Construction of DUB MAP for innate immune and inflammatory responses

• Project/Area Number: 19K22541
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 49:Pathology, infection/immunology, and related fields
• Research Institution: Osaka City University
• Principal Investigator: Tokunaga Fuminori 大阪市立大学, 大学院医学研究科, 教授 (00212069)
• Project Period (FY): 2019-06-28 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000); Fiscal Year 2020: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000); Fiscal Year 2019: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
• Keywords: 脱ユビキチン化酵素 / タンパク質 / 細胞 / 炎症 / 自然免疫 / オートファジー / 酵素 / シグナル伝達 / ユビキチン / 細胞死

Outline of Research at the Start

申請者らが独創的に構築したヒト脱ユビキチン化酵素(DUB) cDNAセットを用いて、gain-of-function法にて炎症や自然免疫に重要なシグナル伝達や選択的オートファジー制御に関わるDUBの定量的・網羅的探索に挑戦するとともに、酵素機能の生化学的解析、細胞・個体レベルでの生理機能解析、DUB阻害剤の同定を目指し、炎症・自然免疫を制御するDUBの相対位置付け(DUBマップ)を明らかにする。また、シグナル伝達経路・選択的オートファジーの研究試薬や癌・炎症性疾患に対する創薬シーズの開発を目指す。本研究から、基礎医学研究のみならず、癌や炎症性疾患に対する新規創薬標的の同定が期待される。

Outline of Final Research Achievements

Protein ubiquitination was identified as a marker that binds to proteins in cells and selectively leads to proteasomal degradation. The discoverers of protein ubiquitination is awarded to the Nobel Prize in Chemistry in 2004, due to the importance of cellular functions. Subsequent studies have shown that it is involved in various cell functions, such as inflammation and immune signals, selective autophagy, DNA repair, and proteolysis. There are about 100 types of deubiquitinating enzymes (DUBs) in humans that cleave ubiquitin chains and control various cell functions. In this study, we prepared 88 types of DUBs comprehensively investigated the characteristics of DUBs involved in inflammation / immune signals and selective autophagy, and proceeded with the DUB inhibitors.

Academic Significance and Societal Importance of the Research Achievements

ユビキチン修飾系の制御破綻は癌や神経変性疾患など各種疾患発症に関わるため、近年、脱ユビキチン化酵素(DUB)が創薬標的として高く注目されている。我々は独創的にDUBのライブラリーを調整し、各種細胞機能を制御するDUBの抽出を進め、特に炎症応答に関わる新規DUBを見出すことができた。また、細胞内侵入細菌が引き起こす選択的オートファジーとの関連、及び直鎖状ユビキチンを特異的に分解するDUBに対する阻害剤探索を行なった。これらの研究から新規細胞制御性DUBの発見と創薬シーズ展開が期待できる。

