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 名发生急性肝衰竭的无关日本红细胞生成性原卟啉症 (EPP) 患者中是否存在亚铁螯合酶 (FECH) 基因突变，从外周血白细胞中提取基因组 DNA，通过扩增 FECH 基因的每个外显子并进行单链检查。构象多态性（SSCP）分析，以及随后的直接测序来寻找突变，也被应用于谱系分析。在 5 名不相关的日本患者中总共鉴定出 4 个独特的突变，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。特定的限制性内切酶或显示特定的 SSCP 模式，并且可以通过特定酶或特定的 SSCP 模式通过基因组 DNA 的扩增片段进行筛选。这项研究在以前报道的突变的基础上添加了一些新的突变，同时还添加了一些新的突变。然而，我们无法确切地阐明肝脏并发症的原因，因为他们的家族中有无症状的携带者，他们表现出相同的突变，但从未发展为肝脏疾病，因此我们现在正在尝试分析其他正常等位基因。称为め低表达も野生型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。

Yutaka Hone, Takahiko Matsuura, Yoshiaki Santo, Kazuki Godai, Masako Adachi, Naoto Maeda, et al.: "A one year and 3 months experience of couselling porphyrias in our hospital"Jpn Pharmacoi Ther. 31. S73-S75 (2003)

Yutaka Hone、Takahiko Matsuura、Yoshiaki Santo、Kazuki Godai、Masako Adachi、Naoto Maeda 等：“我院一年零 3 个月的卟啉症辅导经验”Jpn Pharmacoi Ther.