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Diagnoses and molecular bases of mitochondrial respiratory chain disorders

线粒体呼吸链疾病的诊断和分子基础

基本信息

批准号：
19591220
负责人：
OHTAKE Akira
金额：
$ 2.91万
依托单位：
Saitama Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2007
资助国家：
日本
起止时间：
2007 至 2009
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-19591220/
关键词：
遺伝・先天異常学ミトコンドリア病先天性高乳酸血症 Blue Native電気泳動皮膚線維芽細胞ミトコンドリア呼吸鎖異常症イムノブロット解析活性染色ミトコンドリアDNA枯渇症候群ミトコンドジアDNA枯渇症候群

项目摘要

BACKGROUND : Congenital and primary lactic acidosis is one of the most frequent inborn errors of metabolism, of whom only 30% have had its precise cause identified. Our aim is to make a prompt and correct diagnosis of mitochondrial respiratory chain disorders (MRCD), using Blue Native Polyacrylamide Gel Electrophoresis (BN-PAGE) combined with conventional enzyme assay. METHODS : Activities of the individual respiratory chain complexes and the mitochondrial matrix marker enzyme citrate synthase were measured in liver, heart and muscle homogenates and mitochondrial fractions isolated from cultured fibroblasts. Tissue homogenates or mitochondri isolated from skin fibroblasts were solubilised in n-dodecyl-maltoside and subjected to 4-13% BN-PAGE and western blotting using monoclonal antibodies specific for Complex I to IV subunits. Mitochondrial DNA and nuclear DNA copy numbers within tissues were determined by quantitative polymerase chain reaction. RESULTS : One hundred and ten patients were diagnosed to have MRCD out of 267 candidate patients. Most frequent was complex I deficiency, of whom many patients had tissue-specific type deficiency. Twenty patients out of 110 had mitochondrial DNA pathogenic mutations, which meant the majority of childhood-onset MRCD was nuclear origin. Patients with mtDNA mutations had milder symptoms than those suspected to have nuclear mutation. MtDNA depletion syndrome (MDS) was a prevalent cause of multiple MRCD. Twelve patients in 10 families were diagnosed to have hepatic MDS, and 5 patients were diagnosed to have myopathic MDS. Out of 12 hepatic MDS, we discovered nuclear genes mutations of DGUOK, POLG and MPV17 in 6 atients from 4 families. CONCLUSION : We must have a suspicion that almost every disease may be a MRCD. BN-PAGE is a useful guide to prompt and correct diagnosis, and future molecular analysis for categorizing respiratory chain disorders.

背景：先天性和原发性乳酸性酸中毒是最常见的先天性代谢错误之一，其中只有30%的人有明确的病因。我们的目的是利用Blue Native Polyacrylamide Gel Electrophoresis （BN-PAGE）结合常规酶分析法，对线粒体呼吸链疾病（MRCD）进行及时、正确的诊断。方法：测定肝脏、心脏和肌肉匀浆及培养成纤维细胞线粒体组分中单个呼吸链复合物和线粒体基质标记酶柠檬酸合成酶的活性。从皮肤成纤维细胞中分离的组织匀浆或线粒体溶解在正十二烷基麦糖苷中，用4-13%的BN-PAGE和针对复合物I至IV亚基的单克隆抗体进行western印迹。采用定量聚合酶链反应测定组织内线粒体DNA和细胞核DNA拷贝数。结果：267例候选患者中有110例被诊断为MRCD。最常见的是复合物I缺乏症，其中许多患者有组织特异性类型缺乏症。110名患者中有20名患有线粒体DNA致病性突变，这意味着大多数儿童发病的MRCD是核源性的。mtDNA突变患者的症状较疑似核突变患者轻。MtDNA缺失综合征（MDS）是多发MRCD的常见病因。10个家族中12例被诊断为肝性MDS， 5例被诊断为肌病性MDS。在12例肝MDS患者中，我们在4个家族的6例患者中发现DGUOK、POLG和MPV17核基因突变。结论：我们必须怀疑几乎每一种疾病都可能是MRCD。BN-PAGE是快速正确诊断和未来分子分析呼吸链疾病分类的有用指南。

项目成果

期刊论文数量（0）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

A case of Ehlers-Danlos syndrome type IV vascular type, demonstrated a newly recognized point mutation in the COL3A1 gene

埃勒斯-当洛斯综合征 IV 型血管型一例，显示 COL3A1 基因中新发现的点突变

DOI：
发表时间：
2010
期刊：
Inter Med 49(in press)
影响因子：
0
作者：
Komaki H;Nishigaki Y;Fuku N;Hosoya H;Murayama K;Ohtake A;Goto YI;Wakamoto H;Koga Y;Tanaka M;Komaki H;Sadakata R
通讯作者：
Sadakata R

Constitutively activated ALK-2 and increased Smadl/5 cooperatively induce BMP signaling in fibrodysplasia ossificans progressiva.

持续激活的 ALK-2 和增加的 Smadl/5 协同诱导进行性骨化性纤维发育不良中的 BMP 信号转导。