Signaling basis of GPCR-acting drugs and their therapeutic mechanisms

• Project/Area Number: 17K08264
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Biological pharmacy
• Research Institution: Tohoku University
• Principal Investigator: Inoue Asuka 東北大学, 薬学研究科, 准教授 (50525813)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: GPCR / Gタンパク質共役型受容体 / バイアスリガンド / シグナル / シグナル伝達 / 薬学 / 薬理学

Outline of Final Research Achievements

G-protein-coupled receptors (GPCRs) represent the largest class of human membrane proteins. They are also regarded as important targets for drug development. GPCRs mediate signal information from external stimuli such as hormones and drugs to intracellular responses. Characterizing signal profiling of GPCRs is critical for understanding not only GPCR biology, but also GPCR-targeting drugs. In this project, we developed various methods to assess G-protein-coupling profiles of GPCRs and characterized more than 100 GPCR signaling. Through bioinformatics analysis, this experimental database enabled us to develop an algorithm to score signaling profile based on a given GPCR sequence. These achievement will contribute to development of GPCR-targeting drugs including a signal-biased ligand, which is expected to serve as a safe drug with a reduced side effect.

Academic Significance and Societal Importance of the Research Achievements

本研究を通じてGPCRがどのような分子機構を介して特定の細胞内情報（シグナル）伝達を誘導するかを解析し、分子レベルでの知見を深めた。本研究では特に、バイアスリガンドと呼ばれる特定のシグナルを選択的に作動することのできるGPCR作用薬の解析に注力し、バイアスリガンドとGPCRの結合の構造基盤を解析するとともに、バイアスリガンドと結合したGPCRがどのような分子と相互作用することでシグナル伝達が生じるかについて、その作用機序の一端を解明した。この成果を利用して、副作用のない安全な薬としてのバイアスリガンドのさらなる研究が望まれる。

