Treatment of sepsis with exosomal thrombomodulin

• Project/Area Number: 18K08917
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 55060:Emergency medicine-related
• Research Institution: Mie University
• Principal Investigator: KAWAMOTO EIJI 三重大学, 医学部附属病院, 講師 (20577415)
• Co-Investigator(Kenkyū-buntansha): 江口 暁子 三重大学, 医学系研究科, 特任講師(研究担当) (00598980) ; 島岡 要 三重大学, 医学系研究科, 教授 (40281133) ; 高娃 阿栄 三重大学, 医学系研究科, 助教 (50643805)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2020: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2019: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2018: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
• Keywords: トロンボモジュリン / 敗血症 / Fc融合蛋白 / エキソソーム / トロンボモジュリンFc蛋白 / ファイブロネクチン / 抗炎症作用 / ARDS / エキソソーム型トロンボモジュリン / リコモジュリン / CRISPR/CAS9 / dCas9活性化システム / SIRS / Sepsis / 細胞外小胞 / インテグリン / 遺伝子編集

Outline of Final Research Achievements

We created an extracellular domain of thrombomodulin, TMD123-Fc, or domain deletion TM-Fc proteins (TM domain 12-Fc, TM domain 23-Fc) and examined their bindings to fibronectin in vitro by ELISA. The lectin-like domain of thrombomodulin was found to be essential for the binding of the extracellular domain of thrombomodulin to fibronectin. Using a V-well cell ad-hesion assay or flow cytometry analysis with fluorescent beads, we found that both TMD123-Fc and TMD12-Fc inhibited the binding between β1 integrin of human breast cancer-derived cell lines and fibronectin. Furthermore, TMD123-Fc and TMD12-Fc inhibited the binding of activated integrins to fibronectin under shear stress in the presence of Ca2+ and Mg2+ but not under strong integrin-activation conditions in the presence of Mg2+ without Ca2+.

Academic Significance and Societal Importance of the Research Achievements

我々の知見は、炎症の比較的早い段階で外因性に投与されるthrom-bomodulin Fc融合タンパク質が、乳癌細胞株のβ1インテグリンとフィブロネクチンの結合を阻害する新しい治療法の開発に応用できることを示唆している。
本研究のようなTM-Fc型の安定化した蛋白の知見はエキソソーム型トロンボモジュリンの開発に役立てることができる。

