Collective cell migration by activation wave of growth factor signal transduction cascade

• Project/Area Number: 19H00993
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Medium-sized Section 44:Biology at cellular to organismal levels, and related fields
• Research Institution: Kyoto University
• Principal Investigator: 松田 道行 京都大学, 生命科学研究科, 教授 (10199812)
• Co-Investigator(Kenkyū-buntansha): 高橋 康史 金沢大学, ナノ生命科学研究所, 特任教授 (90624841) ; 廣田 圭昭 京都大学, 医学研究科, 助教 (20852263) ; 平島 剛志 京都大学, 医学研究科, 講師 (10620198)
• Project Period (FY): 2019-04-01 – 2024-03-31
• Project Status: Granted (Fiscal Year 2023)
• Budget Amount: ¥45,110,000 (Direct Cost: ¥34,700,000、Indirect Cost: ¥10,410,000); Fiscal Year 2023: ¥7,020,000 (Direct Cost: ¥5,400,000、Indirect Cost: ¥1,620,000); Fiscal Year 2022: ¥7,930,000 (Direct Cost: ¥6,100,000、Indirect Cost: ¥1,830,000); Fiscal Year 2021: ¥8,840,000 (Direct Cost: ¥6,800,000、Indirect Cost: ¥2,040,000); Fiscal Year 2020: ¥9,620,000 (Direct Cost: ¥7,400,000、Indirect Cost: ¥2,220,000); Fiscal Year 2019: ¥11,700,000 (Direct Cost: ¥9,000,000、Indirect Cost: ¥2,700,000)
• Keywords: 上皮細胞増殖因子 / FRETバイオセンサー / ERKマップキナーゼ / 細胞集団運動 / ERK / マップキナーゼ / 上皮細胞増殖因子受容体 / ERK MAPキナーゼ / 細胞生物学 / 顕微鏡 / バイオセンサー / 細胞運動

Outline of Research at the Start

申請者らはFRETバイオセンサーを使った研究により、上皮細胞増殖因子受容体（EGFR）を介したERKマップキナーゼの活性化が周辺の細胞に連鎖反応的に誘導されるERK活性波伝播現象を発見した。細胞集団運動時にはこのERK活性波が先導細胞から追随細胞へと伝播し、細胞運動の方向性を決定している。本研究では①細胞集団運動時のERK活性波と力学波の伝播様式の関係性の解明と数理モデル構築、②細胞膜形態変化の伝播様式のイオンコンダクタンス走査顕微鏡による解析、③細胞集団運動を司るEGFRリガンドの同定、④EGFRリガンドの拡散様式の解明、⑤生体内におけるERK活性波伝播現象の再現を行う。

Outline of Annual Research Achievements

細胞集団運動は細胞間接着を保ったまま細胞が移動する現象で、その理解は組織発生、創傷治癒、癌細胞浸潤などを理解するうえで極めて重要である。申請者らは細胞集団運動時に上皮細胞増殖因子受容体（EGFR）を介したERKマップキナーゼの活性化が先導する細胞から後方の細胞へと連鎖反応的に誘導される現象を発見している。研究開始時に、解明したい課題を5つ挙げ、目標１、３、５の研究テーマはすでに論文として発表した。目標２については、実験系の構築が期待した通りには進まなかったため中止した。残る目標4について引き続き研究を行い、全7種類のEGFRリガンドプローブおよびコントロールプローブ2種類を使って、薬剤誘導性にこれらの分子がどこまで遠く影響を及ぼしうるのかを明らかにすることができた。驚くべきことに、もっとも親和性が低いEREGが上皮細胞においてはより早くより遠くまでシグナルを伝搬しうることが明らかとなった。EREGが高親和性EGFRリガンドであるHB-EGFやTGFaよりもより効率的にERKを活性化しうるメカニズムを明らかにするために、細胞内への取り込みを共焦点顕微鏡で解析した。本研究を進めるために、EGFRリガンドの専門家であるPittsburgh大学のSorkin研究室へ大学院生を派遣し研究を進めた。さらに、この現象が生体でも起きているかを確認するために、EREGのノックアウトマウスを作成し、ここにERKバイオセンサーを発現させ、これを多光子顕微鏡で観察することで、EREGが創傷治癒過程においてERK活性伝搬に重要な役割を果たすことを明らかにした。

