Elucidation of the molecular mechanism involved in TLR7/8 response to ssRNA

• Project/Area Number: 16K08827
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Immunology
• Research Institution: The University of Tokyo
• Principal Investigator: Shibata Takuma 東京大学, 医科学研究所, 助教 (30554505)
• Research Collaborator: Sato Ryota ; Liu Kaiwen
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2018: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2017: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2016: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
• Keywords: Toll Like Receptor / TLR7 / TLR8 / ssRNA / Nucleoside / ヌクレオシド / マクロファージ / nuculeoside / 免疫学 / 核酸

Outline of Final Research Achievements

TLR7 in the endolysosome is a sensor for single-stranded RNA (ssRNA) fromviruses and it induces antiviral immune response. In addition, this receptor also responds to synthetic small molecules such as R848 and Imiquimod. However,it remains unclear how and why TLR7 can sense these two distinct ligands. We have found that TLR7 recognized guanosine (G),in the presence of uridine-containing oligoribonucleotide (U-ORN). With U-ORN, G/dG synergistically activated TLR7 and induced cytokine production in macrophages and pDCs. These results strongly suggest that TLR7 recognizes degradation products of ssRNA, G and U-ORN, but not ssRNA itself. Additionally, we have found that SLC29A3 deficient mice, which lose nucleoside transporter in endolysosomes, show autoinflammatory disease in TLR7-dependent manner. Our findings reveal that TLR7 works as a guanosine sensor and the appropriate control of nucleoside metabolism is indispensable for preventing excessive TLR7 response in vivo.

Academic Significance and Societal Importance of the Research Achievements

TLR7やTLR8は抗ウイルス応答の誘導に加えて自己免疫疾患や自己炎症性疾患の原因となっていることが以前から示唆されている。しかし、生体内においてTLR7/8の過剰応答が誘導される原因は良く分かっていなかった。本研究では、TLR7/8によるssRNA認識に核酸代謝関連遺伝子が関与することを証明すると共に、それら遺伝子のマウスにおける機能欠損が生体内でのTLR7/8過剰活性化を引き起こすことも示された。以上の事実は、核酸代謝関連遺伝子の機能異常がTLR7/8応答を介した病態の発症につながることを示唆しており、今後ヒトにおいてTLR7/8が関与した疾患の発見につながると期待される。