Research Products

• [Journal Article] The Lectin-Like Domain of Thrombomodulin Inhibits β1 Integrin-Dependent Binding of Human Breast Cancer-Derived Cell Lines to Fibronectin. (2021)
  • Author(s): Kawamoto E, Nago N, Okamoto T, Gaowa A, Masui-Ito A, Akama Y, Darkwah S, Appiah MG, Myint PK, Obeng G, Ito A, Caidengbate S, Esumi R, Yamaguchi T, Park EJ, Imai H, Shimaoka M.
  • Journal Title: Biomedicines Volume: 9 Issue: 2 Pages: 162-162
  • DOI: 10.3390/biomedicines9020162
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Ligand-competent fractalkine receptor is expressed on exosomes. (2021)
  • Author(s): Park EJ, Myint PK, Appiah MG, Worawattananutai P, Inprasit J, Prajuabjinda O, Soe ZY, Gaowa A, Kawamoto E, Shimaoka M.
  • Journal Title: Biochem Biophys Rep. Volume: 26 Pages: 100932-100932
  • DOI: 10.1016/j.bbrep.2021.100932
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] The Spike Glycoprotein of SARS-CoV-2 Binds to β1 Integrins Expressed on the Surface of Lung Epithelial Cells. (2021)
  • Author(s): Park EJ, Myint PK, Appiah MG, Darkwah S, Caidengbate S, Ito A, Matsuo E, Gaowa a, Shimaoka M. Kawamoto E,
  • Journal Title: Viruses Volume: 13 Issue: 4 Pages: 645-645
  • DOI: 10.3390/v13040645
  • Peer Reviewed / Open Access
• [Journal Article] Irisin supports integrin-mediated cell adhesion of lymphocytes. (2021)
  • Author(s): Myint PK, Ito A, Appiah MG, Obeng G, Darkwah S, Kawamoto E, Gaowa A, Park EJ, Shimaoka M
  • Journal Title: Biochem Biophys Rep. Volume: 26 Pages: 100977-100977
  • DOI: 10.1016/j.bbrep.2021.100977
  • Peer Reviewed / Open Access
• [Journal Article] Remodeling of Bone Marrow Niches and Roles of Exosomes in Leukemia. (2021)
  • Author(s): Yamaguchi T, Kawamoto E, Gaowa A, Park EJ, Shimaoka M.
  • Journal Title: Int J Mol Sci. Volume: 22 Issue: 4 Pages: 1881-1881
  • DOI: 10.3390/ijms22041881
  • Peer Reviewed / Open Access
• [Journal Article] Potential Roles of Muscle-Derived Extracellular Vesicles in Remodeling Cellular Microenvironment: Proposed Implications of the Exercise-Induced Myokine, Irisin (2021)
  • Author(s): Darkwah Samuel、Park Eun Jeong、Myint Phyoe Kyawe、Ito Atsushi、Appiah Michael G.、Obeng Gideon、Kawamoto Eiji、Shimaoka Motomu
  • Journal Title: Frontiers in Cell and Developmental Biology Volume: 9 Pages: 1-1
  • DOI: 10.3389/fcell.2021.634853
  • Peer Reviewed / Open Access
• [Journal Article] Sepsis Induces Deregulation of IL-13 Production and PD-1 Expression in Lung Group 2 Innate Lymphoid Cells. (2021)
  • Author(s): Akama Y, Park EJ, Satoh-Takayama N, Gaowa A, Ito A, Kawamoto E, Darkwah S, Appiah MG, Myint PK, Ohno H, Imai H, Shimaoka M.
  • Journal Title: Shock Volume: 55 Issue: 3 Pages: 357-370
  • DOI: 10.1097/shk.0000000000001647
  • Peer Reviewed / Open Access
• [Journal Article] How ICU Patient Severity Affects Communicative Interactions Between Healthcare Professionals: A Study Utilizing Wearable Sociometric Badges (2020)
  • Author(s): Kawamoto Eiji、Ito-Masui Asami、Esumi Ryo、Imai Hiroshi、Shimaoka Motomu
  • Journal Title: Frontiers in Medicine Volume: 7 Pages: 606987-606987
  • DOI: 10.3389/fmed.2020.606987
  • Peer Reviewed / Open Access
• [Journal Article] Methods to Study Integrin Functions on Exosomes (2020)
  • Author(s): Kawamoto Eiji、Park Eun Jeong、Shimaoka Motomu
  • Journal Title: Methods in Molecular Biology Volume: 2217 Pages: 265-281
  • DOI: 10.1007/978-1-0716-0962-0_15
  • ISBN: 9781071609613, 9781071609620
  • Peer Reviewed / Open Access