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

目標２は、SICMを使った研究を進める予定であったが、想定ほどの解像度が得られず、研究を断念することとなった。

Strategy for Future Research Activity

目標２以外は順調に進み、最終年度には目標４の研究を論文発表して、研究を完遂する予定である。

Research Products

• [Int'l Joint Research] University of Pittsburgh(米国)
• [Journal Article] C-type Natriuretic Peptide-induced PKA Activation Promotes Endochondral Bone Formation in Hypertrophic Chondrocytes (2022)
  • Author(s): Keisho Hirota, Tsuyoshi Hirashima, Kazuki Horikawa, Akihiro Yasoda, Michiyuki Matsuda
  • Journal Title: Endocrinology Volume: 163(3) Issue: 3 Pages: 1-12
  • DOI: 10.1210/endocr/bqac005
  • Peer Reviewed / Open Access
• [Journal Article] Functional visualization of NK cell-mediated killing of metastatic single tumor cells (2022)
  • Author(s): Ichise Hiroshi、Tsukamoto Shoko、Hirashima Tsuyoshi、Konishi Yoshinobu、Oki Choji、Tsukiji Shinya、Iwano Satoshi、Miyawaki Atsushi、Sumiyama Kenta、Terai Kenta、Matsuda Michiyuki
  • Journal Title: eLife Volume: 11
  • DOI: 10.7554/elife.76269
  • NAID: 120007191378
  • Peer Reviewed / Open Access
• [Journal Article] A feedback loop between lamellipodial extension and HGF-ERK signaling specifies leader cells during collective cell migration (2022)
  • Author(s): Hino Naoya、Matsuda Kimiya、Jikko Yuya、Maryu Gembu、Sakai Katsuya、Imamura Ryu、Tsukiji Shinya、Aoki Kazuhiro、Terai Kenta、Hirashima Tsuyoshi、Trepat Xavier、Matsuda Michiyuki
  • Journal Title: Developmental Cell Volume: 57 Issue: 19 Pages: 2290-2304
  • DOI: 10.1016/j.devcel.2022.09.003
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Visualization of Spatially-Controlled Vasospasm by Sympathetic Nerve-Mediated ROCK Activation. (2021)
  • Author(s): Ayako Imanishi, Hiroshi Ichise, Chuyun Fan, Yasuaki Nakagawa, Koichiro Kuwahara, Kenta Sumiyama, Michiyuki Matsuda, Kenta Terai.
  • Journal Title: Am J Phathol Volume: 191(1) Issue: 1 Pages: 194-203
  • DOI: 10.1016/j.ajpath.2020.09.012
  • Peer Reviewed / Open Access
• [Journal Article] Retrograde ERK activation waves drive base-to-apex multicellular flow in murine cochlear duct morphogenesis (2021)
  • Author(s): Ishii M, Tateya T, Matsuda M, Hirashima T.
  • Journal Title: eLife Volume: 10 Pages: 1-16
  • DOI: 10.7554/elife.61092
  • NAID: 120006976053
  • Peer Reviewed / Open Access
• [Journal Article] Two‐photon AMPK and ATP imaging reveals the bias between rods and cones in glycolysis utility (2021)
  • Author(s): He Jiazhou、Yamamoto Masamichi、Sumiyama Kenta、Konagaya Yumi、Terai Kenta、Matsuda Michiyuki、Sato Shinya
  • Journal Title: The FASEB Journal Volume: 35 Issue: 9
  • DOI: 10.1096/fj.202101121r
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Redundant roles of EGFR ligands in the ERK activation waves during collective cell migration (2021)
  • Author(s): Shuhao Lin, Daiki Hirayama, Gembu Maryu, Kimiya Matsuda, Naoya Hino, Eriko Deguchi, Kazuhiro Aoki, Ryo Iwamoto, Kenta Terai, Michiyuki Matsuda
  • Journal Title: Life Science Alliance Volume: 5 Issue: 1 Pages: 1-13
  • DOI: 10.26508/lsa.202101206