Research Products

• [Journal Article] Th2 cell-derived histamine is involved in nasal Th2 infiltration in mice (2021)
  • Author(s): Iwasaki N, Terawaki S, Shimizu K, Oikawa D, Sakamoto H, Sunami K, Tokunaga F.
  • Journal Title: Inflammation Research Volume: In press Issue: 5 Pages: 539-541
  • DOI: 10.1007/s00011-021-01458-x
  • Peer Reviewed / Open Access
• [Journal Article] Th2 cells and macrophages cooperatively induce allergic inflammation through histamine signaling (2021)
  • Author(s): Iwasaki N, Terawaki S, Shimizu K, Oikawa D, Sakamoto H, Sunami K, Tokunaga F.
  • Journal Title: Plos One Volume: - Issue: 3 Pages: e0248158-e0248158
  • DOI: 10.1371/journal.pone.0248158
  • Peer Reviewed / Open Access
• [Journal Article] Crosstalk Between NDP52 and LUBAC in Innate Immune Responses, Cell Death, and Xenophagy. (2021)
  • Author(s): Miyashita H, Oikawa D, Terawaki S, Kabata D, Shintani A, Tokunaga F.
  • Journal Title: Frontiers in Immunology Volume: 12 Pages: 635475-635475
  • DOI: 10.3389/fimmu.2021.635475
  • Peer Reviewed / Open Access
• [Journal Article] 6His-tagged Recombinant Human Cytoglobin Deactivates Hepatic Stellate Cells and Inhibits Liver Fibrosis by Scavenging Reactive Oxygen Species (2021)
  • Author(s): Ninh Quoc Dat, Le Thi Thanh Thuy, Vu Ngoc Hieu, Fuminori Tokunaga, Massimo Pinzani, and Norifumi Kawada
  • Journal Title: Hepatology Volume: 0-0 Issue: 6 Pages: 0-0
  • DOI: 10.1002/hep.31752
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] MIND bomb 2 prevents RIPK1 kinase activity-dependent and -independent apoptosis through ubiquitylation of cFLIPL (2021)
  • Author(s): Nakabayashi Osamu、Takahashi Hirotaka、Moriwaki Kenta、Komazawa-Sakon Sachiko、Ohtake Fumiaki、Murai Shin、Tsuchiya Yuichi、Koyahara Yuki、Saeki Yasushi、Yoshida Yukiko、Yamazaki Soh、Tokunaga Fuminori、Sawasaki Tatsuya、Nakano Hiroyasu
  • Journal Title: Communications Biology Volume: 4 Issue: 1 Pages: 80-80
  • DOI: 10.1038/s42003-020-01603-y
  • Peer Reviewed / Open Access
• [Journal Article] The synchronized gene expression of retrotransposons and type I interferon in dermatomyositis. (2021)
  • Author(s): Kuriyama Y, Shimizu A, Kanai S, Oikawa D, Tokunaga F, Tsukagoshi H, Ishikawa O.
  • Journal Title: Journal of American Academy of Dermatology Volume: 84 Issue: 4 Pages: 1103-1105
  • DOI: 10.1016/j.jaad.2020.05.051
  • Peer Reviewed / Open Access
• [Journal Article] Involvement of linear ubiquitination in neurodegenerative diseases and development of LUBAC inhibitors (2020)
  • Author(s): 及川 大輔、伊東 秀文、徳永 文稔
  • Journal Title: 生化学 Volume: 92 Issue: 1 Pages: 28-34
  • DOI: 10.14952/SEIKAGAKU.2020.920028
  • ISSN: 0037-1017
  • Year and Date: 2020-02-25
  • Peer Reviewed
• [Journal Article] Development and application of a protein array of human deubiquitinating enzyme (2020)
  • Author(s): 高橋宏隆、山中聡士、徳永文稔、澤崎達也
  • Journal Title: 生化学 Volume: 92 Issue: 1 Pages: 64-74
  • DOI: 10.14952/SEIKAGAKU.2020.920064
  • ISSN: 0037-1017
  • Year and Date: 2020-02-25
  • Peer Reviewed
• [Journal Article] Cellular and Mathematical Analyses of LUBAC Involvement in T Cell Receptor-Mediated NF-κB Activation Pathway (2020)
