肺における細胞特異的エピジェネティクスへの挑戦

2009 Fiscal Year Annual Research Report

• Project/Area Number: 21590961
• Research Institution: Hokkaido University
• Principal Investigator: 別役 智子 Hokkaido University, 大学院・医学研究科, 准教授 (60333605)
• Keywords: LCM / マウス / 喫煙 / 細気管支上皮 / miRNA

Research Abstract

炎症性サイトカインの調節、DNA修復、増殖といった様々な細胞機能の変化は、いわゆるエピジェネティクスと呼ばれるDNAの翻訳後修飾によりもたらされている。肺におけるmiRNAに関する既存の報告はごく限られている。よって、本研究をこれらのLCMを用いたジェネティクス研究成果のエピジェネティクスへの発展的展開として位置づける。本年度は、喫煙暴露後とコントロールマウスとの間で細気管支上皮におけるmiRNAをマイクロアレイ上で網羅的にスクリーニングすることを目的とした。C57BL/6マウスに喫煙暴露モデルを作成する。laser capture microdissection(LCM)法を用いて顕微鏡下に細気管支上皮細胞を選択的に採取した。長さ200塩基以下のRNAを濃縮したsmall RNAの画分として回収できるように開発されたmirVana? miRNA Isolatiom Kit(Ambion社)を用いた。miRNAはendonucleaseによるプロセッシングの結果保存された部分を有している。マウスの既知miRNAの数が比較的少ないことにより、すべてのmiRNAを理想的にアレイというプラットフォーム上で拾い上げることができ、既存のバイオインフォーマティクスのデータベースの中に取り込んで解析を行った。