Research Products

• [Journal Article] Cytidine deaminase enables Toll-like receptor 8 activation by cytidine or its analogs. (2019)
  • Author(s): Furusho K, Shibata T, Sato R, Fukui R, Motoi Y, Zhang Y, Saitoh SI, Ichinohe T, Moriyama M, Nakamura S, Miyake, K.
  • Journal Title: International immunology. Volume: 31 Issue: 3 Pages: 167-173
  • DOI: 10.1093/intimm/dxy075
  • Peer Reviewed / Open Access
• [Journal Article] Structural Analyses of Toll-like Receptor 7 Reveal Detailed RNA Sequence Specificity and Recognition Mechanism of Agonistic Ligands (2018)
  • Author(s): Zhang Zhikuan、Ohto Umeharu、Shibata Takuma、Taoka Masato、Yamauchi Yoshio、Sato Ryota、Shukla Nikunj M.、David Sunil A.、Isobe Toshiaki、Miyake Kensuke、Shimizu Toshiyuki
  • Journal Title: Cell Reports Volume: 25 Issue: 12 Pages: 3371-3381
  • DOI: 10.1016/j.celrep.2018.11.081
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Combating herpesvirus encephalitis by potentiating a TLR3-mTORC2 axis. (2018)
  • Author(s): Sato R, Kato A, Chimura T, Saitoh SI, Shibata T, Murakami Y, Fukui R, Liu K, Zhang Y, Arii J, Sun-Wada GH, Wada Y, Ikenoue T, Barber GN, Manabe T, Kawaguchi Y, Miyake K.
  • Journal Title: Nat Immunol. Volume: 19 Issue: 10 Pages: 1071-1082
  • DOI: 10.1038/s41590-018-0203-2
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Structural basis for species-specific activation of mouse Toll-like receptor 9 (2018)
  • Author(s): Ishida Hanako、Ohto Umeharu、Shibata Takuma、Miyake Kensuke、Shimizu Toshiyuki
  • Journal Title: FEBS Letters Volume: 592 Issue: 15 Pages: 2636-2646
  • DOI: 10.1002/1873-3468.13176
  • Peer Reviewed / Open Access
• [Journal Article] Cleavage of Toll-Like Receptor 9 Ectodomain Is Required for In Vivo Responses to Single Strand DNA (2018)
  • Author(s): Fukui Ryutaro、Yamamoto Chikako、Matsumoto Fumi、Onji Masahiro、Shibata Takuma、Murakami Yusuke、Kanno Atsuo、Hayashi Takuto、Tanimura Natsuko、Yoshida Nobuaki、Miyake Kensuke
  • Journal Title: Frontiers in Immunology Volume: 9 Pages: 1491-1491
  • DOI: 10.3389/fimmu.2018.01491
  • Peer Reviewed / Open Access
• [Journal Article] Toll-like Receptor 9 Contains Two DNA Binding Sites that Function Cooperatively to Promote Receptor Dimerization and Activation (2018)
  • Author(s): Ohto Umeharu、Ishida Hanako、Shibata Takuma、Sato Ryota、Miyake Kensuke、Shimizu Toshiyuki
  • Journal Title: Immunity Volume: 48 Issue: 4 Pages: 649-658
  • DOI: 10.1016/j.immuni.2018.03.013
  • Peer Reviewed / Open Access
• [Journal Article] Mechanisms controlling nucleic acid-sensing Toll-like receptors (2018)
  • Author(s): Miyake Kensuke、Shibata Takuma、Ohto Umeharu、Shimizu Toshiyuki、Saitoh Shin-Ichiroh、Fukui Ryutaro、Murakami Yusuke
  • Journal Title: International Immunology Volume: 30 Issue: 2 Pages: 43-51
  • DOI: 10.1093/intimm/dxy016
  • Peer Reviewed / Open Access
• [Journal Article] The protective effect of the anti-Toll-like receptor 9 antibody against acute cytokine storm caused by immunostimulatory DNA (2017)
  • Author(s): Murakami Y, Fukui R, Motoi Y, Shibata T, Saitoh SI, Sato R, Miyake K.
  • Journal Title: Scientific Report Volume: 7 Issue: 1 Pages: 44042-44054
  • DOI: 10.1038/srep44042
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] TLR7 mediated viral recognition results in focal type I interferon secretion by dendritic cells (2017)
  • Author(s): Saitoh SI, Abe F, Kanno A, Tanimura N, Mori Saitoh Y, Fukui R, Shibata T, Sato K, Ichinohe T, Hayashi M, Kubota K, Kozuka-Hata H, Oyama M, Kikko Y, Katada T, Kontani K and Miyake K
  • Journal Title: Nat Commun Volume: 8 Issue: 1 Pages: 1592-1592
