Functional regulation of ubiquitin chains using chemo-technologies

• Project Area: New frontier for ubiquitin biology driven by chemo-technologies
• Project/Area Number: 18H05499
• 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: Kyoto University
• Principal Investigator: 岩井 一宏 京都大学, 医学研究科, 教授 (60252459)
• Co-Investigator(Kenkyū-buntansha): 二木 史朗 京都大学, 化学研究所, 教授 (50199402)
• Project Period (FY): 2018-06-29 – 2023-03-31
• Project Status: Granted (Fiscal Year 2022)
• Budget Amount: ¥136,760,000 (Direct Cost: ¥105,200,000、Indirect Cost: ¥31,560,000); Fiscal Year 2022: ¥28,860,000 (Direct Cost: ¥22,200,000、Indirect Cost: ¥6,660,000); Fiscal Year 2021: ¥28,860,000 (Direct Cost: ¥22,200,000、Indirect Cost: ¥6,660,000); Fiscal Year 2020: ¥28,860,000 (Direct Cost: ¥22,200,000、Indirect Cost: ¥6,660,000); Fiscal Year 2019: ¥28,860,000 (Direct Cost: ¥22,200,000、Indirect Cost: ¥6,660,000); Fiscal Year 2018: ¥21,320,000 (Direct Cost: ¥16,400,000、Indirect Cost: ¥4,920,000)
• Keywords: ユビキチン / ステープルペプチド / ユビキチンシグナル / 炎症シグナル / 直鎖状ユビキチン鎖

Outline of Annual Research Achievements

本研究ではケモテクノロジーを用いて5年間の研究期間中にユビキチン修飾の多様な機能を解析することを目指す。4年目である本年度は以下の3項目から研究を推進した。
１．ケモテクノロジーを用いたLUBACリガーゼの制御とその応用展開：研究代表者らはLUBACによる直鎖状ユビキチン鎖生成はB細胞リンパ腫発症に寄与することを示していたが、他の計画研究の代表である吉田と開発したLUBAC阻害剤を用いて、LUBACがBリンパ腫の増殖を抑制することを個体レベルで示した（Jo et al. Blood 2020）。さらに、LUBAC複合体の結晶構造を踏まえた新規LUBAC抑制剤の開発も進めている。これまでの解析でリード化合物が取得できなかったので、AI創薬に着手した。
2．ユビキチン修飾の新機能のケモテクノロジー解析：M1鎖とUBDの結合阻害の試験管内スクリーニング系でM1鎖とHOIL-1L NZFの結合を阻害する化合物を同定した。さらに基礎的検討で同化合物が細胞内でもM1鎖とHOIL-1L NZFの結合を阻害する可能性を見出している。また、HOIL-1L、SHARPINのNZFと直鎖状ユビキチン鎖との結合阻害によって、刺激依存的に細胞を死に導くことができることを示す予備的知見を得た。
3． LUBACはHOIP、HOIL-1L、SHARPINの3サブユニットから構成され、活性中心であるHOIPだけでなく、HOIL-1Lもユビキチンリガーゼ活性を持つ。本年度、研究代表者らはHOIL-1Lリガーゼ活性を欠失がLUBACのM1鎖形成能を亢進させて細胞内感染菌の感染を抑制することを見出した。そこで、HOIL-1Lユビキチンリガーゼ活性阻害剤のスクリーニング系を構築して、吉田とともに化合物の一次スクリーニングを施行し、LUBAC賦活剤の生成に挑んでいる。

Current Status of Research Progress

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

Reason

本領域の吉田とともに開発してきた個体に投与可能なLUBAC阻害剤がB細胞リンパ腫の治療薬となることのPOCが取得できたので、本領域でより副作用の少ない阻害剤開発を推進している。