Research Products

• [Journal Article] Cigarette smoke induces nucleic acid oxidation in lung fibroblasts (2010)
  • Author(s): Deslee G, Adair-Kirk TL, Betsuyaku T, Woods JC, Moore CH, Gierada DS, Conradi SH, Atkinson JJ, Toennies HM, Battaile JT, Kobayashi DK, Patterson GA, Holtzman MJ, Pierce RA
  • Journal Title: Am J Respir Cell Mol Biol (in press)
  • Peer Reviewed
• [Journal Article] High levels of extracellular matrix metalloproteinase inducer are expressed in lymphangioleiomyomatosis (2010)
  • Author(s): Odajima N, Betsuyaku T, Yoshida T, Hosokawa T, Nishimura M
  • Journal Title: Hum Pathol (in press)
  • Peer Reviewed
• [Journal Article] Aging enhances susceptibility to cigarette smoke-induced inflammation through bronchiolar chemokines (2010)
  • Author(s): Moriyama C, Betsuyaku T, Ito Y, Hamamura I, Hata J, Takahashi H, Nasuhara Y, Nishimura M
  • Journal Title: Am J Respir Cell Mol Biol 42 Pages: 304-311
  • Peer Reviewed
• [Journal Article] Laminin gamma2 fragments are increased in the circulation of patients with early phase acute lung injury (2010)
  • Author(s): Katayama M, Ishizaka A, Sakamoto M, Fujishima S, Sekiguchi K, Asano K, Betsuyaku T, Kotani T, Ware LB, Matthay MA, Hashimoto S
  • Journal Title: Intensice Care Med 36 Pages: 479-486
  • Peer Reviewed
• [Journal Article] Aging affects lipopolysaccharide-induced upregulation of heme oxygenase-1 in the lungs and alveolar macrophages (2009)
  • Author(s): Ito Y, Betsuyaku T, Moriyama C, Nasuhara Y, Nishimura M
  • Journal Title: Biogerontology 10 Pages: 173-180
  • Peer Reviewed
• [Journal Article] Matrix metalloproteinases in blood from patients with LAM (2009)
  • Author(s): Odajima N, Betsuyaku T, Nasuhara Y, Inoue H, Seyama K, Nishimura M
  • Journal Title: Respir Med 103 Pages: 124-129
  • Peer Reviewed
• [Journal Article] A soluble factor (EMMPRIN) in exudates influences knee motion after total arthroplasty (2009)
  • Author(s): Onodera J, Onodera S, Kondo E, Betsuyaku T, Yasuda K
  • Journal Title: Knee Surg Sports Traumatol Arthrosc 17 Pages: 298-304
  • Peer Reviewed
• [Journal Article] Relationship between improved airflow limitation and changes in airway caliber induced by inhaled anticholinergic agents in COPD (2009)
  • Author(s): Hasegawa M, Makita H, Nasuhara Y, Odajima N, Nagai K, Ito Y, Betsuyaku T, Nishimura M
  • Journal Title: Thorax 64 Pages: 332-338
  • Peer Reviewed
• [Journal Article] Curcumin attenuates elastase-and cigarette smoke-induced pulmonary emphysema in mice (2009)
  • Author(s): Suzuki M, Betsuyaku T, Ito Y, Nagai K, Odajima N, Moriyama C, Nasuhara Y, Nishimura M
  • Journal Title: Am J Physiol Lung Cell Mol Physiol 296 Pages: L614-L623
  • Peer Reviewed
• [Journal Article] Epilysin(MMP-28)restrains early macrophage recruitment in Pseudomonas aeruginosa pneumonia (2009)
  • Author(s): Manicone AM, Birkland TP, Lin M, Betsuyaku T, van Rooijen N, Lohi J, Keski-Oja J, Wang Y, Skerrett SJ, Parks WC
  • Journal Title: J Immunol 182 Pages: 3866-3876
  • Peer Reviewed
• [Presentation] Aging enhances susceptibility to cigarette smoke-induced inflammation (2009)
  • Author(s): Betsuyaku T
  • Organizer: The 9th World Congress on Inflammation Industry Sponsored Symposium 1
  • Place of Presentation: 東京:京王プラザホテル東京
  • Year and Date: 2009-07-07
• [Presentation] Relationship between improved airflow limitation and changes in airway caliber induced by inhaled anticholinergics in chronic obstructive pulmonary disease (2009)
  • Author(s): Hasegawa M, Nasuhara Y, Makita H, Odajima N, Nagai K, Ito Y, Betsuyaku T, Nishimura M
  • Organizer: 第49回日本呼吸器学会
  • Place of Presentation: 東京:東京国際フォーラム
  • Year and Date: 2009-06-14
• [Presentation] Kinetics of various ocidative stress markers in bronchoalveolar lavage fluid (BALF) in lipopolysaccaride (LPS) induced lung injury (2009)
  • Author(s): Yoshida T, Betsuyaku T, Nagai K, Ito Y, Nasuhara Y, Nishimura M
  • Organizer: 第49回日本呼吸器学会
  • Place of Presentation: 東京:東京国際フォーラム
  • Year and Date: 2009-06-13
• [Presentation] Catalase expression in pulmonary macrophages of smokers and chronic obstructive pulmonary disease (COPD) patients (2009)
  • Author(s): Inomata T, Betsuyaku T, Ito Y, Nasuhara Y, Nishimura M
  • Organizer: 第49回日本呼吸器学会
  • Place of Presentation: 東京:東京国際フォーラム
  • Year and Date: 2009-06-13
• [Presentation] 酸化ストレスによる肺病変としての気腫化と線維化 (2009)
  • Author(s): 別役智子
  • Organizer: 第49回日本呼吸器学会
  • Place of Presentation: 東京:東京国際フォーラム
  • Year and Date: 2009-06-13
• [Presentation] アレルギー性気道炎症モデルにおける気道被覆液中のグルタチオン化蛋白の検討 (2009)
  • Author(s): 長井桂,別役智子,南須原康行,近藤宇史,西村正治
  • Organizer: 第49回日本呼吸器学会
  • Place of Presentation: 東京:東京国際フォーラム
  • Year and Date: 2009-06-13
• [Presentation] 高齢マウス単回喫煙曝露モデルにおける細気管支上皮細胞でのnuclear factor κB(NF-κB)の活性化とケモカイン発現亢進 (2009)
  • Author(s): 守山千夏,別役智子,伊藤洋子,西村正治
  • Organizer: 第49回日本呼吸器学会
  • Place of Presentation: 東京:東京国際フォーラム
  • Year and Date: 2009-06-12
• [Presentation] Alveolar-specific gene expression profiling in emphysematous lungs (2009)
  • Author(s): Hersh CP, Betsuyaku T, Patel N, Marchetti N, Klanderman BJ, Criner G, Choi A
  • Organizer: The 105^<th> American Thoracic Society International Conference
  • Place of Presentation: The San Diego Convention Center, San Diego, CA, United States
  • Year and Date: 2009-05-17
• [Presentation] Protein S-glutathionylation in the Epithelial Lining Fluid during Allergic Airway Inflammation (2009)
  • Author(s): Nagai K, Betsuyaku T,Nasuhara Y, Kondo T, Nishimura M
  • Organizer: The 105^<th> American Thoracic Society International Conference
  • Place of Presentation: The San Diego Convention Center, San Diego, CA, United States
  • Year and Date: 2009-05-17
• [Presentation] Duration of cigarette smoke (CS) exposure affects the resolution of lung inflammation after cessation in mice (2009)
  • Author(s): Odajima N, Betsuyaku T, Inomata T, Yoshida T, Moriyama C, Suzuki K, Nagai K, Nasuhara Y, Nishimura M
  • Organizer: The 105^<th> American Thoracic Society International Conference
  • Place of Presentation: The San Diego Convention Center, San Diego, CA, United States
  • Year and Date: 2009-05-17
• [Presentation] Kinetics of various oxidative stress markers in BAL fluid in lipopolysaccaride (LPS)-induced lung injury (2009)
  • Author(s): Yoshida T, Betsuyaku T, Nagai K, Ito Y, Nasuhara Y, Nishimura M
  • Organizer: The 105^<th> American Thoracic Society International Conference
  • Place of Presentation: The San Diego Convention Center, San Diego, CA, United States
  • Year and Date: 2009-05-17
• [Presentation] Catalase expression in pulmonary macrophages of smokers and chronic obstructive pulmonary disease(COPD)patients (2009)
  • Author(s): Inomata T, Betsuyaku T, Ito Y, Nasuhara Y, Nishimura M
  • Organizer: The 105^<th> American Thoracic Society International Conference
  • Place of Presentation: The San Diego Convention Center, San Diego, CA, United States
  • Year and Date: 2009-05-17
• [Book] 実験医学増刊 病態解明に迫るI.活性酸素シグナルと酸化ストレス 癌,神経変性疾患,循環・代謝異常にかかわるレドックス制御機構と最新の技術開発 (2009)
  • Author(s): 別役智子
  • Total Pages: 243
  • Publisher: 羊土社