Identification of target genes for DNA methylation in skeletal muscle and its medical application

骨骼肌DNA甲基化靶基因的鉴定及其医学应用

• 批准号: 23659468
• 负责人: OGAWA Yoshihiro
• 金额: $ 2.5万
• 依托单位: Tokyo Medical and Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Challenging Exploratory Research
• 财政年份: 2011
• 资助国家: 日本
• 起止时间: 2011 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-23659468/
• 关键词: 骨格筋; DNA; メチル化; エピジェネティクス; 生活習慣病; 遺伝子発現; DNAメチル化

DNA methylation is essential for normal embryonic development, and altered DNA methylation patterns have been implicated in tumorigenesis. An epigenetic mechanism involving DNA methylation has also been suggested to be involved in the regulation of metabolic processes ; however, the molecular basis of this mechanism has not been clearly demonstrated. In this study, we attempted to get an insight into the role of DNA methylation in skeletal muscle, which plays important roles in exercise, energy expenditure and glucose metabolism. We made skeletal muscle-specific knockout(KO) mice with Dnmt3a(a de novo DNA methyltransferase highly expressed in skeletal muscle) by crossing Dnmt3a flox/flox mice and transgenic mice expressing Cre recombinase, driven by the skeletal muscle alpha-actin promoter. A quantitative real-time PCR analysis confirmed that Dnmt3a mRNA levels were markedly diminished in the skeletal muscle but not in other tissues of the KO mice. In this study, a genome-wide DNA methylation analysis called Microarray-based Integrated Analysis of Methylation by Isoschizomers(MIAMI) was preformed using the methylation-sensitive restriction enzyme HpaII and a genome microarray. MIAMI analysis revealed a marked decrease in DNA methylation in the KO mice, including that of several genes coding transcription factors. Despite this, the decreased DNA methylation did not correlate with the gene expression levels under the same condition. Because DNA methylation is considered a key epigenetic contributor in the maintenance of gene silencing, gene expression in the KO mice may be modified in the presence of additional metabolic stress. Therefore, analysis of metabolic phenotype of the KO mice will be essential to elucidate the epigenetic regulation of skeletal muscle gene expression and skeletal muscle-related metabolic diseases.

DNA甲基化对正常胚胎发育至关重要，并且改变的DNA甲基化模式与肿瘤发生有关。涉及DNA甲基化的表观遗传机制也被认为参与代谢过程的调节;然而，这种机制的分子基础尚未得到明确证明。在这项研究中，我们试图了解骨骼肌中DNA甲基化的作用，骨骼肌在运动，能量消耗和葡萄糖代谢中起着重要作用。我们通过将Dnmt 3a flox/flox小鼠和表达Cre重组酶的转基因小鼠杂交，用Dnmt 3a（一种在骨骼肌中高度表达的从头DNA甲基转移酶）制造骨骼肌特异性敲除（KO）小鼠，由骨骼肌α-肌动蛋白启动子驱动。定量实时PCR分析证实，Dnmt 3a mRNA水平在骨骼肌中显著降低，但在KO小鼠的其他组织中没有。在这项研究中，一个全基因组的DNA甲基化分析称为基于微阵列的集成分析甲基化的异构体（迈阿密）进行甲基化敏感的限制性内切酶HpaII和基因组微阵列。迈阿密分析显示KO小鼠的DNA甲基化显著降低，包括编码转录因子的几个基因。尽管如此，在相同条件下，DNA甲基化的降低与基因表达水平无关。由于DNA甲基化被认为是维持基因沉默的关键表观遗传贡献者，因此在存在额外代谢应激的情况下，KO小鼠中的基因表达可能会发生改变。因此，分析KO小鼠的代谢表型对于阐明骨骼肌基因表达的表观遗传调控和骨骼肌相关代谢疾病是必不可少的。

项目成果

Epigenetic Modifications Underlying Insulin Resistance

胰岛素抵抗背后的表观遗传修饰

• 发表时间: 2011
• 作者: 大津義晃;中川祐子;小島至;窪田直人;小川佳宏;早崎 貴洋;吉良潤一;窪田直人;Yoshihiro Ogawa
• 通讯作者: Yoshihiro Ogawa

Melanocortin 4 Receptor-Deficient Mice as a Novel Mouse Model of Nonalcoholic Steatohepatitis

• DOI: 10.1016/j.ajpath.2011.07.014
• 发表时间: 2011-11-01
• 期刊: AMERICAN JOURNAL OF PATHOLOGY
• 影响因子: 6
• 作者: Itoh, Michiko;Suganami, Takayoshi;Ogawa, Yoshihiro
• 通讯作者: Ogawa, Yoshihiro

エピジェネティスク機構による細胞制御と病態

表观遗传机制的细胞调控和病理学

• 发表时间: 2011
• 作者: Kataoka K;et al;杉山 斉;吉良潤一ら;大津 義晃;小川佳宏;吉良潤一ら;平松 英樹;Kojima I;吉良潤一;窪田直人,窪田哲也,門脇孝;伊藤 功;小川佳宏
• 通讯作者: 小川佳宏

生活習慣病のエピゲノム制御

生活方式相关疾病的表观基因组控制

• 发表时间: 2011
• 作者: 増淵洋祐;中川祐子;馬金輝;小島至;中島 諒子;岩部美紀,山内敏正,岩部真人,窪田直人,門脇孝;中川 祐子;Yoshihiro Ogawa;吉良潤一ら;Yoshihiro Ogawa;窪田哲也,窪田直人,佐藤寛之,岩村智勝,井上真理子,林高則,山内敏正,植木浩二郎,門脇孝;伊藤 恭彦;吉良潤一ら;大津 義晃;桜井賛孝,高本偉碩,窪田直人,熊谷勝義,小畑淳史,植木浩二郎,門脇孝;小川佳宏
• 通讯作者: 小川佳宏

栄養とエピジェネティクス

营养和表观遗传学

• 作者: 小島至;中川祐子;吉良潤一ら;小川佳宏
• 通讯作者: 小川佳宏