• [Journal Article] Integrin-Ligand Interactions in Inflammation, Cancer, and Metabolic Disease: Insights Into the Multifaceted Roles of an Emerging Ligand Irisin (2020)
  • Author(s): Park Eun Jeong、Myint Phyoe Kyawe、Ito Atsushi、Appiah Michael G.、Darkwah Samuel、Kawamoto Eiji、Shimaoka Motomu
  • Journal Title: Frontiers in Cell and Developmental Biology Volume: 8 Pages: 588066-588066
  • DOI: 10.3389/fcell.2020.588066
  • Peer Reviewed / Open Access
• [Journal Article] Intestinal Epithelium-Derived Luminally Released Extracellular Vesicles in Sepsis Exhibit the Ability to Suppress TNF-a and IL-17A Expression in Mucosal Inflammation. (2020)
  • Author(s): Appiah MG, Park EJ, Darkwah S, Kawamoto E, Akama Y, Gaowa A, Kalsan M, Ahmad S, Shimaoka M.
  • Journal Title: Int J Mol Sci. Volume: 21 Issue: 22 Pages: 8445-8445
  • DOI: 10.3390/ijms21228445
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] The Role of Innate Lymphoid Cells in the Regulation of Immune Homeostasis in Sepsis-Mediated Lung Inflammation. (2020)
  • Author(s): Akama Y, Park EJ, Satoh-Takayama N, Gaowa A, Ito A, Kawamoto E, Darkwah S, Appiah MG, Myint PK, Ohno H, Imai H, Shimaoka M.
  • Journal Title: Diagnostics (Basel) Volume: 10 Issue: 10 Pages: 808-808
  • DOI: 10.3390/diagnostics10100808
  • NAID: 120006941006
  • Peer Reviewed / Open Access
• [Journal Article] Recombinant Human Soluble Thrombomodulin Suppresses Monocyte Adhesion by Reducing Lipopolysaccharide-Induced Endothelial Cellular Stiffening (2020)
  • Author(s): Okamoto Takayuki、Kawamoto Eiji、Usuda Haruki、Tanaka Tetsuya、Nikai Tetsuro、Asanuma Kunihiro、Suzuki Koji、Shimaoka Motomu、Wada Koichiro
  • Journal Title: Cells Volume: 9 Issue: 8 Pages: 1811-1811
  • DOI: 10.3390/cells9081811
  • Peer Reviewed / Open Access
• [Journal Article] Targeted remodeling of breast cancer and immune cell homing niches by exosomal integrins. (2020)
  • Author(s): Myint PK, Park EJ, Gaowa A, Kawamoto E, Shimaoka M.
  • Journal Title: Diagn Pathol. Volume: 15 Issue: 1 Pages: 38-38
  • DOI: 10.1186/s13000-020-00959-3
  • Peer Reviewed / Open Access
• [Journal Article] Virtual reality as a non-pharmacologic analgesic for fasciotomy wound infections in acute compartment syndrome: a case report (2020)
  • Author(s): Esumi Ryo、Yokochi Ayumu、Shimaoka Motomu、Kawamoto Eiji
  • Journal Title: Journal of Medical Case Reports Volume: 14 Issue: 1 Pages: 46-46
  • DOI: 10.1186/s13256-020-02370-4
  • Peer Reviewed / Open Access
• [Journal Article] Feasibility of Measuring Face-to-Face Interactions among ICU Healthcare Professionals Using Wearable Sociometric Badges (2020)
  • Author(s): Ito-Masui Asami、Kawamoto Eiji、Nagai Yuki、Takagi Yuto、Ito Mami、Mizutani Noriko、Yano Ken’ichi、Yano Kazuo、Imai Hiroshi、Shimaoka Motomu
  • Journal Title: American Journal of Respiratory and Critical Care Medicine Volume: 201 Issue: 2 Pages: 245-247
  • DOI: 10.1164/rccm.201904-0779le
  • Peer Reviewed / Open Access
• [Journal Article] Exosomes in Sepsis and Inflammatory Tissue Injury (2019)
  • Author(s): Park EJ, Appiah MG, Myint PK, Gaowa A, 他2名
  • Journal Title: Curr Pharm Des Volume: 25 Issue: 42 Pages: 4486-4495
  • DOI: 10.2174/1381612825666191116125525
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Differential Roles of Dendritic Cells in Expanding CD4 T Cells in Sepsis (2019)
  • Author(s): Darkwah Samuel、Nago Nodoka、Appiah Michael G.、Myint Phyoe Kyawe、Kawamoto Eiji、Shimaoka Motomu、Park Eun Jeong
  • Journal Title: Biomedicines Volume: 7 Issue: 3 Pages: 52-52
  • DOI: 10.3390/biomedicines7030052
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Talin-2 regulates integrin functions in exosomes (2019)