  • Author(s): D.Oikawa, N.Hatanaka, T.Suzuki, F.Tokunaga
  • Journal Title: Frontiers in Immunology Volume: 11 Pages: 601926-601926
  • DOI: 10.3389/fimmu.2020.601926
  • Peer Reviewed / Open Access
• [Journal Article] A Human DUB Protein Array for Clarification of Linkage Specificity of Polyubiquitin Chain and Application to Evaluation of Its Inhibitors (2020)
  • Author(s): Takahashi Hirotaka、Yamanaka Satoshi、Kuwada Shohei、Higaki Kana、Kido Kohki、Sato Yusuke、Fukai Shuya、Tokunaga Fuminori、Sawasaki Tatsuya
  • Journal Title: Biomedicines Volume: 8 Issue: 6 Pages: 152-152
  • DOI: 10.3390/biomedicines8060152
  • Peer Reviewed / Open Access
• [Journal Article] Linear ubiquitin code: Its writer, erasers, decoders, inhibitors, and implications in disorders. (2020)
  • Author(s): Oikawa D, Sato Y, Ito H, and Tokunaga F.
  • Journal Title: International Journal of Molecular Sciences Volume: 21 Issue: 9 Pages: 3381-3381
  • DOI: 10.3390/ijms21093381
  • Peer Reviewed / Open Access
• [Journal Article] Molecular bases for HOIPINs-mediated inhibition of LUBAC and innate immune responses. (2020)
  • Author(s): Oikawa D, Sato Y, Ohtake F, Komakura K, Hanada K, Sugawara K, Terawaki S, Mizukami Y, Phuong HT, Iio K, Obika S, Fukushi M, Irie T, Tsuruta D, Sakamoto S, Tanaka K, Saeki Y, Fukai S, Tokunaga F.
  • Journal Title: Commun. Biol. Volume: 3 Issue: 1 Pages: 163-163
  • DOI: 10.1038/s42003-020-0882-8
  • Peer Reviewed / Open Access
• [Journal Article] Subquinocin, a small molecule inhibitor of CYLD and USP-family deubiquitinating enzymes, promotes NF-κB signaling (2020)
  • Author(s): Yamanaka Satoshi、Sato Yusuke、Oikawa Daisuke、Goto Eiji、Fukai Shuya、Tokunaga Fuminori、Takahashi Hirotaka、Sawasaki Tatsuya
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 524 Issue: 1 Pages: 1-7
  • DOI: 10.1016/j.bbrc.2019.12.049
  • Peer Reviewed / Open Access
• [Journal Article] 創薬を見据えた直鎖状ユビキチン鎖生成酵素(LUBAC)阻害剤開発 (2020)
  • Author(s): 徳永 文稔
  • Journal Title: Farumashia Volume: 56 Issue: 1 Pages: 26-30
  • DOI: 10.14894/faruawpsj.56.1_26
  • NAID: 130007779035
  • ISSN: 0014-8601, 2189-7026
  • Peer Reviewed
• [Journal Article] Linear Polyubiquitin Chain Modification of TDP-43-Positive Neuronal Cytoplasmic Inclusions in Amyotrophic Lateral Sclerosis (2019)
  • Author(s): Nakayama Yoshiaki、Tsuji Kazumi、Ayaki Takashi、Mori Megumi、Tokunaga Fuminori、Ito Hidefumi
  • Journal Title: Journal of Neuropathology & Experimental Neurology Volume: 79 Issue: 3 Pages: 256-265
  • DOI: 10.1093/jnen/nlz135
  • Peer Reviewed
• [Journal Article] The E3 ubiquitin ligase MIB2 enhances inflammation by degrading the deubiquitinating enzyme CYLD (2019)
  • Author(s): Uematsu Atsushi、Kido Kohki、Takahashi Hirotaka、Takahashi Chikako、Yanagihara Yuta、Saeki Noritaka、Yoshida Shuhei、Maekawa Masashi、Honda Mamoru、Kai Tsutomu、Shimizu Kouhei、Higashiyama Shigeki、Imai Yuuki、Tokunaga Fuminori、Sawasaki Tatsuya
  • Journal Title: Journal of Biological Chemistry Volume: 294 Issue: 38 Pages: 14135-14148
  • DOI: 10.1074/jbc.ra119.010119
  • Peer Reviewed / Open Access
• [Journal Article] Identification of linear polyubiquitin chain immunoreactivity in tau pathology of Alzheimer’s disease (2019)
