microRNAの制御による動脈硬化症の治療法開発

• Project/Area Number: 20659146
• Research Category: Grant-in-Aid for Exploratory Research
• Allocation Type: Single-year Grants
• Research Field: Metabolomics
• Research Institution: Kyoto University
• Principal Investigator: 北 徹 Kyoto University, 医学研究科, 教授 (60161460)
• Co-Investigator(Kenkyū-buntansha): 尾野 亘 京都大学, 医学研究科, 助教 (00359275)
• Project Period (FY): 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥3,200,000 (Direct Cost: ¥3,200,000); Fiscal Year 2008: ¥3,200,000 (Direct Cost: ¥3,200,000)
• Keywords: microRNA / 動脈硬化 / 脂肪細胞 / ウイルス

Research Abstract

虚血性心疾患や脳卒中、動脈瘤などの粥状動脈硬化に起因する疾患は、悪性腫瘍と並ぶ我が国の2大死亡原因であり、粥状動脈硬化性疾患の成因解明や有効な予防・治療法の開発はこれからの高齢社会において火急の課題である。
申請者らは粥状動脈硬化の分子・細胞・個体レベルでの解明に一貫して取り組んできた。今回我々は、血管のみならず、脂肪組織、肝臓を含めた代謝制御に関わるmiRNAの機能の制御が、動脈硬化の治療に応用できないかどうか検討を始めた。まず、脂肪細胞、肝細胞、血管内皮細胞、血管平滑筋細胞において、miRNA合成を抑制したところ(レンチウイルスによるDicer、Ago2 siRNA発現)、細胞のATPが低下し、細胞死が亢進した。さらに、脂肪細胞、肝細胞、血管内皮細胞、血管平滑筋細胞特異的miRNAの発現ライブラリーを構築した。
次に、高脂肪食負荷マウスモデルの内臓脂肪、肝臓においてmiRNAの発現プロファイル解析を行ったところ、脂肪細胞分化に必要とされるmiR-143の発現が増加し、その標的遺伝子ERK5の発現減少を認めた。レンチウイルスを用いてmiR-143のデコイ遺伝子を脂肪組織で発現させ、miR-143を抑制するための予備実験として、レンチウイルスを用いて脂肪組織に特異的にGFP遺伝子を導入できることを確認した。現在、miR-143のデコイ遺伝子のマウスへの導入を行ったところである。このことにより、高脂肪食負荷での肥満を改善出来るかどうかを確認する。さらに動脈硬化モデルマウスであるApoE欠損マウスの動脈硬化巣より、miRNA発現プロファイル解析用のRNAを抽出した。