  • DOI: 10.1038/s41467-017-01687-x
  • Peer Reviewed / Open Access
• [Journal Article] Requirement of glycosylation machinery in TLR responses revealed by CRISPR/Cas9 screening (2017)
  • Author(s): Sato Ryota、Shibata Takuma、Tanaka Yu、Kato Chiharu、Yamaguchi Kiyoshi、Furukawa Yoichi、Shimizu Eigo、Yamaguchi Rui、Imoto Seiya、Miyano Satoru、Miyake Kensuke
  • Journal Title: International Immunology Volume: 29 Issue: 8 Pages: 347-355
  • DOI: 10.1093/intimm/dxx044
  • Peer Reviewed / Open Access
• [Journal Article] Emerging roles of the processing of nucleic acids and Toll-like receptors in innate immune responses to nucleic acids (2017)
  • Author(s): Miyake K, Shibata T, Ohto U, Shimizu T
  • Journal Title: J Leukoc Biol Volume: 101 Issue: 1 Pages: 135-142
  • DOI: 10.1189/jlb.4mr0316-108r
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Guanosine and its modified derivatives are endogenous ligands for TLR7 (2016)
  • Author(s): Shibata T, Ohto U, Nomura S, Kibata K, Motoi Y, Zhang Y, Murakami Y, Fukui R, Ishimoto T, Sano S, Ito T, Shimizu T, Miyake K.
  • Journal Title: Int Immunol Volume: - Issue: 5 Pages: 211-222
  • DOI: 10.1093/intimm/dxv062
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Structural Analysis Reveals that Toll-like Receptor 7 Is a Dual Receptor for Guanosine and Single-Stranded RNA (2016)
  • Author(s): Zhang Z, Ohto U, Shibata T, Krayukhina E, Taoka M, Yamauchi Y, Tanji H, Isobe T, Uchiyama S, Miyake K, Shimizu T
  • Journal Title: Immunity Volume: 45 Issue: 4 Pages: 737-748
  • DOI: 10.1016/j.immuni.2016.09.011
  • Peer Reviewed
• [Journal Article] Autoinhibition and relief mechanism by the proteolytic processing of Toll-like receptor 8. (2016)
  • Author(s): Tanji, H., Ohto, U., Motoi, Y., Shibata, T., Miyake, K., & Shimizu, T.
  • Journal Title: Proc. Natl. Acad. Sci. U S A Volume: 113 Issue: 11 Pages: 3012-3017
  • DOI: 10.1073/pnas.1516000113
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] TLR7によるグアノシン認識とその病態における意義 (2018)
  • Author(s): 柴田 琢磨
  • Organizer: 第39回 日本炎症・再生医学会 －炎症と再生の融合－
  • Invited
• [Presentation] Nucleosides are endogenous ligands for TLR7 and TLR8 (2017)
  • Author(s): Takuma Shibata
  • Organizer: Keystone Simposia, Viral Immunity: Mechanisms and Consequences (B4)
  • Place of Presentation: Hilton Santa Fe Historic Plaza Hotel , Santa Fe, New Mexico, USA
  • Year and Date: 2017-02-19
• [Presentation] Guanosine sensing by TLR7 and its implication in inflammatory disease (2017)
  • Author(s): Takuma Shibata
  • Organizer: 第46回 日本免疫学会学術集会
  • Int'l Joint Research / Invited
• [Presentation] Nucleosides are endogenous ligands for TLR7 and TLR8 (2017)
  • Author(s): Takuma Shibata
  • Organizer: Keystone symposium (Viral immunity)
  • Int'l Joint Research
• [Presentation] Guanosine and its analogs are endogenous ligands for TLR7 (2016)
  • Author(s): 柴田 琢磨
  • Organizer: 第39回 日本分子生物学会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2016-12-01
• [Remarks] 東京大学 医科学研究所 感染遺伝学分野
  • URL: http://www.ims.u-tokyo.ac.jp/kanseniden/
• [Remarks] 東京大学医科学研究所感染遺伝学分野
  • URL: http://www.ims.u-tokyo.ac.jp/kanseniden/
• [Patent(Industrial Property Rights)] SLC28又は29遺伝子異常症の治療剤又は予防剤 (2018)
  • Inventor(s): 三宅 健介、柴田 琢磨
  • Industrial Property Rights Holder: 三宅 健介、柴田 琢磨
  • Industrial Property Rights Type: 特許
  • Filing Date: 2018
• [Patent(Industrial Property Rights)] SLC28又は２９遺伝子異常症の治療剤又は予防剤 (2017)
  • Inventor(s): 三宅健介、柴田琢磨
  • Industrial Property Rights Holder: 三宅健介、柴田琢磨
  • Industrial Property Rights Type: 特許
  • Filing Date: 2017