同型半胱氨酸经SNF5介导组蛋白修饰与DNA甲基化交互作用调控TSLP致动脉粥样硬化的分子机制

Atherosclerosis (As) is a chronic inflammatory disease, and its occurrence and development are closely related with immunoinflammatory reactions. It has not been reported whether TSLP, the common upstream signaling molecule of DC and Treg functional regulation, involved in Hcy induced As. Team’s research has confirmed SNF5 and TSLP are the key genes of Hcy induced As and histone modification and DNA methylation are important mechanism for transcriptional regulation of genes. However, the mechanism of homocysteine induced atherosclerosis by regulating TSLP through SNF5 has not been elucidated. Therefore, this project proposed by GST-pulldown and CHIP to determine the distribution and interactions of SNF5 and TSLP in Hcy-induced As. Therefore, MeDIP-qPCR is used to analyze the changes of H3K9 methylation and histone acetylation and TSLP DNA methylation, to illuminate the regulatory mechanism of histone modification and DNA methylation in the formation of AS. We will adopt blocking SNF5, DNMT1, SUV39H1 and HDAC1 and other strategies to investigate the effect on the target genes after the respective balance model are broken; Reveal SNF5 through histone modification and DNA methylation interaction regulates TSLP-mediated immune inflammatory response mechanisms. We hope this study can provide the theory basis for AS target treatment.

动脉粥样硬化(As)是一种慢性炎症性疾病，其发生发展与免疫炎症反应关系密切，TSLP作为DCs和Treg功能调节共同的上游信号分子，是否参与同型半胱氨酸(Hcy)致As未见报道。前期观察到SNF5和TSLP是Hcy致As关键基因，而组蛋白修饰和DNA甲基化是基因转录调控的重要机制，但其在Hcy经SNF5调控TSLP致As的机制尚未阐明。因此本课题拟通过GST-pulldown实验和CHIP技术等明确SNF5和TSLP在Hcy致As中的分布及相互作用；采用 MeDIP-qPCR分析H3K9甲基化和乙酰化与TSLP DNA甲基化的变化，阐明组蛋白修饰与DNA甲基化在As形成中调控机制；采用阻断SNF5、DNMT1、SUV39H1和HDAC1等策略探讨各自平衡模式被打破后对靶基因的影响，揭示SNF5经组蛋白修饰与DNA甲基化交互作用调控TSLP介导免疫炎症反应机制，为As靶向治疗提供理论依据。

动脉粥样硬化(As)是一种慢性炎症性疾病，其发生发展与免疫炎症反应关系密切。细胞因子白介素-1β（interleukin-1β，IL-1β）是机体内最重要的几种促炎细胞因子之一，主要来自于受到各种刺激、活化后的单核-巨噬细胞。有证据表明IL-1β参与了动脉粥样硬化炎症反应。此外，课题组前期观察到染色质重塑复合物SWI/SNF的核心亚基SNF5（Sucrose Non-Fermentable 5, SNF5）是Hcy致As关键基因，而组蛋白修饰是基因转录调控的重要机制。但是，其在Hcy致As的发生发展过程中如何调控IL-1β并最终引起下游变化的机制尚不清楚。因此本课题旨在分析SNF5在Hcy促进As中所扮演的角色，明确IL-1β为Hcy促进动脉粥样硬化中SNF5的靶基因，旨在揭示SNF5通过组蛋白H3K4甲基化调控IL-1β从而促进动脉粥样硬化的具体机制，为动脉粥样硬化靶向治疗提供理论依据。