Research Products

• [Journal Article] TG-interacting factor is required for the differentiation of preadipocytes. (2008)
  • Author(s): Horie-T, Ono-K, Kinoshita-M, Nishi-H, Nagao-K, Kawamura-T, Abe-Y, Wada-H, Shimatsu-A, Kita-T, Hasegawa-K.
  • Journal Title: J Lipid Res. 49 Pages: 1224-34
  • Peer Reviewed
• [Journal Article] Oxidative stress induces GLUT4 translocation by activation of PI3-K/Akt and dual AMPK kinase in cardiac myocytes. (2008)
  • Author(s): Horie-T, Ono-K, Nagao-K, Nishi-H, Kinoshita-M, Kawamura-T, Wada-H, Shimatsu-A, Kita-T, Hasegawa-K.
  • Journal Title: J Cell Physiol. 215 Pages: 733-42
  • Peer Reviewed
• [Journal Article] Up-regulated expression of microRNA-143 in association with obesity in adipose tissue of mice fed high-fat diet. (2008)
  • Author(s): Takanabe R, Ono K, Abe Y, Takaya T, Horie T, Wada H, Kita T, Satoh N, Shimatsu A, Hasegawa K.
  • Journal Title: Biochem Biophys Res Commun. 376 Pages: 728-32
  • Peer Reviewed
• [Journal Article] Inhibition of CCN1 suppressed neointimal hyperplasia in a rat balloon injury model. (2008)
  • Author(s): Matsumae-H, Yoshida-Y, Ono-K, Togi-K, Inoue-K, Furukawa-Y, Nakashima-Y, Kojima-Y, Nobuyoshi-M, Kita-T, Tanaka-M.
  • Journal Title: Arterioscler Thromb Vase Biol. 28 Pages: 1077-83
  • Peer Reviewed
• [Journal Article] Small dense LDL-cholesterol relative to LDL-cholesterol is a strong independent determinant of hypoadiponectinemia in metabolic syndrome. (2008)
  • Author(s): Satoh-N, Wada-H, Ono-K, Yamakage-H, Yamada-K, Nakano-T, Hattori-M, Shimatsu-A, Kuzuya-H, Hasegawa-K.
  • Journal Title: Circulation J 72 Pages: 932-9
  • Peer Reviewed
• [Journal Article] statins activate GATA-6 and induce differentiated vascular smooth muscle cells. (2008)
  • Author(s): Wada H, Abe M, Ono K, Morimoto T, Kawamura T, Takaya T, Satoh N, Fujita M, Kita T, Shimatsu A, Hasegawa K.
  • Journal Title: Circulation J 374 Pages: 731-6
  • Peer Reviewed
• [Journal Article] Morphological and biochemical characterization of basal and starvation-induced autophagy in isolated adult rat cardiomyocytes. (2008)
  • Author(s): Maruyama R, Goto K, Takemura G, Ono K, Nagao K, Horie T, Tsujimoto A, Kanamori H, Miyata S, Ushikoshi H, Nagashima K, Minatoguchi S, Fujiwara T, Fujiwara H.
  • Journal Title: Am J Physiol Heart Circ Physiol. 295
  • Peer Reviewed
• [Journal Article] Identification of p300-targeted acetylated residues in GATA-4 during hyper trophic responses in cardiac myocytes. (2008)
  • Author(s): Takaya T, Kawamura T, Morimoto T, Ono K, Kita T, Shimatsu A, Hasegawa K.
  • Journal Title: J Biol Chem. 283 Pages: 9828-35
  • Peer Reviewed
• [Journal Article] Mulberry leaf ameliorates the expression profile of adipocytokines by inhibiting oxidative stress in white adipose tissue in db/db mice. (2008)
  • Author(s): Sugimoto M, Arai H, Tamura Y, Murayama T, Khaengkhan P, Nishio T, Ono K, Ariyasu H, Akamizu T, Ueda Y, Kita T, Harada S, Kamei K, Yokode M.
  • Journal Title: Atherosclerosis. (Epub ahead of print)
  • Peer Reviewed
• [Journal Article] Inhibition of CCR2 Aieliorates Insulin Resistance and Hepatic Steatosis in db/db Mice. (2008)
  • Author(s): Tamura Y, Sugimoto M, Murayama T, Ueda Y, Kanamori H, Ono K, Ariyasu H, Akamizu T, Kita T, Yokode M, Arai H.
  • Journal Title: Arterioscler Thromb Vase Biol. 28 Pages: 2195-201
  • Peer Reviewed
• [Presentation] Neural cell adhesion molecule is upregulated by metabolic stress and enhances the cardiomyocyte survival rate through neurite-cardiomyocyte communication. (2008)
  • Author(s): Nagao K, Ono K, Horie T, Koj ima Y, Nishi H, Kinoshita M, Kita T
  • Organizer: American Heart Association Annual Scientific Sessions
  • Place of Presentation: New Orleans, Louisiana
• [Presentation] NF-kB-dependent induction of microRNA-146 downregulates ErbB4 and may be involved in the deterioration of cardiomyocyte survival. (2008)
  • Author(s): Horie T, Ono K, Iwanaga Y, Nishi H, Nagao K, Kinoshita M, Mori R, Hasegawa K, Kita T
  • Organizer: American Heart Association Annual Scientific Sessions
  • Place of Presentation: New Orleans, Louisiana
• [Presentation] Upregulation of microRNA-27a, which downregulates retionoid X receptor a, controls cardiac myosin heavy chain gene expression in neonatal rat cardioyocyte (2008)
  • Author(s): Nishi H, Ono K, Kinoshita M, Nagao K, Horie T, Takanabe R, Hasegawa K, Iwanaga Y, Kita T
  • Organizer: American Heart Association Annual Scient fic Sessions
  • Place of Presentation: New Orleans, Louisiana
• [Presentation] Up-regulated expression of microRNA-143 in assocition with obesity in the adipose tissue of mice fed with high fat diet. (2008)
  • Author(s): Takanab R, Ono, Takaya T, Horie, T, Wada H, Kita T, Shimatsu A, Hasegawa K
  • Organizer: American Heart Association Annual Scienific Sessions
  • Place of Presentation: New Orlans, Louisiana
• [Presentation] IGF axis and adiponectin as prognostic markers of patients with congestive heart failure (2008)
  • Author(s): Watanabe S, Tamura T, Ono K, Kita, T, Kimura T, Furukawa Y
  • Organizer: European Society of Cardiology Congress
  • Place of Presentation: Munich, Germany
• [Presentation] miR-133は心筋細胞における代謝調節においてKLF15をターゲットとしてGLUT4の発現を制御している. (2008)
  • Author(s): 堀江貴裕, 尾野亘, 西仁勇, 長央和也, 木下美菜子, 森利依子, 岩永善高, 長谷川浩二, 北徹
  • Organizer: 第45回日本臨床分子医学会学術集会
  • Place of Presentation: 神戸
• [Presentation] miR-27aはラツト心筋細胞においてミオシン重鎖の発現を制御する (2008)
  • Author(s): 西仁勇, 尾野亘, 木下美菜子, 長央和也, 堀江貴裕, 岩永善高, 森利依子, 長谷川浩二, 北徹
  • Organizer: 第45回日本臨床分子医学会学術集会
  • Place of Presentation: 神戸
• [Presentation] microRNA-1の過剰発現はマウスES細胞の心筋分化を抑制する (2008)
  • Author(s): 高谷智英, 川村晃久, 尾野亘, 高鍋利依子, 鶏内伸二, 馬場志郎, 平家俊男, 中畑龍俊, 森本達也, 北徹, 島津章, 長谷川浩二
  • Organizer: 第45回日本臨床分子医学会学術集会
  • Place of Presentation: 神戸
• [Presentation] Trapping genes that encode secreted and transmembrane proteins in H9C2 cardiomyoblasts. (2008)
  • Author(s): Nagao K, Ono K, Horie T, Nishi H, Kinoshita M, Hasegawa K, Kita T
  • Organizer: 第31回心筋代謝研究会
  • Place of Presentation: 東京