ還元される。

Report

(4 results)
  • 2022 Annual Research Report   Final Research Report ( PDF )
  • 2021 Annual Research Report
  • 2020 Annual Research Report
  • Research Products

    (27 results)

All 2023 2022 2021 2020 Other

All Journal Article (18 results) (of which Int'l Joint Research: 5 results,  Peer Reviewed: 18 results,  Open Access: 16 results) Presentation (7 results) (of which Invited: 1 results) Remarks (2 results)

  • [Journal Article] How can we interpret the relationship between liquid-liquid phase separation and amyotrophic lateral sclerosis?2022

    • Author(s)
      Kanekura Kohsuke、Kuroda Masahiko
    • Journal Title

      Laboratory Investigation

      Volume: 102 Issue: 9 Pages: 912-918

    • DOI

      10.1038/s41374-022-00791-x

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Nuclear microRNAs release paused Pol II via the DDX21-CDK9 complex2022

    • Author(s)
      Shin-Ichiro Ohno, Keiki Oikawa, Toshiaki Tsurui, Yuichirou Harada, Kana Ono, Mizumo Tateishi, Aashiq Mirza, Masakatsu Takanashi, Kosuke Kanekura, Kumiko Nagase, Yoshihisa Shimada, Yujin Kudo, Norihiko Ikeda, Takahiro Ochiya, Xiaozhong Wang , Masahiko Kuroda
    • Journal Title

      Cell Reports

      Volume: 12;39(2) Issue: 2 Pages: 110673-110673

    • DOI

      10.1016/j.celrep.2022.110673

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Chemically oligomerizable TDP-43: a novel chemogenetic tool for studying the pathophysiology of amyotrophic lateral sclerosis2022

    • Author(s)
      Kanekura Kohsuke、Yamanaka Yoshiaki、Miyagi Tamami、Kuroda Masahiko
    • Journal Title

      Neural Regeneration Research

      Volume: 17 Issue: 11 Pages: 2434-2434

    • DOI

      10.4103/1673-5374.335803

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Effect of Multivalency on Phase-Separated Droplets Consisting of Poly(PR) Dipeptide Repeats and RNA at the Solid/Liquid Interface2022

    • Author(s)
      Chen Chen、Jia Han、Nakamura Yoshiki、Kanekura Kohsuke、Hayamizu Yuhei
    • Journal Title

      ACS Omega

      Volume: 7 Issue: 23 Pages: 19280-19287

    • DOI

      10.1021/acsomega.2c00811

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] The Patterning and Proportion of Charged Residues in the Arginine-Rich Mixed-Charge Domain Determine the Membrane-Less Organelle Targeted by the Protein2022

    • Author(s)
      Miyagi Tamami、Yamazaki Rio、Ueda Koji、Narumi Satoshi、Hayamizu Yuhei、Uji-i Hiroshi、Kuroda Masahiko、Kanekura Kohsuke
    • Journal Title

      International Journal of Molecular Sciences

      Volume: 23 Issue: 14 Pages: 7658-7658

    • DOI

      10.3390/ijms23147658

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Order controls disordered droplets: structure-function relationships in C9ORF72-derived poly(PR)2022

    • Author(s)
      Kanekura Kohsuke、Hayamizu Yuhei、Kuroda Masahiko
    • Journal Title

      American Journal of Physiology-Cell Physiology

      Volume: 322 Issue: 2 Pages: C197-C204

    • DOI

      10.1152/ajpcell.00372.2021

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] An improved macromolecular crowding sensor CRONOS for detection of crowding changes in membrane-less organelles under stressed conditions.2021

    • Author(s)
      Miyagi T, Yamanaka Y, Harada Y, Narumi S, Hayamizu Y, Kuroda M, Kanekura K.
    • Journal Title

      Biochem Biophys Res Commun.

