先天性ケトン体代謝異常症の分子病態-サクシニル-CoA:3-ケト酸CoAトランスフェラーゼ欠損症を中心として-

• 研究課題/領域番号: 11670754
• 研究種目: 基盤研究(C)
• 配分区分: 補助金
• 応募区分: 一般
• 研究分野: 小児科学
• 研究機関: 岐阜大学
• 研究代表者: 深尾 敏幸 岐阜大学, 医学部・附属病院, 助手 (70260578)
• 研究分担者: 伊上 良輔 岐阜大学, 医学部・附属病院, 助手 (60273132) ; 祐川 和子 岐阜大学, 医学部, 助手 (60115409)
• 研究期間 (年度): 1999 – 2000
• 研究課題ステータス: 完了 (2000年度)
• 配分額: 1,500千円 (直接経費: 1,500千円); 2000年度: 1,500千円 (直接経費: 1,500千円)
• キーワード: β-ケトチオラーゼ欠損症 / ミトコンドリア・アセトアセチル-CoAチオラゼ / サクシニル-CoA:3-ケト酸CoAトランスフェラーゼ / 遺伝子クローニング / 遺伝子変異 / 発現調節 / ケトン体 / ミトコンドリア・アセトアセチル-CoAチオラーゼ / T2欠損症 / サクシニル-CoA:3ケト酸CoAトランスフェラーゼ / サクシニル-CoA:3ケト酸CoAトランスフェラーゼ欠損症 / ミトコンドリア.アセトアセチル-CoAテオラーゼ

研究概要

SCOT遺伝子の構造,配列を明らかにし,ヒトSCOT蛋白の3次構造モデルを作成し,3人のSCOT欠損症患者の遺伝子変異を同定,その特徴を明らかにした.ゲノム遺伝子の配列をあきらかにしたことで,SCOT欠損症患者遺伝子変異のゲノムDNAでのスクリーニングを可能とした.またSCOT遺伝子がなぜ正常肝細胞で発現しないのかをあきらかにするためSCOT上流域の解析を行った.約3kbの配列を決定し,基本的なプロモーターにはSp1が結合して活性化されることを明らかにし,肝臓特異的な抑制に関与すると思われる領域を検討した。またT2欠損症については,スペイン5症例の遺伝子変異について変異の特徴を3次構造モデルを用いて検討した.また一例にみられた380C>Tはエクソン5上にあり,この変異によってエクソン内のすでに存在するcryptic splice部位が活性化され,ほとんどがこの部位でスプライシングが生じることを明らかにした.またこれまでに遺伝子変異を決定した26例について主治医にアンケート調査を行い,臨床経過,予後,遺伝子型と臨床型の関連について調べた.この報告はT2欠損症ではじめて行われた臨床追跡調査であり,意義深い.T2欠損症は診断さえつけば,以後の重篤な発作は軽度の蛋白制限などにより防がれており,予後もこれまで報告されていた以上によいことがわかった.また臨床的重症度は遺伝子型と明らかな相関はなかった。これまでの成果が評価され,Methods in Enzymology,Metabolic ＆ Molecular Bases of Inherited Disease(8th edition)の章を共同執筆した.

