ケトン体代謝異常症の分子病態とその原因酵素遺伝子の発現調節機構の解明

2009 Fiscal Year Annual Research Report

• Project/Area Number: 21591317
• Research Institution: Gifu University
• Principal Investigator: 深尾 敏幸 Gifu University, 大学院・医学系研究科, 教授 (70260578)
• Keywords: ケトン / 先天代謝異常症 / β-ケトチオラーゼ欠損症 / サクシニル-CoA:3-ケト酸CoAトランスフェラーゼ欠損症 / ケトアシドーシス / 遺伝子発現

Research Abstract

本年度もT2,SCOT欠損症の酵素診断,遺伝子診断は研究の一環として世界各地から受け入れ,酵素診断,遺伝子診断を行った.その1つはベトナムの8症例におけるコモン変異の同定にもつながった(Mol Genet Metab 2010).
ミスセンス変異の詳細な解析
1.T2欠損症のエクソン10内のサイレンス変異によるエクソンスキップの症例の解析では,エクソン10においてはSF2/ASFというexonic splicing enhancer配列が正常なスプライシングには必要で,その部位に変異が入ったためにスプライシング異常が生じていることを,患者変異を導入したmini-gene constructの作成するとともに,同じESE配列を減弱させる1塩基置換やauthentic splice部位を増強する1塩基置換を導入したmini-gene constructも作成し,transfectionによるスプライシング実験を行って明らかにし,論文はMol Genet Metabに受理された.
2.SCOT変異cDNAの発現実験を行い,同定された変異が病因となる変異であることを確認するとともに,一部は発現温度を30度,37度,40度で行うことで,温度感受性変異であることを明らかにした(現在投稿準備中)
MLPA法の確立
遺伝子内のエクソン領域の欠失を検出するために,SCOT遺伝子のMLPA法について検討を行っている.
SCOT遺伝子の肝臓特異的発現抑制機構の解明
1.SCOT遺伝子のプロモーター領域のCpG islandsがヒトでもマウスでも発現のみられない肝細胞においてもメチル化されていないことを明らかにした(Mol Med Reports 2010)
2.HepG2細胞(肝細胞特徴維持)においてSCOT遺伝子の転写がオンなのか,オフなのかについてChIP assayを用いて検討をしており,さらに解析をすすめている.

