Elucidation of the regulation mechanism of cystic fibrosis-specific splice variant expression and development of novel therapeutic strategies

• Project/Area Number: 19K07114
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 47040:Pharmacology-related
• Research Institution: Sojo University
• Principal Investigator: Shuto Keiko 崇城大学, 薬学部, 講師 (70510692)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2021: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2019: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: 嚢胞性線維症 / SIGIRR / スプライスバリアント / IL-37 / 化合物探索 / 嚢胞性線維症（CF） / HDAC3 / ヒストン脱アセチル化酵素（HDAC）阻害 / スプライススイッチ / ZIP2

Outline of Research at the Start

嚢胞性線維症（CF）は慢性気道炎症や粘液貯留を特徴とし、平均寿命約40歳の予後不良の遺伝性疾患である。これまでに申請者は、炎症制御や粘液産生に関与する２つの分子の変異体の発現がCF細胞特異的に上昇することを見出した。そこで本研究では、それら２つの分子の変異体発現調節メカニズムの解明と、変異体発現を絶対定量可能な評価系を用いて世界中の有用天然物由来の化合物を用いてスクリーニングを行い、新たな創薬に向けた基礎的検討を行う。

Outline of Final Research Achievements

Cystic Fibrosis (CF) is a hereditary disorder typically characterized by infection-associated chronic lung inflammation. Toll-like receptors (TLRs) are innate immune receptors that sense pathogen-associated molecular patterns (PAMPs) and their excessive or prolonged activation is considered as one of the mechanisms for CF hyperinflammatory phenotype. SIGIRR is a transmembrane protein acting as a negative regulator of multiple TLR signaling pathways after stimulation with anti-inflammatory cytokine IL-37, a ligand for SIGIRR.
We have recently demonstrated the defective surface expression of full-length SIGIRR (WT-SIGIRR) in CF airway epithelial cells due to its splicing variant Δ8-SIGIRR, suggesting impaired IL-37-depdendent anti-inflammatory activity.
In order to restore its surface expression in CF cells, we performed screening analysis and revealed that factor X inhibitor is a specific inducer for WT-SIGIRR.

Academic Significance and Societal Importance of the Research Achievements

CF気道上皮細胞おいて分子X阻害剤によりWT-SIGIRRを特異的に発現誘導する事は、分子X family阻害剤が持つ細胞毒性等の副作用を軽減しながらIL-37による抗炎症作用を回復させる為に有用である可能性が示唆された。
本知見は、抗炎症分子SIGIRRを強力に発現誘導する標的を明らかにした初めての報告であり、CF気道炎症に対する新たな治療戦略を提起するものである。

Research Products

• [Journal Article] Azithromycin Inhibits Constitutive Airway Epithelial Sodium Channel Activation <i>in Vitro</i> and Modulates Downstream Pathogenesis <i>in Vivo</i> (2020)
  • Author(s): Haruka Fujikawa, Taise Kawakami, Ryunosuke Nakashima, Aoi Nasu, Shunsuke Kamei, Hirofumi Nohara, Yuka Eto, Keiko Ueno-Shuto, Toru Takeo, Naomi Nakagata, Mary Ann Suico, Hirofumi Kai, Tsuyoshi Shuto.
  • Journal Title: Biological and Pharmaceutical Bulletin Volume: 43 Issue: 4 Pages: 725-730
  • DOI: 10.1248/bpb.b19-01091
  • NAID: 130007825345
  • ISSN: 0918-6158, 1347-5215
  • Year and Date: 2020-04-01
  • Peer Reviewed / Open Access
• [Journal Article] Higher Blood Uric Acid in Female Humans and Mice as a Protective Factor Against Pathophysiological Decline of Lung Function (2020)
  • Author(s): Fujikawa H, Sakamoto Y, Masuda N, Oniki K, Kamei S, Nohara H, Nakashima R, Maruta K, Kawakami T, Eto Y, Takahashi N, Takeo T, Nakagata N, Watanabe H, Otake K, Ogata Y, Tomioka NH, Hosoyamada M, Takada T, Ueno-Shuto K , Suico MA, Kai H, Saruwatari J, Shuto T.
  • Journal Title: Antioxidants Volume: 9 Issue: 5 Pages: 387-387
  • DOI: 10.3390/antiox9050387
  • Peer Reviewed / Open Access
• [Journal Article] Intratracheal GLP-1 receptor agonist treatment up-regulates mucin via p38 and exacerbates emphysematous phenotype in mucus hypersecretory obstructive lung diseases. (2020)
  • Author(s): Hirofumi Nohara, Ryunosuke Nakashima, Shunsuke Kamei, Haruka Fujikawa, KeikoUeno-Shuto, Taisei Kawakami, Yuka Eto, Mary Ann Suico, Jian-DongLi, Hirofumi Kai, Tsuyoshi Shuto.
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 524 Issue: 2 Pages: 332-339
  • DOI: 10.1016/j.bbrc.2020.01.081
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] The<I> DsbA-L</I> gene is associated with respiratory function of the elderly via its adiponectin multimeric or antioxidant properties. (2020)
  • Author(s): Kentaro Oniki, Hirofumi Nohara, Ryunosuke Nakashima, Yui Obata, Narumi Muto, Yuki Sakamoto, Keiko Ueno-Shuto, Tadashi Imafuku, Yu Ishima, Hiroshi Watanabe, Toru Maruyama, Koji Otake, Yasuhiro Ogata, Mary Ann Suico, Hirofumi Kai, Tsuyoshi Shuto, and Junji Saruwatari.
  • Journal Title: Scientific Reports Volume: 10 Issue: 1 Pages: 5973-5973
  • DOI: 10.1038/s41598-020-62872-5
  • Peer Reviewed / Open Access
• [Presentation] 正常および慢性閉塞性肺疾患モデルにおける抗ウイルス免疫抑制分子 PD-L1 の発現変動 (2021)
  • Author(s): 林恵、中嶋竜之介、高橋 宜暉、那須 葵、岸本 朋樹、首藤 恵子、Mary Ann Suico、甲斐 広文、首藤 剛
  • Organizer: 第38回日本薬学会九州山口支部大会