小鼠结肠上皮细胞在免疫刺激环境下的表观遗传学研究

Our research is based on the purified mice colon epithelial cells, which are stimulated by kinds of factors including natural bacterial products LPS, MDP and IL-17 family cytokines IL-17A, IL-17C, and IL-17E. RNA-Seq is used for testing the mRNA expression difference after stimulation, MeDIP-Seq is used for testing the DNA demethylation level after stimulation, and ChIP-Seq is used for histone modification. We combined the gene expression and epigenetic modification by analyzing the sequencing data. The DNA demethylation mechanisms are investigated using Tet KO mice, and the commensal bacteria dependence would be investigated using Germ Free mice. Furthermore, the mechanism of IL-17 function would be elucidated, mainly focusing on the IL-17-IL-17R-Tet mediated epigenetic changes. Meanwhile the colon naïve T cells are isolated for Th17 differentiation, then effect of the gut lymphocytes on epithelial cells is observed. Finally the project would find novel targets and drugs for IBD.

采用不同因子刺激小鼠结肠上皮细胞，包括天然细菌产物LPS, MDP，以及IL-17家族细胞因子包括IL-17A, IL-17C, IL-17E，综合分析RNA-Seq等二代测序结果，将基因表达与基因表观遗传修饰结合起来，筛选相关差异基因，并进行PCR验证。利用Tet敲除小鼠研究差异基因的去甲基化调控机制，Germ Free小鼠研究基因的去甲基化是否依赖肠道共生菌的作用。进一步阐明IL-17类细胞因子的作用机制，主要研究IL-17-IL-17R-Tet信号通路对基因的表观遗传调控作用。同时提取结肠Naïve T细胞，诱导细胞分化为Th17细胞，观察肠道淋巴细胞分化对上皮细胞基因表观遗传的影响。该课题为找到炎性肠病相关的基因靶点及药物提供新的思路与坚实的基础。