      Volume: 583 Pages: 29-34

    • DOI

      10.1016/j.bbrc.2021.10.055

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Phase separation and toxicity of C9orf72 poly(PR) depends on alternate distribution of arginine.2021

    • Author(s)
      Chen C, Yamanaka Y, Ueda K, Li P, Miyagi T, Harada Y, Tezuka S, Narumi S, Sugimoto M, Kuroda M, Hayamizu Y, Kanekura K.
    • Journal Title

      J Cell Biol.

      Volume: 220 Issue: 11

    • DOI

      10.1083/jcb.202103160

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Expression of L-type amino acid transporter 1 is a poor prognostic factor for Non-Hodgkin's lymphoma.2021

    • Author(s)
      Jigjidkhorloo N, Kanekura K, Matsubayashi J, Akahane D, Fujita K, Oikawa K, Kurata A, Takanashi M, Endou H, Nagao T, Gotoh A, Norov O, Kuroda M.
    • Journal Title

      Sci Rep.

      Volume: 11(1) Issue: 1 Pages: 21638-21638

    • DOI

      10.1038/s41598-021-00811-8

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Establishment of chemically oligomerizable TAR DNA-binding protein-43 which mimics amyotrophic lateral sclerosis pathology in mammalian cells2021

    • Author(s)
      Yamanaka Yoshiaki、Miyagi Tamami、Harada Yuichiro、Kuroda Masahiko、Kanekura Kohsuke
    • Journal Title

      Laboratory Investigation

      Volume: 101 Issue: 10 Pages: 1331-1340

    • DOI

      10.1038/s41374-021-00623-4

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Diffusion of LLPS Droplets Consisting of Poly(PR) Dipeptide Repeats and RNA on Chemically Modified Glass Surface2021

    • Author(s)
      Chen Chen、Li Peiying、Luo Wei、Nakamura Yoshiki、Dimo Vanessa Seudo、Kanekura Kohsuke、Hayamizu Yuhei
    • Journal Title

      Langmuir

      Volume: 37 Issue: 18 Pages: 5635-5641

    • DOI

      10.1021/acs.langmuir.1c00493

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Acerola exosome-like nanovesicles to systemically deliver nucleic acid medicine via oral administration.2021

    • Author(s)
      Umezu T, Takanashi M, Murakami Y, Ohno SI, Kanekura K, Sudo K, Nagamine K, Takeuchi S, Ochiya T, Kuroda M.
    • Journal Title

      Molecular therapy. Methods & clinical development.

      Volume: 10;21 Pages: 199-208

    • DOI

      10.1016/j.omtm.2021.03.006

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Neuroplastin Modulates Anti-inflammatory Effects of MANF2020

    • Author(s)
      Yagi Takuya、Asada Rie、Kanekura Kohsuke、Eesmaa Ave、Lindahl Maria、Saarma Mart、Urano Fumihiko
    • Journal Title

      iScience

      Volume: 23 Issue: 12 Pages: 101810-101810

    • DOI

      10.1016/j.isci.2020.101810

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Comprehensive analysis of liver and blood miRNA in precancerous conditions2020

    • Author(s)
      Umezu Tomohiro、Tsuneyama Koichi、Kanekura Kohsuke、Hayakawa Michiyo、Tanahashi Toshihito、Kawano Mitsuoki、Taguchi Y-h、Toyoda Hidenori、Tamori Akihiro、Kuroda Masahiko、Murakami Yoshiki
    • Journal Title

      Scientific Reports

      Volume: 10 Issue: 1 Pages: 21766-21766

    • DOI

      10.1038/s41598-020-78500-1

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Pathological processes in aqueous humor due to iris atrophy predispose to early corneal graft failure in humans and mice2020

    • Author(s)
      Yamaguchi Takefumi、Higa Kazunari、Yagi-Yaguchi Yukari、Ueda Koji、Noma Hisashi、Shibata Shinsuke、Nagai Toshihiro、Tomida Daisuke、Yasu-Mimura Ririko、Ibrahim Osama、Matoba Ryo、Tsubota Kazuo、Hamrah Pedram、Yamada Jun、Kanekura Kohsuke、Shimazaki Jun
    • Journal Title

