Investigation of safer methods of therapeutic miRNA and exosomes

• Project/Area Number: 16K09610
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Kidney internal medicine
• Research Institution: Nagoya University
• Principal Investigator: Kato Noritoshi 名古屋大学, 医学部附属病院, 病院助教 (90716052)
• Co-Investigator(Kenkyū-buntansha): 丸山 彰一 名古屋大学, 医学系研究科, 教授 (10362253) ; 小杉 智規 名古屋大学, 医学系研究科, 講師 (90584681)
• Research Collaborator: Nishio Fumitoshi ; Funahashi Yoshio
• 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: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2017: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2016: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: miRNA / Toll like receptor / 敗血症 / microRNA / AKI / 医療・福祉

Outline of Final Research Achievements

We investigated the pathophysiological role of exogenously applied microRNA (miRNA) in sepsis-induced multiple organ injury. In vitro, we tested possible miRNAs which suppressed the production of pro-inflammatory cytokines. Of these, miR-146a displayed the highest suppressive effect. Sepsis was induced in mice via cecal ligation and puncture (CLP) and an intravenous injection of a complex of miR-146a-expressing plasmid and polyethyleneimine. Treatment with this complex significantly decreased the level of serum inflammatory cytokines, attenuated organ injury, and led to increased survival from sepsis. miR-146a-expressing plasmid was abundantly distributed in splenic macrophages. CLP mice treated with miR-146a displayed significantly decreased NF-κB activation in the spleen. The collective results support the conclusion that the induction of miR-146a expression in splenic macrophages prevents excessive inflammation and sepsis-induced multiple organ injury.

Academic Significance and Societal Importance of the Research Achievements

敗血症は全死亡率が高い重篤な疾患であるが、抗菌治療や、補液などに治療的なエビデンスがあるものの、新たな治療法の開発は遅れ、生存率の改善は停滞している。我々はToll like receptorシグナルをmiRNAによって制御するといった、全く新しいアプローチで治療を行った。
結果から全身投与したmiRNA発現ベクターは、主に脾臓マクロファージに取り込まれ、過剰なサイトカイン産生を抑制し、生存率の改善に寄与していた。これは今後の核酸医薬の開発にとって、投与核酸の体内動態、作用を探る上でも意義深い。また、miRNAは低分子医薬や抗体医薬と比べ安価に作成できるため、将来の治療応用にも期待が持てる。

Research Products

• [Journal Article] miR-146a targeted to splenic macrophages prevents sepsis-induced multiple organ injury. (2019)
  • Author(s): Funahashi Y, Kato N, Masuda T, Nishio F, Kitai H, Ishimoto T, Kosugi T, Tsuboi N, Matsuda N, Maruyama S, Kadomatsu K.
  • Journal Title: Lab. Invest. Volume: - Issue: 8 Pages: 1130-1142
  • DOI: 10.1038/s41374-019-0190-4
  • Peer Reviewed / Int'l Joint Research
• [Presentation] miR-146a Targeting the Splenic Macrophages Prevents Sepsis-Induced Acute Kidney Injury (2018)
  • Author(s): Funahashi Y, Kato N, Masuda T, Nishio F, Kitai H, Ishimoto T, Kosugi T, Tsuboi N, Matsuda N, Maruyama S, Kadomatsu K
  • Organizer: 41st Annual Conference on Shock
  • Int'l Joint Research
• [Presentation] miR-146a targeting the splenic macrophages prevents sepsis-induced acute kidney injury. (2018)
  • Author(s): Funahashi Y, Kato N, Kitai H, Ishimoto T, Kosugi T, Tsuboi N, Maruyama S, Kadomatsu K
  • Organizer: ISN Frontiers Meeting
  • Int'l Joint Research
• [Presentation] Yoshio Funahashi (2018)
  • Author(s): miR-146a Targeting the Spleen Prevents Sepsis-Induced Acute Kidney Injury
  • Organizer: ISN Frontiers Meeting 2018
  • Int'l Joint Research
• [Presentation] INDUCTION OF IMMUNOSUPPRESSIVE MICRO-RNA IN SPLEEN ATTENUATES SEPSIS INDUCED ACUTE KIDNEY INJURY (2017)
  • Author(s): Yoshio Funahashi
  • Organizer: 54th ERA-EDTA Congress
  • Int'l Joint Research
• [Presentation] Indirect therapeutic role of immunosuppressive micro-RNA for sepsis induced acute kidney injury via spleen. (2017)
  • Author(s): Yoshio Funahashi
  • Organizer: Kidney Week 2017
  • Int'l Joint Research
• [Presentation] 心腎連関における、活性化腎線維芽細胞由来エクソソームが持つ役割の検討 The potential roles of exosomes from activated kidney fibroblast on cardio-renal syndrome. (2016)
  • Author(s): 加藤規利、西尾文利、船橋嘉夫、丸山彰一
  • Organizer: 第8回日本RNAi研究会 第3回日本細胞外小胞学会
  • Place of Presentation: グランドプリンスホテル広島
  • Year and Date: 2016-09-01
• [Presentation] Exosomes from activated kidney fibroblast have ambivalent potential effect on progression of atherosclerosis. (2016)
  • Author(s): Fumitoshi Nishio, Noritoshi Kato, Takuji Ishimoto, Tomoki Kosugi, Naotake Tsuboi, Shoichi Maruyama, Seiichi Matsuo.
  • Organizer: 53RD ERA-EDTA CONGRESS
  • Place of Presentation: Vienna, Austria
  • Year and Date: 2016-05-22
  • Int'l Joint Research