Research Products

• [Int'l Joint Research] スタンフォード大学/UCSD/ハーバード大学(米国)
• [Int'l Joint Research] グラスゴー大学(英国)
• [Int'l Joint Research] ハイデルベルク大学/ボン大学(ドイツ)
• [Int'l Joint Research] モナッシュ大学(オーストラリア)
• [Int'l Joint Research] カロリンスカ研究所(スウェーデン)
• [Int'l Joint Research] Imperial College London/University of Glasgow(英国)
• [Int'l Joint Research] The University of Melbourne/The University of Queensland(オーストラリア)
• [Int'l Joint Research] University of Copenhagen(デンマーク)
• [Int'l Joint Research] NIH(米国)
• [Int'l Joint Research] グラスゴー大学(英国)
• [Int'l Joint Research] コペンハーゲン大学(デンマーク)
• [Int'l Joint Research] メルボルン大学(オーストラリア)
• [Int'l Joint Research] ボン大学(ドイツ)
• [Journal Article] The Crystal Structure of Angiotensin II Type 2 Receptor with Endogenous Peptide Hormone (2020)
  • Author(s): Asada Hidetsugu、Inoue Asuka、Ngako Kadji Francois Marie、Hirata Kunio、Shiimura Yuki、Im Dohyun、Shimamura Tatsuro、Nomura Norimichi、Iwanari Hiroko、Hamakubo Takao、Kusano-Arai Osamu、Hisano Hiromi、Uemura Tomoko、Suno Chiyo、Aoki Junken、Iwata So
  • Journal Title: Structure Volume: 28 Issue: 4 Pages: 418-425.e4
  • DOI: 10.1016/j.str.2019.12.003
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Activation of the GLP-1 receptor by a non-peptidic agonist (2020)
  • Author(s): Zhao Peishen、Liang Yi-Lynn、Belousoff Matthew J.、Deganutti Giuseppe、Fletcher Madeleine M.、Willard Francis S.、Bell Michael G.、Christe Michael E.、Sloop Kyle W.、Inoue Asuka、Truong Tin T.、Clydesdale Lachlan、Furness Sebastian G. B.、Christopoulos Arthur、Wang Ming-Wei、et al
  • Journal Title: Nature Volume: 577 Issue: 7790 Pages: 432-436
  • DOI: 10.1038/s41586-019-1902-z
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Structure of the neurotensin receptor 1 in complex with β-arrestin 1 (2020)
  • Author(s): Huang Weijiao、Masureel Matthieu、Qu Qianhui、Janetzko John、Inoue Asuka、Kato Hideaki E.、Robertson Michael J.、Nguyen Khanh C.、Glenn Jeffrey S.、Skiniotis Georgios、Kobilka Brian K.
  • Journal Title: Nature Volume: 579 Issue: 7798 Pages: 303-308
  • DOI: 10.1038/s41586-020-1953-1
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Membrane Phospholipid Analogues as Molecular Rulers to Probe the Position of the Hydrophobic Contact Point of Lysophospholipid Ligands on the Surface of G-Protein-Coupled Receptor during Membrane Approach (2020)
  • Author(s): Sayama Misa、Uwamizu Akiharu、Otani Yuko、Inoue Asuka、Aoki Junken、Sekijima Masakazu、Ohwada Tomohiko
  • Journal Title: Biochemistry Volume: 59 Issue: 11 Pages: 1173-1201
  • DOI: 10.1021/acs.biochem.0c00061
  • Peer Reviewed / Open Access
• [Journal Article] Cryo-EM structure of the human PAC1 receptor coupled to an engineered heterotrimeric G protein (2020)
  • Author(s): Kobayashi Kazuhiro、Shihoya Wataru、Nishizawa Tomohiro、Kadji Francois Marie Ngako、Aoki Junken、Inoue Asuka、Nureki Osamu
  • Journal Title: Nature Structural & Molecular Biology Volume: 27 Issue: 3 Pages: 274-280
  • DOI: 10.1038/s41594-020-0386-8
  • Peer Reviewed / Open Access
• [Journal Article] Receptor selectivity between the G proteins Gα12 and Gα13 is defined by a single leucine-to-isoleucine variation (2019)
  • Author(s): Mackenzie Amanda E.、Quon Tezz、Lin Li-Chiung、Hauser Alexander S.、Jenkins Laura、Inoue Asuka、Tobin Andrew B.、Gloriam David E.、Hudson Brian D.、Milligan Graeme
  • Journal Title: The FASEB Journal Volume: 33 Issue: 4 Pages: 5005-5017
  • DOI: 10.1096/fj.201801956r
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Illuminating the Onco-GPCRome: Novel G protein-coupled receptor-driven oncocrine networks and targets for cancer immunotherapy (2019)
  • Author(s): Wu Victoria、Yeerna Huwate、Nohata Nijiro、Chiou Joshua、Harismendy Olivier、Raimondi Francesco、Inoue Asuka、Russell Robert B.、Tamayo Pablo、Gutkind J. Silvio
  • Journal Title: Journal of Biological Chemistry Volume: 294 Issue: 29 Pages: 11062-11086
  • DOI: 10.1074/jbc.rev119.005601
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Lysolipid receptor cross-talk regulates lymphatic endothelial junctions in lymph nodes (2019)
