細胞環境下での核膜孔の安定かつ過渡的なナノ構造測定技術を開発

• Project Area: Novel measurement techniques for visualizing 'live' protein molecules at work
• Project/Area Number: 17H05874
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Complex systems
• Research Institution: Kanazawa University
• Principal Investigator: WONG W・R 金沢大学, ナノ生命科学研究所, 教授 (30464035)
• Project Period (FY): 2017-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000); Fiscal Year 2018: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000); Fiscal Year 2017: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
• Keywords: 核膜孔 / ナノ構造 / 測定技術

Outline of Annual Research Achievements

我々の体の設計図であるDNAを格納する細胞核は、核膜と呼ばれる二重膜に覆われている。核内外の物質の能動的なやりとりには、核膜にある唯一のゲートである核膜孔が使用されている。核膜孔は核膜孔複合体（Nuclear Pore Complex: NPC）から成り立っており、その複合体は約30種類のNPCタンパク質（ヌクレオポリン）により構成されている。
本研究課題では、「細胞環境下での核膜孔の安定かつ過渡的なナノ構造測定技術を開発」に関する探索を行った。腫瘍のグレードと核構造の異常性とに相関があるため、核の形態はがん診断において重要な病理学的ターゲットとなり得る。ゆえに、我々は正常細胞とがん細胞におけるNPCのナノスケール構造の安定的かつ過渡的な変化や差異の詳細を解明するため、ナノ構造測定技術を開発することを目的とした。実施期間中、当研究課題の基礎となる、大腸がん細胞における核膜孔複合体のチャネル部分を構成していると報告されているタンパク質群、FG-Nups(フェニルアラニンーグリシンーヌクレオポリン)について構造およびダイナミクスのナノスケールにおける可視化を行った。これらの結果をまとめて論文とし、現在雑誌に投稿中である。
また、ヌクレオポリンの一種であるPOM121が、核内輸送タンパク質Importin-betaと共に、前立腺がんの治療において標的となり得ることをレビューにて発表した（Cell Chem. Biol., 2018 IF5.592）。
さらに、独自のヌクレオポリンを持つインフルエンザウイルスにおいて、ヘマグルチニン(HA)前駆体タンパク質そのものと、その構造変化のナノスケールにおける可視化に成功した（BBA-general Subjects, 2019 IF4.81）。

Research Progress Status

平成30年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

平成30年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Direct Visualization of Avian Influenza H5N1 Hemagglutinin Precursor and its Conformational Change by High-Speed Atomic Force Microscopy (2019)
  • Author(s): Lim SK, Mohamed MS, Wang H, Hartono, Hazawa M, Kobayashi A, Voon DC, Kodera N, Ando T and Wong RW
  • Journal Title: BBA - General Subjects Volume: 印刷中 Issue: 2 Pages: 129313-129313
  • DOI: 10.1016/j.bbagen.2019.02.015
  • Peer Reviewed / Open Access
• [Journal Article] Targeting nucleoporin POM121-importin β axis in prostate cancer. (2018)
  • Author(s): Lim SK and Wong RW
  • Journal Title: Cell Chem. Biol. Volume: 25(9) Issue: 9 Pages: 1056-1058
  • DOI: 10.1016/j.chembiol.2018.09.003
  • Peer Reviewed / Open Access
• [Journal Article] Colorectal cancer cells require glycogen synthase kinase-3β for sus (2018)
  • Author(s): Dewi FRP, Domoto T, Hazawa M, Kobayashi A, Douwaki T, Minamo
  • Journal Title: Oncotarget Volume: 9(17) Pages: 13337-13352
  • Peer Reviewed / Open Access
• [Journal Article] Rock-dependent phosphorylation of NUP62 regulates p63 nuclear transport and cell fate of squamous cell carcinoma. (2018)
  • Author(s): Hazawa M, Lin D, Kobayashi A, Jiang YY, Dewi FRP, Mohamed MS, Hartono H, Nakada S, Meguro-Horike M, Horike S, Koeffler H and Wong RW
  • Journal Title: EMBO Rep. Volume: 19 Issue: 1 Pages: 73-88
  • DOI: 10.15252/embr.201744523