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] ERK-mediated mechanochemical waves direct collective cell polarization. (2020)
  • Author(s): Hino N, Rossetti L, Marin-Llaurad A, Aoki K, Trepat X, Matsuda M, Hirashima T.
  • Journal Title: Developmental Cell Volume: 53 Issue: 6 Pages: 646-660
  • DOI: 10.1016/j.devcel.2020.05.011
  • NAID: 120006876575
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Indispensable role of STIL in the regulation of cancer cell motility through the lamellipodial accumulation of ARHGEF7-PAK1 complex (2020)
  • Author(s): Hideaki Ito, Takumi Tsunoda, Miho Riku, Shingo Inaguma, Akihito Inoko, Hideki Murakami, Hiroshi Ikeda, Michiyuki Matsuda & Kenji Kasai
  • Journal Title: Oncogene Volume: 39 Issue: 9 Pages: 1931-1943
  • DOI: 10.1038/s41388-019-1115-9
  • Peer Reviewed / Open Access
• [Journal Article] Single-Cell Activation of the cAMP-Signaling Pathway in 3D Tissues with FRET-Assisted Two-Photon Activation of bPAC (2020)
  • Author(s): Kinjo Tomoaki、Watabe Tetsuya、Kobachi Kenju、Terai Kenta、Matsuda Michiyuki
  • Journal Title: ACS Chemical Biology Volume: 15 Issue: 11 Pages: 2848-2853
  • DOI: 10.1021/acschembio.0c00333
  • Peer Reviewed
• [Journal Article] Altered G1 signaling order and commitment point in cells proliferating without CDK4/6 activity (2020)
  • Author(s): Liu Chad、Konagaya Yumi、Chung Mingyu、Daigh Leighton H.、Fan Yilin、Yang Hee Won、Terai Kenta、Matsuda Michiyuki、Meyer Tobias
  • Journal Title: Nature Communications Volume: 11 Issue: 1 Pages: 5305-5305
  • DOI: 10.1038/s41467-020-18966-9
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Experimental pathology by intravital microscopy and genetically encoded fluorescent biosensors (2020)
  • Author(s): Matsuda Michiyuki、Terai Kenta
  • Journal Title: Pathology International Volume: 70 Issue: 7 Pages: 379-390
  • DOI: 10.1111/pin.12925
  • NAID: 120007052720
  • Peer Reviewed
• [Journal Article] Engineering orthogonal, plasma membrane-specific SLIPT systems for multiplexed chemical control of signaling pathways in living single cells (2020)
  • Author(s): Akinobu Nakamura, Choji Oki, Kenya Kato, Satoko Fujinuma, Gembu Maryu, Keiko Kuwata, Tatsuyuki Yoshii, Michiyuki Matsuda, Kazuhiro Aoki, Shinya Tsukiji
  • Journal Title: ACS Chemical Biology Volume: 15 Issue: 4 Pages: 1004-1015
  • DOI: 10.1021/acschembio.0c00024
  • Peer Reviewed / Open Access
• [Journal Article] Rhodopsin-mediated light-off-induced protein kinase A activation in mouse rod photoreceptor cells (2020)
  • Author(s): Sato Shinya、Yamashita Takahiro、Matsuda Michiyuki
  • Journal Title: Proceedings of the National Academy of Sciences Volume: 117 Issue: 43 Pages: 26996-27003
  • DOI: 10.1073/pnas.2009164117
  • NAID: 120007142015
  • Peer Reviewed
• [Presentation] Cell-to-cell communications as visualized by FRET biosensors (2022)
  • Author(s): Michiyuki Matsuda
  • Organizer: Research Seminar, Center for Molecular Medicine, UMC Utrecht
  • Int'l Joint Research / Invited