Strategy for Future Research Activity

ケミカルツールを用いたユビキチン系の解析をより一層推進する。

Research Products

• [Int'l Joint Research] North Western University(米国)
• [Int'l Joint Research] Washington University/University of Illinois at Chicago(米国)
• [Int'l Joint Research] National Center of Biomedical Analysis(中国)
• [Journal Article] Glycogen synthase downregulation rescues the amylopectinosis of murine RBCK1 deficiency (2022)
  • Author(s): Nitschke S, Sullivan MA, Mitra S, Marchioni CR, Lee JPY, Smith BH, Ahonen S, Wu J, Chown EE, Wang P, Petkovic S, Zhao X, DiGiovanni LF, Perri AM, Israelian L, Grossman TR, Kordasiewicz H, Vilaplana F, Iwai K, Nitschke F, Minassian BA
  • Journal Title: Brain Volume: In press Issue: 7 Pages: 2361-2377
  • DOI: 10.1093/brain/awac017
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] ABIN1 is a signal‐induced autophagy receptor that attenuates NF‐κB activation by recognizing linear ubiquitin chains (2022)
  • Author(s): Shinkawa Y, Imami K, Fuseya Y, Sasaki K, Ohmura K, Ishihama Y, Morinobu A, Iwai K.
  • Journal Title: FEBS Letters Volume: In press Issue: 9 Pages: 1147-1164
  • DOI: 10.1002/1873-3468.14323
  • Peer Reviewed
• [Journal Article] LUBAC-mediated linear ubiquitination: a crucial regulator of immune signaling (2021)
  • Author(s): IWAI Kazuhiro
  • Journal Title: Proceedings of the Japan Academy, Series B Volume: 97 Issue: 3 Pages: 120-133
  • DOI: 10.2183/pjab.97.007
  • NAID: 130007996827
  • ISSN: 0386-2208, 1349-2896
  • Year and Date: 2021-03-11
  • Peer Reviewed
• [Journal Article] Loss of peptide: N -glycanase causes proteasome dysfunction mediated by a sugar-recognizing ubiquitin ligase (2021)
  • Author(s): Yoshida Yukiko、Asahina Makoto、Murakami Arisa、Kawawaki Junko、Yoshida Meari、Fujinawa Reiko、Iwai Kazuhiro、Tozawa Ryuichi、Matsuda Noriyuki、Tanaka Keiji、Suzuki Tadashi
  • Journal Title: Proceedings of the National Academy of Sciences Volume: 118 Issue: 27
  • DOI: 10.1073/pnas.2102902118
  • Peer Reviewed / Open Access
• [Journal Article] Expression, solubility monitoring, and purification of the co-folded LUBAC LTM domain by structure-guided tandem folding in autoinducing cultures (2021)
  • Author(s): Walinda Erik、Morimoto Daichi、Sorada Tomoki、Iwai Kazuhiro、Sugase Kenji
  • Journal Title: Protein Expression and Purification Volume: 187 Pages: 105953-105953
  • DOI: 10.1016/j.pep.2021.105953
  • Peer Reviewed
• [Journal Article] Liquid Droplet Formation and Facile Cytosolic Translocation of IgG in the Presence of Attenuated Cationic Amphiphilic Lytic Peptides (2021)
  • Author(s): Iwata T, Hirose H, Sakamoto K, Hirai Y, Arafiles JVV, Akishiba M, Imanishi M, Futaki S
  • Journal Title: Angewandte Chemie International Edition Volume: 60 Issue: 36 Pages: 19804-19812
  • DOI: 10.1002/anie.202105527
  • Peer Reviewed
• [Journal Article] Structural Dynamic Heterogeneity of Polyubiquitin Subunits Affects Phosphorylation Susceptibility (2021)