  • Author(s): Hisano Yu、Kono Mari、Cartier Andreane、Engelbrecht Eric、Kano Kuniyuki、Kawakami Kouki、Xiong Yanbao、Piao Wenji、Galvani Sylvain、Yanagida Keisuke、Kuo Andrew、Ono Yuki、Ishida Satoru、Aoki Junken、Proia Richard L.、Bromberg Jonathan S.、Inoue Asuka、Hla Timothy
  • Journal Title: Journal of Experimental Medicine Volume: 216 Issue: 7 Pages: 1582-1598
  • DOI: 10.1084/jem.20181895
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Variable G protein determinants of GPCR coupling selectivity (2019)
  • Author(s): Okashah Najeah、Wan Qingwen、Ghosh Soumadwip、Sandhu Manbir、Inoue Asuka、Vaidehi Nagarajan、Lambert Nevin A.
  • Journal Title: Proceedings of the National Academy of Sciences of the United States of America Volume: 116 Issue: 24 Pages: 12054-12059
  • DOI: 10.1073/pnas.1905993116
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Conformational transitions of a neurotensin receptor?1?Gi1?complex (2019)
  • Author(s): Kato Hideaki E.、Zhang Yan、Hu Hongli、Suomivuori Carl-Mikael、Kadji Francois Marie Ngako、Aoki Junken、Krishna Kumar Kaavya、Fonseca Rasmus、Hilger Daniel、Huang Weijiao、Latorraca Naomi R.、Inoue Asuka、Dror Ron O.、Kobilka Brian K.、Skiniotis Georgios
  • Journal Title: Nature Volume: 572 Issue: 7767 Pages: 80-85
  • DOI: 10.1038/s41586-019-1337-6
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] WNT-3A-induced β-catenin signaling does not require signaling through heterotrimeric G proteins (2019)
  • Author(s): Bowin Carl-Fredrik、Inoue Asuka、Schulte Gunnar
  • Journal Title: Journal of Biological Chemistry Volume: 294 Issue: 31 Pages: 11677-11684
  • DOI: 10.1074/jbc.ac119.009412
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Crystal structure of human endothelin ETB receptor in complex with peptide inverse agonist IRL2500 (2019)
  • Author(s): Nagiri Chisae、Shihoya Wataru、Inoue Asuka、Kadji Francois Marie Ngako、Aoki Junken、Nureki Osamu
  • Journal Title: Communications Biology Volume: 2 Issue: 1 Pages: 1-11
  • DOI: 10.1038/s42003-019-0482-7
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] PRECOG: PREdicting COupling probabilities of G-protein coupled receptors (2019)
  • Author(s): Singh Gurdeep、Inoue Asuka、Gutkind J Silvio、Russell Robert B、Raimondi Francesco
  • Journal Title: Nucleic Acids Research Volume: 47 Issue: W1 Pages: W395-W401
  • DOI: 10.1093/nar/gkz392
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Illuminating G-Protein-Coupling Selectivity of GPCRs (2019)
  • Author(s): Inoue Asuka、Raimondi Francesco、Kadji Francois Marie Ngako、Singh Gurdeep、Kishi Takayuki、Uwamizu Akiharu、Ono Yuki、Shinjo Yuji、Ishida Satoru、Arang Nadia、Kawakami Kouki、Gutkind J. Silvio、Aoki Junken、Russell Robert B.
  • Journal Title: Cell Volume: 177 Issue: 7 Pages: 1933-1947.e25
  • DOI: 10.1016/j.cell.2019.04.044
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Rare, functional, somatic variants in gene families linked to cancer genes: GPCR signaling as a paradigm (2019)
  • Author(s): Raimondi Francesco、Inoue Asuka、Kadji Francois M. N.、Shuai Ni、Gonzalez Juan-Carlos、Singh Gurdeep、de la Vega Alicia Alonso、Sotillo Rocio、Fischer Bernd、Aoki Junken、Gutkind J. Silvio、Russell Robert B.
  • Journal Title: Oncogene Volume: 38 Issue: 38 Pages: 6491-6506
  • DOI: 10.1038/s41388-019-0895-2
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Structures of the 5-HT2A receptor in complex with the antipsychotics risperidone and zotepine (2019)
  • Author(s): Kimura Kanako Terakado、Asada Hidetsugu、Inoue Asuka、Kadji Francois Marie Ngako、Im Dohyun、Mori Chihiro、Arakawa Takatoshi、Hirata Kunio、Nomura Yayoi、Nomura Norimichi、Aoki Junken、Iwata So、Shimamura Tatsuro
  • Journal Title: Nature Structural & Molecular Biology Volume: 26 Issue: 2 Pages: 121-128
  • DOI: 10.1038/s41594-018-0180-z
  • Peer Reviewed
• [Journal Article] Multi-Component Mechanism of H2 Relaxin Binding to RXFP1 through NanoBRET Kinetic Analysis (2019)
  • Author(s): Hoare Bradley L.、Bruell Shoni、Sethi Ashish、Gooley Paul R.、Lew Michael J.、Hossain Mohammed A.、Inoue Asuka、Scott Daniel J.、Bathgate Ross A.D.
  • Journal Title: iScience Volume: 11 Pages: 93-113
  • DOI: 10.1016/j.isci.2018.12.004
