Research on the epigenetics and environmental factors in erythropoietlc protoporphyria

红细胞生成性原卟啉症的表观遗传学及环境因素研究

• 批准号: 14570470
• 负责人: MAEDA Naoto
• 金额: $ 1.28万
• 依托单位: Tottori University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570470/
• 关键词: ervthropoletic protoporphyria; gene analysis; epigenetics

We investigated the presence of mutations of ferrochelatase (FECH) gene in 5 unrelated Japanese erythropoietic protoporphyria (EPP) patients who developed acute hepatic failure. Genomic DNA extracted from peripheral blood leukocytes, was examined by amplifying each exon of the FECH gene and performing single strand conformation polymorphism (SSCP) analysis, followed by direct-sequencing to look for mutations. Molecular analysis, was also applied for the pedigree analyses.A total of 4 unique mutations were identified in 5 unrelated Japanese patients. Each mutation encodes truncated protein, and the activity of the FECH expressed in a E.coli expression system was decreased compared to the normal control. These mutations changed cleavage sites of the specific restriction enzyme or showed specific SSCP patterns, and could be screened by an amplified fragment from genomic DNA by the specific enzymes or specific SSCP patterns.This study adds some new mutations to those that have been previously reported together with a concurrent and additional cause of liver failure. However, we could not exactly elucidate the reason of liver complications, because there are asymptomatic carriers in their families who showed the same mutations but never develop hepatic disorder. We are now trying to analyze the other normal allele, so-called メ low expressed モ wild-type FECH allele.

我们调查了5例发生急性肝功能衰竭的无关日本红细胞生成性原卟啉症（EPP）患者的亚铁螯合酶（FECH）基因突变。从外周血白细胞中提取的基因组DNA通过扩增FECH基因的每个外显子并进行单链构象多态性（SSCP）分析进行检查，然后直接测序以寻找突变。分子生物学分析也被应用于家系分析，在5名无关的日本患者中共鉴定出4种独特的突变。每个突变编码截短的蛋白质，并且与正常对照相比，在大肠杆菌表达系统中表达的FECH的活性降低。这些突变改变了特异性限制性内切酶的酶切位点或表现出特异性的SSCP图谱，并可通过特异性酶切或特异性SSCP图谱从基因组DNA中扩增出的片段进行筛选，本研究在以往报道的突变基础上增加了一些新的突变，同时也增加了一个肝衰竭的病因。然而，我们不能确切地阐明肝脏并发症的原因，因为在他们的家庭中有无症状携带者显示相同的突变，但从未发生肝脏疾病。我们现在正试图分析另一个正常的等位基因，即所谓的低表达野生型FECH等位基因。

项目成果

Naoto Maeda, Masahiko Miura, Yoshikazu Murawaki.: "Molecular defects in the ferrochelatase gene in Japanese patients of erythropoietic protoporphyria with severe live complications"Jpn Pharmacoi Ther. 31. S67-S71 (2003)

Naoto Maeda、Masahiko Miura、Yoshikazu Murawaki.：“患有严重并发症的日本红细胞生成性原卟啉症患者中亚铁螯合酶基因的分子缺陷”Jpn Pharmacoi Ther。

Naoto Maed, Masahiko Miura, Akihide Hosoda, Yoshikazu Murawaki, Hironaka Kawasaki.: "Genetic analysis in erythropoietic protoporphyria patients with severe hepatic disorders"J.Gastroenterol hepatol.. 17 (Suppl 3). A61 (2002)

Naoto Maed、Masahiko Miura、Akihide Hosoda、Yoshikazu Murawaki、Hironaka Kawasaki.：“患有严重肝脏疾病的红细胞生成性原卟啉症患者的基因分析”J.Gastroenterol hepatol.. 17（增刊 3）。

前田直人, 三浦将彦, 村脇義和: "肝障害を合併した骨髄性プロトポルフィリン症におけるフェロケラターゼ遺伝子の解析"薬理と臨床. 37. S67-S71 (2003)

Naoto Maeda、Masahiko Miura、Yoshikazu Murawaki：“骨髓原卟啉症并发肝损伤的亚铁螯合酶基因分析”药理学和临床研究 37。S67-S71 (2003)。

Naoto Maeda, et al.: "Genetic analysis in erythropoietic protoporphyria patients with severe hepatic disorders"Journal of Gastroenterology and Hepatology. 17(Suppl). A61 (2002)

Naoto Maeda 等人：“患有严重肝脏疾病的红细胞生成性原卟啉症患者的基因分析”胃肠病学和肝脏病学杂志。

Naoto Maeda, et al.: "Molecular Defects in the Ferrochelatase Gene in Japanese Patients of Erythropoietic Protoporphyria with Severe Liver Complications"Gut. 51(Suppl III). A134 (2002)

Naoto Maeda 等人：“患有严重肝脏并发症的红细胞生成性原卟啉症日本患者中铁螯合酶基因的分子缺陷”Gut。