  • Author(s): Morimoto Daichi、Walinda Erik、Takashima Shingo、Nishizawa Mayu、Iwai Kazuhiro、Shirakawa Masahiro、Sugase Kenji
  • Journal Title: Biochemistry Volume: 60 Issue: 8 Pages: 573-583
  • DOI: 10.1021/acs.biochem.0c00619
  • Peer Reviewed
• [Journal Article] Peptide-assisted Intracellular Delivery of Biomacromolecules (2020)
  • Author(s): Futaki Shiroh、Arafiles Jan Vincent V.、Hirose Hisaaki
  • Journal Title: Chemistry Letters Volume: 49 Issue: 9 Pages: 1088-1094
  • DOI: 10.1246/cl.200392
  • NAID: 130007897056
  • ISSN: 0366-7022, 1348-0715
  • Year and Date: 2020-09-05
  • Peer Reviewed / Open Access
• [Journal Article] The HOIL-1L ligase modulates immune signalling and cell death via monoubiquitination of LUBAC (2020)
  • Author(s): Fuseya Yasuhiro、Fujita Hiroaki、Kim Minsoo、Ohtake Fumiaki、Nishide Akira、Sasaki Katsuhiro、Saeki Yasushi、Tanaka Keiji、Takahashi Ryosuke、Iwai Kazuhiro
  • Journal Title: Nature Cell Biology Volume: － Issue: 6 Pages: 663-673
  • DOI: 10.1038/s41556-020-0517-9
  • Peer Reviewed / Open Access
• [Journal Article] LUBAC accelerates B-cell lymphomagenesis by conferring resistance to genotoxic stress on B cells (2020)
  • Author(s): Jo Tomoyasu、Nishikori Momoko、Kogure Yasunori、Arima Hiroshi、Sasaki Katsuhiro、Sasaki Yoshiteru、Nakagawa Tomoko、Iwai Fumie、Momose Shuji、Shiraishi Aki、Kiyonari Hiroshi、Kagaya Noritaka、Onuki Tetsuo、Shin-ya Kazuo、Yoshida Minoru、Kataoka Keisuke、Ogawa Seishi、Iwai Kazuhiro、Takaori-Kondo Akifumi
  • Journal Title: Blood Volume: 136 Issue: 6 Pages: 684-697
  • DOI: 10.1182/blood.2019002654
  • Peer Reviewed / Open Access
• [Journal Article] Discovery of linear ubiquitination, a crucial regulator for immune signaling and cell death (2020)
  • Author(s): Iwai Kazuhiro
  • Journal Title: The FEBS Journal Volume: 288 Issue: 4 Pages: 1060-1069
  • DOI: 10.1111/febs.15471
  • Peer Reviewed
• [Journal Article] Linear ubiquitin assembly complex regulates lung epithelial-driven responses during influenza infection (2020)
  • Author(s): Brazee Patricia L.、Morales-Nebreda Luisa、Magnani Natalia D.、Garcia Joe G.N.、Misharin Alexander V.、Ridge Karen M.、Budinger G.R. Scott、Iwai Kazuhiro、Dada Laura A.、Sznajder Jacob I.
  • Journal Title: Journal of Clinical Investigation Volume: 130 Issue: 3 Pages: 1301-1314
  • DOI: 10.1172/jci128368
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Stimulating Macropinocytosis for Intracellular Nucleic Acid and Protein Delivery: A Combined Strategy with Membrane-Lytic Peptides To Facilitate Endosomal Escape (2020)
  • Author(s): Arafiles Jan Vincent V.、Hirose Hisaaki、Akishiba Misao、Tsuji Shogo、Imanishi Miki、Futaki Shiroh
  • Journal Title: Bioconjugate Chemistry Volume: 31 Issue: 3 Pages: 547-553
  • DOI: 10.1021/acs.bioconjchem.0c00064
  • Peer Reviewed
• [Journal Article] Modulation of autoimmune pathogenesis by T cell-triggered inflammatory cell death. (2019)
  • Author(s): Sasaki, K., Himeno, A., Nakagawa, T., Sasaki, Y., Kiyonari, H. and Iwai, K.