研究成果

• [文献書誌] Fukao T: "Succinyl-CoA : 3-ketoacid CoA transferase (SCOT) : Cloning of human SCOT gene, tertiary structural modeling of the human SCOT monomer, and Characterization of three pathogenic mutations."Genomics. 68. 144-151 (2000)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Watanabe H: "Molecular basis of very-long-chain acyl-CoA dehydrogenase deficiency in 3 lsraeli patients : identification of a complex mutant allele with P65L and K247Q mutations, the former being an exonic mutation causing exon 3 skipping."Human Mutation. 15. 430-438 (2000)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Doi T: "Milder childhood form of very long-chain acyl-CoA dehydrogenase deficiency in a 6-year-old Japanese boy."European Journal of Pediatrics. 159. 908-911 (2000)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Fukao T: "Myopathic form of very-long chain acyl-CoA dehydrogenase deficiency : Evidence for temperature-sensitive mild mutations in both mutant alleles in a Japanese girl."Pediatric Research. (in press).
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Fukao T: "The clinical phenotype and outcome of mitochondrial acetoacetyl-CoA thiolase deficiency (Beta-ketothiolase or T2 deficiency) in 26 enzymatically proved and mutation-defined patients."Molecular Genetics and Metabolism. (in press).
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Gibson KM: "Molecular and enzymatic methods for detection of genetic defects in the distal pathways of branched-chain amino acid metabolism.in Branched chain amino acids, part B"Methods in Enzymology. 324. 432-453 (2000)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Nakamura K: "A novel single base substitution (380C>T) that activates a 5 bases-downstream cryptic splice-acceptor site within exon 5 in almost all transcripts in the human mitochondrial acetoacetyl-CoA thiolase gene."Molecular Genetics and Metabolism. (in press).
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Mitchell GA: "Chapter 102 Inborn errors of ketone body catabolism.In Metabolic and Molecular Bases of Inherited Disease (8th edition)"McGraw-Hill. 30 (2001)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Fukao T, Mitchell GA, Song X-Q, Nakamura H, Kassovska-Bratinova S, Orii KE, Wraith JE, Besley G, Wanders RJA, Niezen-Koning KE, Berry GT, Palmieri M, Kondo N.: "Succinyl-CoA : 3-ketoacid CoA transferase (SCOT) : Cloning of human SCOT gene, tertiary structural modeling of the human SCOT monomer, and characterization of three pathogenic mutations."Genomics. 68. 144-151 (2000)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Watanabe H, Orii EK, Fukao T, Song X-Q, Aoyama T, IJIst L, Ruiter J, Wanders RJA, Kondo N.: "Molecular basis of very-long-chain acyl-CoA dehydrogenase deficiency in 3 Israeli patients : identification of a complex mutant allele with P65L and K247Q mutations, the former being an exonic mutation causing exon 3 skipping."Hum Mutat. 15. 430-438 (2000)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Doi T, Abo W, Tateno M, Hayashi K, Hori T, Nakada T, Fukao T, Takahashi Y, Terada N.: "Milder childhood form of very long -chain acyl-CoA dehydrogenase deficiency in a 6-year-old Japanese boy."Eur J Pediatr. 159. 908-911 (2000)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Fukao T, Watanabe H, Orii KE, Takahashi Y, Hirano A, Kondo T, Yamaguchi S, Aoyama T, Kondo N.: "Myopathic form of very-long chain acyl-CoA dehydrogenase deficiency : Evidence for temperature-sensitive mild mutations in both mutant alleles in a Japanese girl."Pediatr Res. (in press).
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Fukao T, Scriver CR, Kondo N and T2 Collaborative Working Group.: "The clinical phenotype and outcome of mitochondrial acetoacetyl-CoA thiolase deficiency (Beta-ketothiolase or T2 deficiency) in 26 enzymatically proved and mutation-defined patients."Mol Genet Metab. (in press).