• [Journal Article] Genome Editing Provides New Insights into Receptor Controlled Signalling Pathways. (2018)
  • Author(s): Milligan G, Inoue A.
  • Journal Title: Trends Pharmacol Sci. Volume: in press Issue: 5 Pages: 481-493
  • DOI: 10.1016/j.tips.2018.02.005
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Human GIP(3-30)NH 2 inhibits G protein-dependent as well as G protein-independent signaling and is selective for the GIP receptor with high-affinity binding to primate but not rodent GIP receptors (2018)
  • Author(s): Gabe Maria Buur Nordskov、Sparre-Ulrich Alexander Hovard、Pedersen Mie Fabricius、Gasbjerg L?rke Smidt、Inoue Asuka、Br?uner-Osborne Hans、Hartmann Bolette、Rosenkilde Mette Marie
  • Journal Title: Biochemical Pharmacology Volume: 150 Pages: 97-107
  • DOI: 10.1016/j.bcp.2018.01.040
• [Journal Article] BRET-based assay to monitor EGFR transactivation by the AT1R reveals Gq/11 protein-independent activation and AT1R-EGFR complexes (2018)
  • Author(s): O'Brien Shannon L.、Johnstone Elizabeth K.M.、Devost Dominic、Conroy Jacinta、Reichelt Melissa E.、Purdue Brooke W.、Ayoub Mohammed A.、Kawai Tatsuo、Inoue Asuka、Eguchi Satoru、H?bert Terence E.、Pfleger Kevin D.G.、Thomas Walter G.
  • Journal Title: Biochemical Pharmacology Volume: 158 Pages: 232-242
  • DOI: 10.1016/j.bcp.2018.10.017
• [Journal Article] Crystal structures of human ETB receptor provide mechanistic insight into receptor activation and partial activation (2018)
  • Author(s): Shihoya Wataru、Izume Tamaki、Inoue Asuka、Yamashita Keitaro、Kadji Francois Marie Ngako、Hirata Kunio、Aoki Junken、Nishizawa Tomohiro、Nureki Osamu
  • Journal Title: Nature Communications Volume: 9 Issue: 1 Pages: 1-11
  • DOI: 10.1038/s41467-018-07094-0
  • Peer Reviewed / Open Access
• [Journal Article] Real-time examination of cAMP activity at relaxin family peptide receptors using a BRET-based biosensor (2018)
  • Author(s): Valkovic Adam L.、Leckey Miranda B.、Whitehead Alice R.、Hossain Mohammed A.、Inoue Asuka、Kocan Martina、Bathgate Ross A. D.
  • Journal Title: Pharmacology Research & Perspectives Volume: 6 Issue: 5
  • DOI: 10.1002/prp2.432
  • NAID: 120006873133
• [Journal Article] Targeting GLP-1 receptor trafficking to improve agonist efficacy (2018)
  • Author(s): Jones Ben、Buenaventura Teresa、Kanda Nisha、Chabosseau Pauline、Owen Bryn M.、Scott Rebecca、Goldin Robert、Angkathunyakul Napat、Corr?a Jr Ivan R.、Bosco Domenico、Johnson Paul R.、Piemonti Lorenzo、Marchetti Piero、Shapiro A. M. James、Cochran Blake J.、Hanyaloglu Aylin C.、Inoue Asuka、et al.
  • Journal Title: Nature Communications Volume: 9 Issue: 1 Pages: 1-17
  • DOI: 10.1038/s41467-018-03941-2
• [Journal Article] On the G protein-coupling selectivity of the native A2B adenosine receptor. (2018)
  • Author(s): Gao ZG, Inoue A, Jacobson KA
  • Journal Title: Biochem Pharmacol. Volume: 39 Pages: 481-493
  • DOI: 10.1016/j.bcp.2017.12.003
  • Peer Reviewed / Int'l Joint Research
• [Presentation] GPCR配列にコードされるGタンパク質シグナル選択性の解明 (2019)
  • Author(s): 井上 飛鳥
  • Organizer: 第92回日本生化学会大会
• [Presentation] Decoding G-protein Coupling Selectivity with a Large Functional Dataset (2019)
  • Author(s): Asuka Inoue
  • Organizer: GPCR Workshop 2019
  • Int'l Joint Research / Invited
• [Presentation] A crystal structure of the lysophospholipid receptor LPA6and insights into lipid ligand entry and a hereditary hair disease (2018)
  • Author(s): Asuka Inoue
  • Organizer: International GPCR Symposium
  • Int'l Joint Research / Invited
• [Presentation] スプリットルシフェラーゼを用いた三量体Gタンパク質活性化の検出法の確立とGPCR研究への応用 (2018)
  • Author(s): 井上飛鳥、Francois Marie Ngako Kadji、青木淳賢
  • Organizer: 第91回日本生化学会大会
• [Presentation] NanoBiT-based G protein sensors (2017)
  • Author(s): Asuka Inoue, FMN Kadji, Junken Aoki
  • Organizer: GPCR Workshop 2017
  • Int'l Joint Research
• [Presentation] 三量体Gタンパク質の活性化センサーの開発 (2017)
  • Author(s): 井上 飛鳥、Francois Marie Marie Kadji、川上 耕季、青木 淳賢
  • Organizer: 2017年度生命科学系学会合同年次大会（ConBio2017）
• [Remarks] 研究室ホームページ
  • URL: http://www.pharm.tohoku.ac.jp/~seika/H28/index.html
• [Remarks] 東北大学大学院薬学研究科分子細胞生化学分野
  • URL: http://www.pharm.tohoku.ac.jp/~seika/H28/index.html