mTOR信号通路在口腔扁平苔藓T细胞免疫应答中的作用研究

Oral lichen planus (OLP) is a chronic T cell-mediated immunity and inflammation disease. Mammalian target of rapamycin (mTOR) plays a key role in antigen presentation, T cell immunity, and immune responses in keratinocyte. Thus, it is speculated that dysregulated mTOR signaling in T cell might involve in OLP through directing T cell homeostatic and functional fates, and in further regulating keratinocyte biology. (1) To detect mTOR signaling in T cell of OLP using PCR arrays, and validate the altered expression of selected genes at mRNA and protein level. (2) To examined the activation level of mTOR signaling in OLP, and to analyze the clinical significance. (3) To silence the altered mTOR signaling via shRNA, and then examine their effects on T cell metabolism, differentiation, activation, proliferation, trafficking, apoptosis and autophagy. (4) To test the role of mTOR signaling in crosstalk between T cell and keratinocyte in OLP. (5) To investigate the effect of mTOR inhibition on OLP T cell immune function. (6) To assay the regulation of metabolic antagonists on T cell immune function mediated by mTOR signaling in OLP. From the perspective of T cell immune and metabolism, this subject potentially reveals an important pathological window for OLP and provides novel strategies for targeted therapeutic intervention of OLP, which not only has high theoretical significance, but also has important practical value.

口腔扁平苔藓（OLP）是T细胞介导的慢性免疫性炎症疾病，mTOR信号通路可调控抗原提呈、T细胞免疫应答和角质形成细胞的免疫反应。本课题提出OLP中存在着异常的mTOR信号，这种异常表达通过影响T细胞的平衡和免疫功能，参与OLP的发生发展。（1）PCR芯片筛选OLP T细胞差异表达的mTOR通路分子。（2）研究OLP中mTOR通路分子的活化水平及其临床意义。（3）采用RNA干扰技术沉默T细胞上差异表达的mTOR信号分子，观察T细胞的代谢、分化、活化、增殖、凋亡、迁移、自噬情况。（4）研究mTOR信号在T细胞和角质形成细胞交互对话中的作用。（5）探究mTOR通路抑制剂对OLP T细胞免疫功能的调控效应。（6）分析代谢抑制剂对mTOR介导的T细胞免疫功能的调节作用。本课题从T细胞免疫和代谢的全新角度为研究OLP的免疫机制及探索OLP的靶向治疗提供实验依据，具有重要的理论意义和应用价值。

口腔扁平苔藓（oral lichen planus，OLP）是一种由T细胞介导的慢性免疫炎性疾病。哺乳动物雷帕霉素靶蛋白（mammalian target of rapamycin，mTOR）可以整合免疫信号和代谢信号，参与调控T细胞的代谢、增殖、凋亡和分化。本项目旨在从代谢与免疫双重角度研究OLP T细胞中mTOR信号通路的作用和调控机制。.主要研究内容包括（1）检测 OLP 患者外周血 T细胞及病损组织中mTOR信号通路相关基因及蛋白的差异表达，并分析其与OLP临床特点之间的关系。（2）研究mTOR信号通路对T细胞的葡萄糖代谢、增殖、凋亡及亚群分化的影响。（3）探究不同mTOR信号通路上游抑制剂对T细胞免疫功能的影响。（4）分析葡萄糖及葡萄糖代谢抑制剂2-DG对T细胞免疫功能的影响。. 研究结果显示（1）MTOR、4E-BP1、PLD2、HIF1α在糜烂型OLP外周T细胞及局部病损组织中的表达降低，PLD2、Hif1α在糜烂型OLP病损组织T细胞浸润带的表达升高。（2）抑制mTOR活性能够降低T细胞的葡萄糖摄取率，减弱GLUT1、LDHA、PDH的表达，抑制T细胞增殖，促进其凋亡，并调控亚群分化平衡。（3）mTOR通路抑制剂能够抑制T细胞增殖，促进T细胞凋亡，调控亚群分化平衡。（4）葡萄糖可影响T细胞的糖酵解、增殖、凋亡及亚群分化等免疫功能。（5）糖酵解抑制剂2-DG能够抑制T细胞增殖，促进T细胞凋亡, 调控T细胞亚群分化平衡。.本研究提示OLP外周血T细胞和病损组织中存在mTOR信号通路的异常表达，且在一定程度上可评估OLP的疾病严重程度。mTOR能够促进T细胞的糖代谢与增殖，抑制其凋亡, 调控T细胞亚群分化平衡，进而介导T细胞免疫应答，参与OLP的发病机制，因此靶向mTOR通路可抑制T细胞免疫应答，对治疗OLP有潜在的应用价值。糖酵解抑制剂2-DG对治疗OLP亦有潜在的应用价值。