Molecular basis of inborn errors of ketone body metabolism, especially succinyl-CoA : 3-ketoacid CoA transferase deficiency

酮体代谢先天性缺陷，尤其是琥珀酰辅酶A：3-酮酸辅酶A转移酶缺乏症的分子基础

• 批准号: 11670754
• 负责人: FUKAO Toshiyuki
• 金额: $ 0.96万
• 依托单位: Gifu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670754/
• 关键词: beta-ketothiolase deficiency; mitochondrial acetoacetyl-CoA thiolase; succinyl-CoA : 3-ketoacid CoA transferase; gene cloning; gene mutation; gene regulation; ketone body; 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

1) SCOT deficieicny : We revealed the structure and sequence of human SCOT gene, made tertiary structural model of human SCOT protein, and identified and characterized gene mutations in 3 SCOT deficient patients. We now can screen SCOT gene mutations at the genomic level. We analyzed 5' flanking regions of human SCOT gene in order to clarify the mechanism for specific SCOT gene supression in hepatocytes. We determined the sequence of 3 kb in the region, and revealed that Sp1 basically drive the SCOT gene expression. We also searched cis-elements responsible for specific supression in hepatocytes.2) T2 deficiency : We analyzed character of amino acid alternations (gene mutations) identified in 5 spanish T2 deficient patients on teritiary structural model of T2 protein. In one patient, we identified a novel splicing mutation 380C>T.This mutation located on exon 3 activated pre-exsisting cryptic splice site on exon 3, resulting in exclusive splicing at the cryptic site. We have identified gene mutations in 26 T2 deficient patients. We collected clinical information from the physicians for these patients with the use of questionnaire to search clinical courses and outcome and to clarifiy genotype/phenotype correlation in T2 deficiency. By the study, we concluded that (1) T2 deficiency has a favorable outcome in general ; (2) severe ketoacidotic attack can be avoidable after confirmation of the diagnosis ; (3) there is no apparent phenotype/genotype correlation.The head investigator was honored to write chapters in "Methods in Enzymology" and in "Metabolic ＆ Molecular Bases of Inherited Disease".

1) SCOT缺乏症：我们揭示了人SCOT基因的结构和序列，建立了人SCOT蛋白三级结构模型，并对3例SCOT缺乏症患者的基因突变进行了鉴定和表征。我们现在可以在基因组水平上筛选SCOT基因突变。我们分析了人SCOT基因的5'侧翼区域，以阐明肝细胞特异性SCOT基因抑制的机制。我们在该区域确定了3kb的序列，发现Sp1基本驱动SCOT基因的表达。我们还搜索了肝细胞中负责特异性抑制的顺式元件。2) T2缺乏：我们分析了5例西班牙T2缺乏患者在T2蛋白三级结构模型上发现的氨基酸改变（基因突变）特征。在一名患者中，我们发现了一种新的剪接突变380C >t。这个位于外显子3的突变激活了外显子3上预先存在的隐剪接位点，导致隐剪接位点的独占剪接。我们在26例T2缺陷患者中发现了基因突变。我们从医生那里收集了这些患者的临床信息，使用问卷调查来搜索临床过程和结果，并澄清T2缺乏的基因型/表型相关性。通过研究，我们得出结论：(1)总的来说，T2缺乏具有良好的结果；(2)经确诊，严重酮症酸中毒发作可避免；(3)表型/基因型相关性不明显。首席研究员荣幸地在“酶学方法”和“遗传疾病的代谢与分子基础”中撰写了章节。

项目成果

Mitchell GA: "Inborn errors of Ketone body catabolism. In Molecular and Metabolic Bases of Inherited Disease"McGraw-HIll Inc(in press).

米切尔 GA：“酮体分解代谢的先天性错误。遗传性疾病的分子和代谢基础”McGraw-HIll Inc（正在出版）。

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)

Doi T：“一名 6 岁日本男孩患有较轻的儿童期极长链酰基辅酶 A 脱氢酶缺乏症。”《欧洲儿科杂志》。

Fukao T.: "ATM is upregulated during the mitogenic Response in peripheral blood mononuclear cells"Blood. 94・6. 1998-2006 (1999)

Fukao T.：“ATM 在外周血单核细胞促有丝分裂反应期间上调”血液 94・6（1999）。

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)

Fukao T.：“共济失调毛细血管扩张症实验室诊断的免疫印迹分析：使用 Epstein-Barrvirus 转化或植物血凝素刺激的淋巴母细胞”研究变态反应学和临床免疫学杂志。

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).

Fukao T：“26 名经酶法证明和突变定义的患者中线粒体乙酰乙酰辅酶 A 硫解酶缺乏症（β-酮硫解酶或 T2 缺乏症）的临床表型和结果。”分子遗传学和代谢。