Research Products

• [Journal Article] Clinical Presentation in Siblings with Mitochondrial Acetoacetyl-CoA Thiolase Deficiency and Identification of Two Novel Mutations. (2010)
  • Author(s): Thummler S, Dupont D, Acquaviva C, Fukao T, de Ricaud D
  • Journal Title: Tohoku J Exp Med 220 Pages: 27-31
  • Peer Reviewed
• [Journal Article] CpG islands around exon 1 in Succinyl-CoA : 3-ketoacid CoA transferase (SCOT) gene were hypomethylated even in human and mouse hepatic tissues where SCOT gene expression was completely suppressed. (2010)
  • Author(s): Fukao T, Zhang G, Matsuo N, Kondo N
  • Journal Title: Molecular Medicine Reports 3 Pages: 335-359
  • Peer Reviewed
• [Journal Article] Decreased levels of metabolic enzymes in pancreatic islets of patients with type 2 diabetes. (2009)
  • Author(s): Macdonald MJ, Longacre MJ, Langberg EC, Tibell A, Kendrick MA, Fukao T, Ostenson CG.
  • Journal Title: Diabetologia 52 Pages: 1087-1091
  • Peer Reviewed
• [Journal Article] A novel molecular aspect of Japanese patients with medium-chain acyl-CoA dehydrogenase deficiency (MCADD) : c.449-452delCTGA is a common mutation in Japanese patients with MCADD. (2009)
  • Author(s): Purevsuren J, Kobayashi H, Hasegawa Y, Mushimoto Y, Li H, Fukuda S, Shigematsu Y, Fukao T, Yamaguchi S
  • Journal Title: Mol Genet Metab 96 Pages: 77-79
  • Peer Reviewed
• [Journal Article] Clinical and molecular investigations of 5 Japanese patients with mitochondrial trifunctional protein deficiency. (2009)
  • Author(s): Purevsuren J, Fukao T, Hasegawa Y, Fukuda S, Kobayashi H, Yamaguchi S.
  • Journal Title: Mol Genet Metab 98 Pages: 372-377
  • Peer Reviewed
• [Journal Article] (2009)
  • Author(s): 浦澤林太郎, 久保徹夫, 深尾敏幸
  • Journal Title: 症例から学ぶ先天代謝異常症～日常診療からのアプローチ～(診断と治療社) Pages: 100-102
• [Journal Article] A common mutation, R208X, identified in Vietnamese patients with mitochondrial acetoacetyl-CoA thiolase (T2) deficiency
  • Author(s): Fukao T, Nguyen HT, Nguyen NT, Vu DC, Can NTB, Pham ATV, Nguyen KN, Kobayashi H, Hasegawa Y, Bui TH, Niezen-Koning KE, Wanders RJA, de Koning T, Nguyen LT, Yamaguchi S, Kondo N
  • Journal Title: Mol Genet Metab (In press)
  • Peer Reviewed
• [Journal Article] A novel mutation (c.951C>T) in an exonic splicing enhancer results in exon 10 skipping in the human mitochondrial acetoacetyl-CoA thiolase gene
  • Author(s): 169. Fukao T, Horikawa R, Naiki Y, Tanaka T, Takayanagi M, Yamaguchi S, Kondo N
  • Journal Title: Mol Genet Metab (In press)
  • Peer Reviewed
• [Presentation] Clinical and laboratory features of mitochondrial acetoacetyl-CoA thiolase (T2) deficiency in Viet Nam (2010)
  • Author(s): Nguyen Thi Hoan, Nguyen Ngoc Khanh, Vu Chi Dung, Bui Phuong Thao, Can Thi Bich Ngoc, Khu Khanh Dung, Seiji Yamaguchi, Toshiyuki Fukao, Nguyen Thanh Liem.\
  • Organizer: The 1st Asian Congress for Inhetited Metabolic Diseases.
  • Place of Presentation: Fukuoka
  • Year and Date: 20100307-20100310
• [Presentation] A common mutation R208X identified in Vietnamese patients with mitochondrial acetoacetyl-CoA thiolase (T2) deficiency. (2010)
  • Author(s): Toshiyuki Fukao, Hoan Thi Nguyen, Nhan Thu Nguyen, Ngoc Thi Bich Can, Dung ChiVu, Anh Thi Van Pham, Khanh Ngoc Nguyen, Hironori Kobayashi, Yuki Hasegawa, Thao Phuong Bui, Liem Thanh Nguyen, Seiji Yamaguchi, Naomi Kondo.
  • Organizer: The 1st Asian Congress for Inhetited Metabolic Diseases.
  • Place of Presentation: Fukuoka
  • Year and Date: 20100307-20100310
• [Presentation] β-ケトチオラーゼ欠損症の分子病態:Exonic splicing enhancer変異の解析. (2009)
  • Author(s): 深尾敏幸, 堀川玲子, 内木康博, 田中藤樹, 近藤直実
  • Organizer: 第51回日本先天代謝異常学会総会
  • Place of Presentation: 東京
  • Year and Date: 20091105-20091107
• [Presentation] 同時に蓍しいケトアシドーシス発作を来した3歳の双生児例(T2遺伝子H144Pホモ接合体例). (2009)
  • Author(s): 丸山慎介, 豊島光雄, 鍬田直美, 深尾敏幸, 堀川玲子, 内木康博, 田中藤樹, 近藤直実,
  • Organizer: 第51回日本先天代謝異常学会総会
  • Place of Presentation: 東京
  • Year and Date: 20091105-20091107
• [Presentation] An exonic splicing mutation identified in a beta-ketothiolase-deficient patient. (2009)
  • Author(s): Fukao T, Naiki Y, Tanaka, Kondo N.
  • Organizer: 11th International Congress of Inborn Errors of Metabolism
  • Place of Presentation: SanDiego, USA
  • Year and Date: 20090829-20090902
• [Presentation] The Time-course of blood and urinary acylcarnitine levels during initial L-carnitine supplementation to a CPT2-deficient patient. (2009)
  • Author(s): Fukao T, Hori T, Teramoto T, Kobayashi H, Takayanagi M, Hasegawa Y, Yasuno T, Yamaguchi S, Kondo N.
  • Organizer: 11th International Congress of Inborn Errors of Metabolism
  • Place of Presentation: SanDiego, USA
  • Year and Date: 20090829-20090902
• [Presentation] Clinical and molecular investigations of 5 Japanese patients with mitochondrial trifunctional protein deficiency. (2009)
  • Author(s): Purevsuren J, Fukao T, Hasegawa Y, Kobayashi H, Fukuda S, Yamaguchi S.
  • Organizer: 11th International Congress of Inborn Errors of Metabolism
  • Place of Presentation: SanDiego, USA
  • Year and Date: 20090829-20090902
• [Presentation] エクソン内1塩基置換により,Exonic splicing Enhancer配列に影響し, エクソンスキップを来したミトコンドリアアセトアセチル-CoAチオラーゼ(T2)の1症例. (2009)
  • Author(s): 深尾敏幸, 近藤直実, 内木康博, 堀川玲子, 田中藤樹.
  • Organizer: 第54回日本人類遺伝学会
  • Place of Presentation: 東京
  • Year and Date: 20090824-20090826