  • Journal Title: Nature Communications Volume: 10 Issue: 1 Pages: 3878-3878
  • DOI: 10.1038/s41467-019-11858-7
  • NAID: 120006716601
  • Peer Reviewed / Open Access
• [Journal Article] LUBAC controls chromosome alignment by targeting CENP-E to attached kinetochores (2019)
  • Author(s): Wu Min、Chang Yan、Hu Huaibin、Mu Rui、Zhang Yucheng、Qin Xuanhe、Duan Xiaotao、Li Weihua、Tu Haiqing、Zhang Weina、Wang Guang、Han Qiuying、Li Ailing、Zhou Tao、Iwai Kazuhiro、Zhang Xuemin、Li Huiyan
  • Journal Title: Nat. Commun. Volume: 10 Issue: 1 Pages: 273-273
  • DOI: 10.1038/s41467-018-08043-7
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] HOIL1 Is Essential for the Induction of Type I and III Interferons by MDA5 and Regulates Persistent Murine Norovirus Infection (2018)
  • Author(s): MacDuff Donna A.、Baldridge Megan T.、Qaqish Arwa M.、Nice Timothy J.、Darbandi Azad D.、 Hartley Victoria L.、Peterson Stefan T.、Miner Jonathan J.、Iwai Kazuhiro、Virgin Herbert W.
  • Journal Title: Journal of Virology Volume: 92 Issue: 23
  • DOI: 10.1128/jvi.01368-18
  • Peer Reviewed / Int'l Joint Research
• [Presentation] Linear ubiquitination : a new regulator of chronic inflammation and oncogenesis (2021)
  • Author(s): Iwai, K.
  • Organizer: 第44日本基礎老化学会大会 International Symposium “Mechanism of aging”
  • Int'l Joint Research / Invited
• [Presentation] 直鎖状ユビキチン鎖の発見とその機能 (2021)
  • Author(s): 岩井 一宏
  • Organizer: 第94回日本生化学会大会シンポジウム 「プロテオスタシスの理解と革新的医療の創出」
• [Presentation] 炎症応答を制御するユニークなユビキチン修飾系と炎症性疾患への寄与 (2021)
  • Author(s): 岩井 一宏
  • Organizer: 第30回関西ライフサイエンス リーディングサイエンティストセミナー
  • Invited
• [Presentation] Attenuated cationic lytic peptides for intracellular delivery (2021)
  • Author(s): Futaki, S.
  • Organizer: The 2021 International Chemical Congress of Pacific Basin Societies (Pacifichem 2021)
  • Int'l Joint Research / Invited
• [Presentation] LUBACの2つのユビキチンリガーゼ活性による免疫シグナル制御 (2020)
  • Author(s): 岩井 一宏
  • Organizer: 第45回医用マススペクトル学会年会 シンポジウム
  • Invited
• [Presentation] 直鎖状ユビキチン鎖：免疫疾患、がんに寄与する新規シグナル伝達系 (2020)
  • Author(s): 岩井 一宏
  • Organizer: 第70回日本薬学会関西支部大会 特別講演
  • Invited
• [Presentation] From Bench to Bedsides:ユビキチン研究の進展～COVID-19創薬の可能性も含めて～ (2020)
  • Author(s): 岩井 一宏
  • Organizer: 第68回日本職業災害医学会学術大会 特別講演
  • Invited
• [Presentation] Attenuated cationic lytic peptides for intracellular delivery (2020)
  • Author(s): Futaki, S
  • Organizer: Controlled Release Society 2020 Annual Meeting & Exposition
  • Invited
• [Presentation] がん、免疫疾患を惹起するユビキチン・シグナリング系：ケミカルバイオロジーを用いた介入の可能性 (2019)
  • Author(s): 岩井 一宏
  • Organizer: 日本生化学会 近畿地方会
  • Invited
• [Presentation] ユビキチン研究の現況と創薬への応用 (2019)
  • Author(s): 岩井 一宏
  • Organizer: 第21回日本応用薬理シンポジウム
  • Invited
• [Presentation] Redesigning cationic lytic peptides to promote the delivery of biomacromolecules into cell interiors (2019)
  • Author(s): Futaki, S.
  • Organizer: American Chemical Society Fall 2019 National Meeting & Exposition
  • Int'l Joint Research / Invited
• [Remarks] 京都大学・大学院医学研究科・細胞機能制御学
  • URL: http://www.mcp-kyoto-u.jp