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] High-Speed Atomic Force Microscopy Reveals Loss of Nuclear Pore Resilience as a Dying Code in Colorectal Cancer Cells. (2017)
  • Author(s): Mohamed MS, Kobayashi A, Taoka A, Watanabe-Nakayama T, Kikuchi Y, Hazawa M, Minamoto T, Fukumori Y, Kodera N, Uchihashi T, Ando T, Wong RW
  • Journal Title: ACS Nano. Volume: 11(6) Issue: 6 Pages: 5567-5578
  • DOI: 10.1021/acsnano.7b00906
  • Peer Reviewed / Open Access
• [Journal Article] ZNF750 is a lineage-specific tumor suppressor in squamous cell carcinoma. (2017)
  • Author(s): Hazawa M, Lin D, Handral H, Xu L, Chen Y, Jiang YY, Thippeswamy A, Ding LW, Meng X, Sharma A, Samuel S, Movahednia M, Wong RW, Yang H, Tong C, and Koeffler H
  • Journal Title: Oncogene Volume: 36(16) Issue: 16 Pages: 2243-2254
  • DOI: 10.1038/onc.2016.377
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] The nuclear pore complex: nanomachine as gatekeeper of molecular transport across nuclear membranes. (2018)
  • Author(s): Richard Wong
  • Organizer: The 2nd NanoLSI International Symposium - Towards Establishment of New Research Field: Nanoprobe Life Science
  • Int'l Joint Research / Invited
• [Presentation] High-speed atomic force microscopy visualization of the spatiotemporal dynamics of the nuclear nanomachine- the nuclear pore complex (2018)
  • Author(s): Richard Wong
  • Organizer: 第18回日本蛋白質科学会年会(ワークショップ)
  • Invited
• [Presentation] Molecular imaging of spatiotemporal structural dynamics of the Nuclear Pore Complex (2018)
  • Author(s): Richard Wong
  • Organizer: 新学術領域研究「動的構造生命」班会議
  • Invited
• [Presentation] Nuclear Pore Complex. (2018)
  • Author(s): Richard Wong
  • Organizer: Research Center for Advance Science & Technology Seminar, The University of Tokyo
  • Invited
• [Presentation] Disease-specific alteration of karyopherin-alpha subtypes establishes feedforward oncogenic signaling in squamous cell carcinoma. (2018)
  • Author(s): Hazawa M, Sakai K, Kobayashi A, Wong RW.
  • Organizer: THE 2018 ASCB | EMBO MEETING
  • Int'l Joint Research
• [Presentation] Nuclear transport system caused by disease-specific Karyopherin alternation (2018)
  • Author(s): Sakai K, Hazawa M, Kobayashi A, Wong RW
  • Organizer: Joint Annual Meeting of 70th JSCB and 51st JSDB
• [Presentation] Visualization spatiotemporal dynamics of RNA export factor RAE1 and RNAs complex by nanoscopic imaging. (2018)
  • Author(s): Kobayashi A, Kodera N, Morikawa K, Wong RW.
  • Organizer: The 1st WPI NanoLSI site visit
  • Int'l Joint Research
• [Presentation] The role of nucleoporin NUP58 during division (2018)
  • Author(s): Hartono H, Hazawa M, Dewi F, Kobayashi A, Mohamed MS, Wong RW.
  • Organizer: Joint Annual Meeting of 70th JSCB and 51st JSDB
• [Presentation] An oncogenic role of Tpr in Ependymoma (2018)
  • Author(s): Dewi F, Jiapaer S, Hazawa M, Sabit H, Kobayashi A, Hartono H, Nakada M, Wong RW.
