雷帕霉素联合芬戈莫德通过Akt-mTOR和ERK通路调控实验性自身免疫性脑脊髓炎Foxp3基因甲基化

Background Multiple sclerosis (MS) is one of the important diseases which cause young people disabled, lack of effective treatments. Experimental autoimmune encephalomyelitis (EAE) is the best mouse model for studying MS. Question Large dose of rapamycin or fingolimod, respectively, may be effective for EAE. But the clinical application is limited. Could the combination of subclinical doses alleviate the pathological damage of EAE? Hypothesis We put forward a hypothesis that the combination of rapamycin and fingolimod could act synergistically and modulate immune response, and regulate Foxp3 demethylation by complementary immune mechanism. Means Our previous study found that fingolimod could treat EAE in a dose-dependent manner and regulate Th subsets function and differentiation. As a series of research, this project intends to adopt suboptimal doses of rapamycin and fingolimod to prevent EAE, evaluating its functions and differentiation of Th subsets. We observed the influence of joint proposal of the activity of Akt-mTOR and ERK pathway and the methylation level of Foxp3 gene in Treg cells, in order to make clear the targets of effective treatment. Results This study might prove the synergistic effect of combination of rapamycin and fingolimod, and further explore the roles of the epigenetics of Foxp3 gene in pathogenesis of EAE and treatment mechanism. This study will provide basis for accurate treatment of MS.

背景 多发性硬化(MS)是致青年人残疾的重要疾病，无特效治疗，实验性自身免疫性脑脊髓炎(EAE)是研究MS的经典模型。问题 大剂量雷帕霉素和芬戈莫德治疗EAE有效，但临床应用受限，亚剂量联合用药能否阻止EAE病情？假说 本项目提出雷帕霉素联合芬戈莫德治疗EAE具有协同效应，通过免疫互补作用调控Foxp3基因去甲基化，调节免疫应答。方法 前期研究发现芬戈莫德治疗EAE有效，呈剂量依赖性，调节Th亚群功能及分化，做为系列研究，本项目拟采用亚剂量雷帕霉素联合芬戈莫德干预EAE，评估该方案的治疗效果和对Th亚群功能及分化的影响，并通过观察联合方案对Akt-mTOR和ERK通路及Treg细胞Foxp3基因甲基化的影响以明确治疗有效的靶点。预期结果 明确联合用药对EAE有协同效果及免疫互补作用，阐述Foxp3基因的表观遗传学改变在EAE发病及治疗机制中的作用，为临床寻找MS的精准治疗方案提供依据。

背景 多发性硬化(MS)是致青年人残疾的重要疾病，但是其免疫学发病机制并不明确，实验性自身免疫性脑脊髓炎(EAE)是研究MS的经典动物模型。主要研究内容 本课题通过成功诱导EAE小鼠模型，评估不同干预组EAE小鼠的症状评分和病理组织学改变，明确雷帕霉素和芬戈莫得亚剂量联合用药的预防治疗效果；分析亚剂量联合用药对EAE小鼠Th17/Treg细胞的分化和功能产生的影响；阐述亚剂量联合用药对EAE小鼠Treg细胞中Foxp3基因甲基化水平的调控作用；探讨联合用药作用于EAE小鼠的免疫互补机制。重要结果 雷帕霉素和芬戈莫德两种药物亚剂量联合应用能够缓解EAE，减少中枢神经系统炎症细胞浸润，调节Th17/Treg细胞平衡，通过Akt-mTOR和ERK通路调控Treg细胞Foxp3基因甲基化，具有互补免疫调节作用。关键数据 两种药物亚剂量联合应用能够减少Th17的分化和功能，增加Treg细胞的分化和功能，通过抑制Akt-mTOR和ERK通路导致Treg细胞Foxp3基因去甲基化。科学意义 阐述了亚剂量联合用药治疗EAE的免疫学互补机制，为寻找EAE/MS的精准治疗靶点提供依据。