Study on biochemical and molecular evoluation for mitochondrial β-oxidation defects

线粒体β-氧化缺陷的生化和分子进化研究

基本信息

批准号：
10470178
负责人：
YAMAGUCHI Seiji
金额：
$ 3.2万
依托单位：
Shimane Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B).
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2000
项目状态：
已结题

项目摘要

Mitochondrial fatty acid beta-oxidation disorders (FAOD) has atracted attention recently. We investigated the acutual situation of Japanese patients with FAOD, and studied the biochemical evaluation and molecular analysis of FAOD patients.1) Survey of Japanese patients with FAOD : We found 62 Japanese patients with FAOD during the period between 1985 to 2000, using questionnaires, literatures and personal communications. The number of patients was largest in CPT2 deficiency (14), second was glutaric acidemia type 2 (12). The patients in early childhood tend to have acute encephalopathy-like illness, whereas patients in older childhood or adolescence did myopathic illness.2) Establishment of a simplified GC/MS data processing system : FAOD patients often show nonkletotic dicarboxylic aciduria in organic acidemia screening using GC/MS.We established a personal computer-based system of automated metabolic profiling and diagnosis.3) Measurement of free fatty acids in dried blood filter paper : For the screening of FAOD, acylcarnitine analysis by tandem MS has become popular. Tandem MS may potentially fail to detect such disorders as a false-negative. We established a simplified, sensitive method to measure the blood free fatty acid levels in blood filter paper.4) Detection of FAOD by incorporation of RI-labeled fatty acids using cultured cells : We established a screening system for FAOD by incorporation of RI-labeled fatty acids, including palmitate and myristate, analyzing cells from VLCAD deficiency, GA2 and CPT2 deficiency.5) Molecular analysis of Japanese patients with VLCAD deficiency and GA2 : We identified genetic mutations in five VLCAD deficient patients and three GA2 patients. In VLCAD deficiency, the genotype/phenotype correlation was apparent and the temperature sesitivity was seen among mutations detected. In GA2, we identified the first Japanese case of ETF-alpha subunit deficiency in two patients by pulse labeling and DNA analysis.

线粒体脂肪酸β-氧化障碍（FAOD）最近引起了人们的关注。我们调查了日本FAOD患者的实际情况，并对FAOD患者的生化评估和分子分析进行了研究。1）日本FAOD患者的调查：我们在1985年至2000年期间通过问卷调查的方式发现了62名日本FAOD患者，文学和个人通讯。 CPT2缺乏症患者数量最多（14例），其次是2型戊二酸血症（12例）。幼儿期的患者往往患有急性脑病样疾病，而大龄儿童或青春期的患者则患有肌病。 2) 建立简化的 GC/MS 数据处理系统：FAOD 患者在有机酸血症筛查中经常表现出非收缩性二羧酸尿症GC/MS。我们建立了基于个人计算机的自动代谢分析和诊断系统。3) 干血滤纸中游离脂肪酸的测量：用于筛查FAOD，通过串联质谱进行酰基肉碱分析已变得流行。串联 MS 可能无法检测出此类疾病，导致假阴性。我们建立了一种简化、灵敏的方法来测量血液滤纸中的血液游离脂肪酸水平。4) 使用培养细胞掺入 RI 标记的脂肪酸来检测FAOD：我们建立了一种通过掺入 RI 标记的脂肪酸来检测FAOD 的系统脂肪酸，包括棕榈酸和肉豆蔻酸，分析 VLCAD 缺陷、GA2 和 CPT2 缺陷的细胞。5) 对日本 VLCAD 缺陷和 GA2 患者的分子分析：我们发现了基因突变5 名 VLCAD 缺陷患者和 3 名 GA2 患者。在 VLCAD 缺陷中，基因型/表型相关性很明显，并且在检测到的突变中发现了温度敏感性。在 GA2 中，我们通过脉冲标记和 DNA 分析在两名患者中发现了日本首例 ETF-α 亚基缺陷病例。