  • Organizer: Joint Annual Meeting of 70th JSCB and 51st JSDB
• [Presentation] 上衣腫におけるヌクレオポリン構成分子Tprはオートファジー誘導細胞死抑制因子である (2018)
  • Author(s): 淑瑠ヘムラサビット、Jiapaer S, 玉井翔、Dewi F, 羽澤勝治、小林亜紀子、Wong RW, 中田光俊
  • Organizer: 第35回日本脳腫瘍病理学会
• [Presentation] 高速原子間力顕微鏡(HS-AFM)によるRNA輸送因子(RAE1)とRNAの動態観察 (2018)
  • Author(s): 森川亮志、古寺哲幸、小林亜紀子、Richard W. Wong
  • Organizer: 第41回日本分子生物学会年会
• [Presentation] Cisplatin-Induced Plasmid DNA Conformations Revealed by High-Speed Atomic Force Microscopy (2018)
  • Author(s): Hanbo Wang, Mahmoud Shaaban Mohamed, Kee Siang Lim, Hiroki Konnno, Takahiro Nakayama, Noriyuki Kodera, Richard W. Wong
  • Organizer: 第41回日本分子生物学会年会
• [Presentation] DNA損傷応答における核膜孔複合体の役割 (2018)
  • Author(s): 伊賀 祥紘、羽澤 勝治、Richard W. Wong
  • Organizer: 第41回日本分子生物学会年会
• [Presentation] THE SPATIOTEMPORAL DYNAMICS OF NUCLEOPORIN NUP58 DURING CELL DIVISION (2018)
  • Author(s): Hartono Hartono, Masaharu Hazawa, Firli Rahmah Primula Dewi, Akiko Kobayashi, Mahmoud Shaaban Mohamed, Lim Kee Siang, Richard W. Wong
  • Organizer: 第41回日本分子生物学会年会
• [Presentation] Spatiotemporal dynamics of the nuclear nanomachine-the nuclear pore complex. (2018)
  • Author(s): Richard Wong
  • Organizer: The 1st NanoLSI International Symposium - Towards Establishment of New Research Field: Nanoprobe Life Science
  • Int'l Joint Research / Invited
• [Presentation] Identifying nanoscopic spider cobweb features of native nuclear pores in cancer cells. (2018)
  • Author(s): Richard Wong
  • Organizer: International Symposium on Atomic Force Microscopy at Solid-Liquid Interfaces
  • Int'l Joint Research / Invited
• [Presentation] Molecular imaging of protein structure and traffic dynamics between cytoplasm and nucleus. (2018)
  • Author(s): Richard Wong
  • Organizer: The 3rd WPI NanoLSI Transdisciplinary Research Promotion Seminar
  • Invited
• [Presentation] Nuclear pore architecture and dynamics. (2018)
  • Author(s): Richard Wong
  • Organizer: 京都大学大学院工学研究科分子工学専攻生体分子機能化学セミナー
  • Invited
• [Presentation] 動的核膜孔複合体の構造と機能と病態 (2017)
  • Author(s): Richard Wong
  • Organizer: 金沢大学・北陸先端科学技術大学院大学融合科学共同専攻第２回共同ワークショップ
  • Invited
• [Presentation] Structure, dynamics and function of nuclear pore complexes (2017)
  • Author(s): Richard Wong
  • Organizer: 革新的統合バイオ研究コア ワークショップ-多様な生命現象数理モデル研究
  • Invited
• [Presentation] High-speed atomic force microscopy visualization of the nuclear pores dynamics in colon cancer cells (2017)
  • Author(s): Richard Wong
  • Organizer: 第17回日本蛋白質科学会年会
• [Presentation] 核膜孔複合体蛋白群の動的構造と機能と病態 (2017)
  • Author(s): Richard Wong
  • Organizer: 金沢大学・北陸先端科学技術大学院大学融合科学共同専攻第1回共同ワークショップ
  • Invited
• [Presentation] ROCK-dependent phosphorylation of NUP62 regulates p63 nuclear transport in squamous cell carcinoma (2017)
  • Author(s): Hazawa M, Kobayashi A, Wong RW
  • Organizer: The 2017 ASCB | EMBO Meeting
  • Int'l Joint Research
• [Presentation] Nuclear Pore Selective Barrier Dynamics as Revealed by HigSpeed Atomic Force Microscopy in Colorectal Cancer Cells (2017)