• [Presentation] Redundant and specific roles of epidermal growth factor receptors and their ligands in collective cell migration (2022)
  • Author(s): Michiyuki Matsuda
  • Organizer: IMPRS Distinguished Guests Seminar Series
  • Int'l Joint Research / Invited
• [Presentation] 細胞間情報伝達の可視化に基づく 病態解明 (2022)
  • Author(s): 松田道行
  • Organizer: 第29回肝細胞研究会
  • Invited
• [Presentation] FRET バイオセンサーを使った細胞間コミュニケーションの可視化 (2022)
  • Author(s): 松田 道行
  • Organizer: 学術変革領域研究（A） 新興硫黄生物学が拓く生命原理変革（硫黄生物学） 第2回 領域会議
  • Invited
• [Presentation] The Quantitative Landscape of ERK MAPK Signal Dynamics in Patient-Derived Pancreatic Cancer Organoids (2022)
  • Author(s): Shoko Tsukamoto,Akihito Machinaga,Nobuyuki Kakiuchi,Seishi Ogawa,Hiroshi Seno,Shigeki Higashiyama,Michiyuki Matsuda,Toru Hiratsuka
  • Organizer: The 7th JCA-AACR Special Joint Conference
  • Int'l Joint Research
• [Presentation] Split蛍光タンパク質を用いた多重蛍光ラベル技術 (2022)
  • Author(s): 石井衛,金城智章,寺井健太,松田道行
  • Organizer: 第74回日本細胞生物学会大会
• [Presentation] EXTRACELLULAR DYNAMICS OF EGFR LIGANDS CONTROLS THE INTERCELLULAR PROPAGATION OF ERK ACTIVATION (2022)
  • Author(s): Eriko Deguchi,Lin Shuhao,Kimiya Matsuda,Daiki Hirayama,Shinya Tsukiji,Kenta Terai,Michiyuki Matsuda
  • Organizer: FASEB The Protein Kinases and Phosphorylation Conference: Mechanisms to Therapeutics
  • Int'l Joint Research
• [Presentation] Gq-protein-coupled receptor signaling in tumor cells promotes cancer immune evasion (2020)
  • Author(s): Kenta Terai, Yoshinobu Konishi, Hiroshi Ichise, Tetsuya Watabe, Yukari Sando, Takefumi Kondo, Choji Oki, Shinya Tsukiji, Yoko Hamazaki, Yasuhiro Murakawa, Akifumi Takaori-Kondo and Michiyuki Matsuda
  • Organizer: AACR Virtual Special Conference on Tumor Heterogeneity: From Single Cells to Clinical Impact
  • Int'l Joint Research
• [Presentation] Mechano-chemical coupling via ERK signal for repetitive branching morphogenesis of lung epithelial sheet (2019)
  • Author(s): Tsuyoshi Hirashima, Takuya Yoshida, Michiyuki Matsuda
  • Organizer: EMBL Symposium: Mechanical Forces in Development
  • Int'l Joint Research
• [Presentation] Visualization of an anti-tumor immune response (2019)
  • Author(s): 寺井健太、一瀬大志、松田道行
  • Organizer: 第108回日本病理学会総会
• [Presentation] ERK activation waves mediated by intercellular mechanical forces underlie long-range guidance for collective cell migration (2019)
  • Author(s): Naoya Hino, Michiyuki Matsuda, Tsuyoshi Hirashima
  • Organizer: ASCB｜EMBO 2019 meeting
  • Int'l Joint Research
• [Remarks] 細胞の集まりからリーダーが生まれる仕組み―出る杭はより出る―
  • URL: https://www.kyoto-u.ac.jp/ja/research-news/2022-09-29-5
• [Remarks] NK細胞による転移がん細胞殺傷の可視化
  • URL: https://www.fret.lif.kyoto-u.ac.jp/rab/ichise_2022.html
• [Remarks] がん細胞が免疫から逃れるメカニズムの解明
  • URL: https://www.fret.lif.kyoto-u.ac.jp/rab/konishi_2021.html
• [Remarks] うずまき管の伸長を司る分子活性と細胞群の波を発見
  • URL: https://www.fret.lif.kyoto-u.ac.jp/rab/ishii_2020.html
• [Remarks] 血管攣縮を制御する時間・空間的な機構を解明
  • URL: https://www.fret.lif.kyoto-u.ac.jp/rab/imanishi_2020.html