  • Author(s): Nakayama Yoshiaki、Sakamoto Shinji、Tsuji Kazumi、Ayaki Takashi、Tokunaga Fuminori、Ito Hidefumi
  • Journal Title: Neuroscience Letters Volume: 703 Pages: 53-57
  • DOI: 10.1016/j.neulet.2019.03.017
  • Peer Reviewed
• [Presentation] 大阪市立大学医学部1年生に対する臨床遺伝教育の試み (2020)
  • Author(s): 馬場遥香, 浄弘裕紀子, 堀田純子, 徳永文稔, 瀬戸俊之
  • Organizer: 第44回日本遺伝カウンセリング学会学術集会
• [Presentation] NDP52のユビキチン結合性を介した炎症応答・細胞死制御とLUBAC阻害剤の影響 (2020)
  • Author(s): 宮下裕久, 及川大輔, 寺脇正剛, 徳永文稔
  • Organizer: 第93回日本生化学会大会
• [Presentation] 非定型型直鎖状ユビキチン鎖を足場とするNF-κB活性化の細胞機能と数理モデル (2020)
  • Author(s): 徳永文稔, 及川大輔, 畑中尚也, 鈴木貴
  • Organizer: 第93回日本生化学会大会
  • Invited
• [Presentation] ヒトDUBアレイ技術を用いたUSP特異的阻害剤の開発 (2020)
  • Author(s): 高橋宏隆, 山中聡士, 桑田翔平, 檜垣佳奈, 佐藤裕介, 深井周也, 徳永文稔, 澤崎達也
  • Organizer: 第93回日本生化学会大会
• [Presentation] HOIPINsによるLUBAC活性抑制と自然免疫応答制御の分子基盤 (2020)
  • Author(s): 及川大輔, 徳永文稔
  • Organizer: 第93回日本生化学会大会
  • Invited
• [Presentation] cFLIPのユビキチン化による新たなアポトーシス抑制機構の解析 (2020)
  • Author(s): 中林修, 高橋宏隆, 村井晋, 大竹史明, 駒澤幸子, 土屋勇一, 佐伯泰, 吉田雪子, 山崎創, 徳永文稔, 森脇健太, 澤崎達也, 中野裕康
  • Organizer: 第93回日本生化学会大会
• [Presentation] Mathematical Model of Linear Ubiquitination-mediated NF-κB Activation Pathway (2020)
  • Author(s): Tokunaga F, Oikawa D, Hatanaka N, Suzuki T
  • Organizer: JSPS Core-to-Core Program “Establishing International Research Network of Mathematical Oncology”
  • Int'l Joint Research / Invited
• [Presentation] コムギ無細胞系を用いた直鎖状ユビキチン鎖の新規デコーダー分子の網羅的探索と機能解析 (2020)
  • Author(s): 高橋宏隆, 長尾和哉, 岩崎誠, 佐藤裕介, 及川大輔, 徳永文稔, 澤崎達也
  • Organizer: 第43回日本分子生物学会年会
  • Invited
• [Presentation] 非定型型直鎖状ユビキチン鎖を足場とした炎症シグナル発信とプロテイノパチー (2020)
  • Author(s): 徳永文稔, 伊東秀文, 及川大輔
  • Organizer: 第43回日本分子生物学会年会
  • Invited
• [Presentation] LRBA DUF1088ドメインの自然免疫制御における重要性 (2019)
  • Author(s): 及川大輔、駒倉啓大、福士雅也、入江崇、堀居拓郎、畑田出穂、阪口剛正、徳永文稔
  • Organizer: 第42回日本分子生物学会年会
• [Presentation] 直鎖状ポリユビキチン鎖結合タンパク質ZnUBPファミリーのNF-κB抑制機構の解明 (2019)
  • Author(s): 高橋宏隆、及川大輔、長尾和哉、岩崎誠、今井祐記、徳永文稔、澤崎達也
  • Organizer: 第42回日本分子生物学会年会
• [Presentation] 脱ユビキチン化酵素OTUD1によるNF-κB経路とインターフェロン産生経路の相反的制御 (2019)
  • Author(s): 駒倉啓大、及川大輔、徳永文稔
  • Organizer: 第42回日本分子生物学会年会
• [Presentation] 直鎖状ユビキチン鎖生成を介した炎症シグナル発信と疾患・創薬 (2019)
  • Author(s): 徳永文稔
  • Organizer: 第42回日本分子生物学会年会
  • Invited
• [Presentation] Hsp72によるがん細胞遊走制御 (2019)
  • Author(s): 塩田正之、田中昌子、鰐渕英機、徳永文稔
  • Organizer: 第14回日本臨床ストレス応答学会
• [Presentation] LUBACによるT細胞受容体シグナル制御に関する細胞・生化学及び数理モデル解析 (2019)
  • Author(s): 及川大輔、畑中尚也、鈴木貴、徳永文稔
  • Organizer: 第14回日本臨床ストレス応答学会
• [Presentation] Stress Signaling Modules: ストレスの受容と細胞応答の選択性 LUBACシグナルソームによる自然免疫応答の選択的制御 (2019)
  • Author(s): 及川 大輔, 駒倉 啓大, 阿部 貴則, 飯尾 清誠, 小比賀 真吾, 勝矢 健, 花田 和希, 坂本 信二, 徳永 文稔
  • Organizer: 第92回日本生化学会大会
• [Presentation] 自然免疫制御に関わる新規脱ユビキチン化酵素OTUD1の同定と生理機能解析 (2019)
  • Author(s): 駒倉啓大、徳永文稔
  • Organizer: 第24回日本病態プロテアーゼ学会学術集会
• [Presentation] Screening and characterization of novel LUBAC inhibitors, HOIPINs (2019)
  • Author(s): Oikawa, D., Katsuya, K., Hanada, K., Sugawara, K., Tsuruta, D., Sakamoto, S., Tokunaga, F.
  • Organizer: Cold Spring Harbor Laboratory Meeting on Ubiquitin, Autophagy & Disease
  • Int'l Joint Research
• [Remarks] 大阪市立大学大学院医学研究科分子病態学
  • URL: http://osaka-cu-1seika.umin.jp