      Science Advances

      Volume: 6 Issue: 20 Pages: 5195-5195

    • DOI

      10.1126/sciadv.aaz5195

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] A Novel Homozygous Mutation of Thyroid Peroxidase Gene Abolishes a Disulfide Bond Leading to Congenital Hypothyroidism2020

    • Author(s)
      Yakou Fumiyoshi、Suwanai Hirotsugu、Ishikawa Takuya、Itou Mariko、Shikuma Jumpei、Miwa Takashi、Sakai Hiroyuki、Kanekura Kohsuke、Narumi Satoshi、Suzuki Ryo、Odawara Masato
    • Journal Title

      International Journal of Endocrinology

      Volume: 2020 Pages: 1-8

    • DOI

      10.1155/2020/9132372

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] A soluble endoplasmic reticulum factor as regenerative therapy for Wolfram syndrome2020

    • Author(s)
      Mahadevan Jana、Morikawa Shuntaro、Yagi Takuya、Abreu Damien、Lu Simin、Kanekura Kohsuke、Brown Cris M.、Urano Fumihiko
    • Journal Title

      Laboratory Investigation

      Volume: 100 Issue: 9 Pages: 1197-1207

    • DOI

      10.1038/s41374-020-0436-1

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] miR-27a ameliorates chemoresistance of breast cancer cells by disruption of reactive oxygen species homeostasis and impairment of autophagy2020

    • Author(s)
      Ueda S, Takanashi M, Sudo K, Kanekura K, Kuroda M
    • Journal Title

      Lab Invest.

      Volume: - Issue: 6 Pages: 863-873

    • DOI

      10.1038/s41374-020-0409-4

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Presentation] 筋萎縮性側索硬化症(ALS)原因ジペプチドリピートの細胞内局在および毒性メカニズムの解明.2023

    • Author(s)
      宮城碧水、金蔵孝介
    • Organizer
      第147回日本薬理学会関東部会
    • Related Report
      2022 Annual Research Report
  • [Presentation] アミノ酸の性質から紐解くALS原因ジペプチドリピートの相分離と毒性機構2022

    • Author(s)
      金蔵孝介
    • Organizer
      第16回日本アミノ酸学会
    • Related Report
      2022 Annual Research Report
  • [Presentation] Alternating Arg distribution controls phase separation and toxicity of poly(PR).2022

    • Author(s)
      金蔵孝介
    • Organizer
      Neuro2022
    • Related Report
      2022 Annual Research Report
  • [Presentation] Alternating Arg distribution controls phase separation and toxicity of poly(PR).2021

    • Author(s)
      金蔵孝介
    • Organizer
      第44回日本分子生物学会
    • Related Report
      2021 Annual Research Report
    • Invited
  • [Presentation] The alternating structure controls phase separation and toxicity of poly-PR C9orf72 dipeptides.2020

    • Author(s)
      金蔵孝介
    • Organizer
      日本生化学会
    • Related Report
      2020 Annual Research Report
  • [Presentation] C9orf72遺伝子産物poly-PRの構造機能連関と毒性獲得機構の解明2020

    • Author(s)
      金蔵孝介
    • Organizer
      日本認知症学会
    • Related Report
      2020 Annual Research Report
  • [Presentation] Molecular interactions of poly(PR) dipeptides in liquid liquid phase separation.2020

    • Author(s)
      Chen Chen, Kohsuke Kanekura, Yuhei Hayamizu
    • Organizer
      応用物理学会
    • Related Report
      2020 Annual Research Report
  • [Remarks] 東京医科大学分子病理学分野

    • URL

      https://tmumolpathol.sakura.ne.jp

    • Related Report
      2022 Annual Research Report
  • [Remarks] Research Map

    • URL

      https://researchmap.jp/10508568

    • Related Report
      2021 Annual Research Report

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Published: 2020-04-28   Modified: 2024-01-30  

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