  • Author(s): Soe Zay Yar、Prajuabjinda Onmanee、Myint Phyoe Kyawe、Gaowa Arong、Kawamoto Eiji、Park Eun Jeong、Shimaoka Motomu
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 512 Issue: 3 Pages: 429-434
  • DOI: 10.1016/j.bbrc.2019.03.027
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Anti-adhesive effects of human soluble thrombomodulin and its domains (2019)
  • Author(s): Kawamoto Eiji、Nago Nodoka、Okamoto Takayuki、Gaowa Arong、Masui-Ito Asami、Sakakura Yosuke、Akama Yuichi、Soe Zay Yar、Prajuabjinda Onmanee、Darkwah Samuel、Appiah Michael G.、Myint Phyoe Kyawe、Obeng Gideon、Park Eun Jeong、Imai Hiroshi、Shimaoka Motomu
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 511 Issue: 2 Pages: 312-317
  • DOI: 10.1016/j.bbrc.2019.02.041
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Connexins and Integrins in Exosomes (2019)
  • Author(s): Shimaoka M, Kawamoto E, Gaowa A, Okamoto T, Park EJ
  • Journal Title: Cancers (Basel) Volume: － Issue: 1 Pages: 106-106
  • DOI: 10.3390/cancers11010106
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Connexins and Integrins in Exosomes. (2019)
  • Author(s): Shimaoka M, Kawamoto E, Gaowa A, Okamoto T, Park EJ.
  • Journal Title: Cancers (Basel). Volume: 11(1)
  • Peer Reviewed / Open Access
• [Journal Article] Integrin and PD-1 Ligand Expression on Circulating Extracellular Vesicles in Systemic Inflammatory Response Syndrome and Sepsis (2018)
  • Author(s): Kawamoto Eiji、Masui-Ito Asami、Eguchi Akiko、Soe Zay Yar、Prajuabjinda Onmanee、Darkwah Samuel、Park Eun Jeong、Imai Hiroshi、Shimaoka Motomu
  • Journal Title: SHOCK Volume: - Issue: 1 Pages: 1-1
  • DOI: 10.1097/shk.0000000000001228
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Exosomal regulation of lymphocyte homing to the gut (2018)
  • Author(s): Park Eun Jeong、Prajuabjinda Onmanee、Soe Zay Yar、Darkwah Samuel、Appiah Michael G.、Kawamoto Eiji、Momose Fumiyasu、Shiku Hiroshi、Shimaoka Motomu
  • Journal Title: Blood Advances Volume: 3 Issue: 1 Pages: 1-11
  • DOI: 10.1182/bloodadvances.2018024877
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Biomechanical profiles of tracheal intubation: a mannequin-based study to make an objective assessment of clinical skills by expert anesthesiologists and novice residents. (2018)
  • Author(s): Sakakura Y, Kamei M, Sakamoto R, Morii H, Itoh-Masui A, Kawamoto E, Imai H, Miyabe M, Shimaoka M.
  • Journal Title: BMC Med Educ Volume: Dec 4;18(1) Issue: 1 Pages: 293-293
  • DOI: 10.1186/s12909-018-1410-0
  • Peer Reviewed / Open Access
• [Journal Article] Reduced substrate stiffness promotes M2-like macrophage activation and enhances peroxisome proliferator-activated receptor-gamma expression (2018)
  • Author(s): Okamoto T, Takagi Y, Kawamoto E, Park EJ, Usuda H, Wada K, Shimaoka M
  • Journal Title: Experimental Cell Research Volume: 367 Issue: 2 Pages: 264-273
  • DOI: 10.1016/j.yexcr.2018.04.005
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] 血漿中PD-1リガンドと敗血症患者の臨床経過や臓器障害との相関の検討 (2019)
  • Author(s): 川本 英嗣, 伊藤 亜紗実, 江口 暁子, Zay Yar Soe, Prajuabjinda Onmanee, Darkwah Samuel, 朴 恩正, 今井 寛, 島岡 要
  • Organizer: 日本血栓止血学会
• [Presentation] Integrin and PD-1 Ligand Expression on Circulating Extracellular Vesicles in Systemic Inflammatory Response Syndrome and Sepsis (2018)
  • Author(s): Eiji Kawamoto
  • Organizer: 41 st ANNUAL CONFERENCE ON SHOCK
  • Int'l Joint Research
• [Presentation] 細胞外小胞（EV）インテグリンの敗血症病態への関与の研究を目的としたCRISPR/Cas9遺伝子編集技術を用いたインテグリンノックアウトEVがそれぞれSIRSの作製 (2018)
  • Author(s): 川本英嗣
  • Organizer: 第33回日本ショック学会総会