  • Author(s): Mohamed MS, Kikuchi Y, Watanabe-Nakayama T, Taoka A, Kobayashi A, Hazawa M, Kodera N, Uchihashi T, Minamoto T, Fukumori Y, Ando T, Wong RW
  • Organizer: The 2017 ASCB | EMBO Meeting
  • Int'l Joint Research
• [Presentation] Colorectal cancer cells require glycogen synthase kinase3 for sustaining mitosis via translocated promotor region (Tpr)‐dynein interaction. (2017)
  • Author(s): Dewi F, Domoto T, Hazawa M, Kobayashi A, Douwaki T, Minamoto T, Wong RW
  • Organizer: The 2017 ASCB | EMBO Meeting
  • Int'l Joint Research
• [Presentation] Nuclear Pore Complex Dynamics Revealed by High-Speed Atomic Force Microscopy in Colorectal Cancer Cells. (2017)
  • Author(s): Mohamed MS, Kikuchi Y, Watanabe-Nakayama T, Taoka A, Kobayashi A, Hazawa M, Kodera N, Uchihashi T, Minamoto T, Fukumori Y, Ando T, Wong RW
  • Organizer: International Symposium on Atomic Force Microscopy at Solid-Liquid Interfaces
  • Int'l Joint Research
• [Presentation] Loss of Nuclear Pore Selective Barrier Revealed by High-Speed Atomic Force Microscopy in Colorectal Cancer Cells (2017)
  • Author(s): Mohamed MS, Kikuchi Y, Watanabe-Nakayama T, Taoka A, Kobayashi A, Hazawa M, Kodera N, Uchihashi T, Minamoto T, Fukumori Y, Ando T, Wong RW
  • Organizer: 日本生物物理学会第55回年会
• [Presentation] 皮膚組織幹細胞におけるNup153 の役割 (2017)
  • Author(s): 塚原涼太，小林亜紀子，Wong RW，羽澤勝治
  • Organizer: 第69回細胞生物学会大会
• [Presentation] Mitotic crosstalks between translocated promoter region (Tpr) and Aurora A are key determinants of Ependymoma. (2017)
  • Author(s): Dewi F, Kobayashi A, Sabit H, Douwaki T, Dong Y, Hazawa M, Hartono, Shabierjiang J, Nakada M, Wong RW
  • Organizer: 第69回細胞生物学会大会
• [Presentation] Dynamics and Behavior of Cancer Nuclear Pore Complexes as Revealed by High-Speed Atomic Force Microscopy (2017)
  • Author(s): Mohamed MS, Kikuchi Y, Watanabe-Nakayama T, Taoka A, Kobayashi A, Hazawa M, Kodera N, Uchihashi T, Minamoto T, Fukumori Y, Ando T, Wong RW
  • Organizer: 日本生化学会北陸支部第35回大会
• [Presentation] Colorectal cancer cells require glycogen synthase kinase-3β for Sustaining mitosis via translocated promoter region(Tpr)-dynein interaction (2017)
  • Author(s): Dewi F, Domoto T, Hazawa M, Kobayashi A, Douwaki T, Minamoto T, Wong RW
  • Organizer: 日本生化学会北陸支部第35回大会
• [Book] Nuclear Pore Complexes in Genome Organization, Function and Maintenance (2018)
  • Author(s): Herausgegeben von D'Angelo, Maximiliano ( References10: Richard Wong)
  • Publisher: Springer International Publishing
  • ISBN: 9783319716121
• [Remarks] 分子ナノゲート核膜孔複合体分子が細胞の運命を決定する仕組みを解明
  • URL: https://nanolsi.kanazawa-u.ac.jp/achievements/achievements-955/
• [Remarks] 世界最速の原子間力顕微鏡を用いて大腸がん細胞核核膜孔の動きの撮影に世界で初めて成功
  • URL: https://www.kanazawa-u.ac.jp/rd/47455
• [Patent(Industrial Property Rights)] 「高速原子間力顕微鏡による細胞小器官の観察のための試料調製方法」 (2017)
  • Inventor(s): ウォング ウィン チェン リチャード 他
  • Industrial Property Rights Holder: ウォング ウィン チェン リチャード 他
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2017-223106
  • Filing Date: 2017