GRIM19 DNA甲基化介导的代谢改变导致口腔上皮衰老

Mitochondrial dysfunction and DNA methylation have been associated with aging. GRIM19 is a component of mitochondrial respiratory chain complex I. We found that GRIM19 DNA was partially methylated in healthy people and was associated with age. In this study, we will analyze the methylation status of the whole genome and precise sites in the promoter region of GRIM19 from oral epithelial tissues of different age groups of healthy subjects. We will utilize human normal oral keratinocyte (HNOK) with over-expression /knock-down GRIM19 and inducible GRIM19 knockout mouse model to analyze 1) changes of metabolic products, such as oxygen consumption, ROS, ATP, and NAD +; 2) the oxidation reduction sensors, such as mTOR, AMPK and Sirt1; 3) other markers of aging, such as p53, p16, HIF-1α, β- galactosidase and cloning ability. Thus, we will be able to reveal the effect of GRIM19 on mitochondrial metabolic function in the oral epithelium of healthy people, and to address the signal pathway of GRIM19 during aging. Meanwhile, through the HNOK aging model, we will observe changes of genome-wide methylation, GRIM19 methylation, and GRIM19 mRNA/protein in order to explore the effect of aging on DNA methylation in oral epithelial cells. The success of this study will be a major breakthrough in the understanding of the mechanism of oral epithelial aging and provide new information for the prevention of aging related diseases.

线粒体功能障碍和DNA甲基化与衰老相关。GRIM19是线粒体呼吸链复合物I组成成分。我们发现健康人口腔上皮细胞GRIM19部分甲基化，且与年龄有关。本项目将分析不同年龄健康人口腔上皮全基因组甲基化和GRIM19启动子甲基化精确位点和范围。将应用过表达/敲弱 GRIM19的人正常口腔上皮细胞（HNOK）和诱导性敲除GRIM19小鼠模型检测1)氧耗量、ROS、ATP、NAD+代谢物变化2)mTOR、AMPK、Sirt1氧化还原传感器3)p53、p16、HIF-1α、β-半乳糖苷酶和克隆能力衰老标志，揭示GRIM19对健康者口腔上皮线粒体代谢功能影响，阐述GRIM19衰老信号途径。同时，通过HNOK衰老模型观察全基因组和GRIM19甲基化、GRIM19mRNA/蛋白质的变化，探索衰老对口腔上皮甲基化的影响。本项目的完成对了解口腔上皮衰老机制将是一大突破，并为预防衰老相关疾病提供新的信息。

线粒体功能障碍和DNA甲基化与衰老相关。GRIM19是线粒体呼吸链复合物I组成成分。我们发现健康人口腔上皮细胞GRIM19部分甲基化，且与年龄有关。本项目分析不同年龄健康人口腔上皮全基因组甲基化和GRIM19启动子甲基化，敲弱 GRIM19的人正常口腔上皮细胞（HNOK）和构建诱导性敲除GRIM19小鼠模型，发现GRIM-19改变导致代谢异常和衰老：SA-β-GAL活性、乳酸产量增加和ROS产生上升，同时，葡萄糖消耗量和NAD+/NADH下降。这些功能改变，可能与GRIM-19敲减导致的SIRT3和FOXO3水平降低，AMPK升高相关。这些结果提示GRIM19对健康者口腔上皮线粒体代谢和衰老信号途径的影响，为预防衰老相关疾病提供新的信息。