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Gibson K, Ugarte M, Fukao T, Mitchell GA.: "Molecular and enzymatic methods for detection of genetic defects in the distal pathways of branched-chain amino acid metabolism. in Branched chain amino acids, part B"Methods in Enzymology. 324. 432-453 (2000)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Nakamura K, Fukao T, Perez-Cerda C, Luque C, Song X-Q, Naiki Y, Kohno Y, Ugarte M, Kondo N.: "A novel single base substitution (380C>T) that activates a 5 bases-downstream cryptic splice-acceptor site within exon 5 in almost all transcripts in the human mitochondrial acetoacetyl-CoA thiolase gene."Mol Genet Metab. (in press).
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Mitchell GA, Fukao T.: "Chapter 102 Inborn errors of ketone body catabolism. Metabolic and Molecular Bases of Inherited Disease (8th edition)"McGraw-Hill, Inc. 2327-2356 (30) (2000)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Fukao T: "Succinyl-CoA : 3-ketoacid CoA transferase (SCOT) : Cloning of human SCOT gene, tertiary structural modeling of the human SCOT monomer, and characterization of three pathogenic mutations."Genomics. 68. 144-151 (2000)
• [文献書誌] Watanabe H: "Molecular basis of very-long-chain acyl-CoA dehydrogenase deficiency in 3 Israeli patients : identification of a complex mutant allele with P65L and K247Q mutations, the former being an exonic mutation causing exon 3 skipping."Human Mutation. 15. 430-438 (2000)
• [文献書誌] Doi T: "Milder childhood form of very long-chain acyl-CoA dehydrogenase deficiency in a 6-year-old Japanese boy."European Journal of Pediatrics. 159. 908-911 (2000)
• [文献書誌] Fukao T: "Myopathic form of very-long chain acyl-CoA dehydrogenase deficiency : Evidence for temperature-sensitive mild mutations in both mutant alleles in a Japanese girl."Pediatric Research. (in press).
• [文献書誌] Fukao T: "The clinical phenotype and outcome of mitochondrial acetoacetyl-CoA thiolase deficiency (Beta-ketothiolase or T2 deficiency) in 26 enzymatically proved and mutation-defined patients."Molecular Genetics and Metabolism. (in press).
• [文献書誌] Gibson KM: "Molecular and enzymatic methods for detection of genetic defects in the distal pathways of branched-chain amino acid metabolism. in Branched chain amino acids, part B"Methods in Enzymology. 324. 432-453 (2000)
• [文献書誌] Nakamura K: "A novel single base substitution (380C>T) that activates a 5 bases-downstream cryptic splice-acceptor site within exon 5 in almost all transcripts in the human mitochondcrial acetoacetyl-CoA thiolase gene."Molecular Genetics and Metabolism. (in press).
• [文献書誌] Mitchell GA: "Chapter 102 Inborn errors of ketone body catabolism.In Metabolic and Molecular Bases of Inherited Disease (8th edition)"McGraw-Hill. 30 (2001)
• [文献書誌] Fukao T.: "ATM is upregulated during the mitogenic Response in peripheral blood mononuclear cells"Blood. 94・6. 1998-2006 (1999)
• [文献書誌] Fukao T.: "Immunoblot analysis for laboratory diagnosis of Ataxia-telangrectasia:use of Epstein-Barrvirus transformed or phytohemagglutinin-stimlated lympholasts"Journal of Investigational Allergology and Clinical Immunology. 10(in press). (2000)
• [文献書誌] Watanabe H.: "Molecular Bases of very-long-chain acyl-CoA dehydrogenase deficiency:Identification of a complex mutant allele with P65L and K247Q mutations,theformer being an exonic mutation causing exon 3 skipping"Human Mutation. (in press). (2000)
• [文献書誌] Kaneko H.: "Expression of the BLM gene in human haematopoietic cells"Clin.Exp.Immunol. 118. 285-289 (1999)
• [文献書誌] Inoue R.: "Time-course study of the levels of urimary leukotriene E4,serum thromboxane B2 and serum eosingihil cationic protein in spentaneous asthima attacks in fire children."Journal of Investigational allergology and Clinical Immunology. 9・6. 361-366 (1999)
• [文献書誌] 浦澤林太郎: "急激な意識障害で発症したβ-ケトチオラーゼ欠損症の1乳児例"日本小児科学会雑誌. 103・8. 849-852 (1999)
• [文献書誌] Gibson K.: "Molecular and enzymatic methods for detection of genetic defects in the distal pathways of branched-chain amino acid metabolism In Branched Chain amino acids,part B"Academic Press(in press).
• [文献書誌] Mitchell GA: "Inborn errors of Ketone body catabolism. In Molecular and Metabolic Bases of Inherited Disease"McGraw-HIll Inc(in press).