LncRNA PRKG1-AS1调控HMGB1/PRKG1参与Th17细胞免疫反应在Vogt-小柳原田综合征中的分子机制研究

Vogt-小柳原田(VKH)综合征是我国常见且致盲率高的葡萄膜炎类型，其核心免疫反应的调控机制尚不完全清楚。我们前期RNA测序发现对Th17细胞具有潜在调控作用的lncRNA PRKG1-AS1在VKH活动期表达上调，结合生信及预实验结果，提出科学假设“PRKG1-AS1可通过顺式作用调控邻近基因PRKG1，并内源性竞争miRNA-20a-5p，协同激活HMGB1/PRKG1，继而影响Th17细胞免疫反应而参与VKH发病”。项目拟用qRT-PCR、ELISA、流式、转染、双荧光素酶报告等技术，首先探讨PRKG1-AS1在VKH患者PBMCs、CD4+T细胞及血清中的表达规律；其次探讨PRKG1-AS1对Th17细胞免疫反应的影响；最后探讨PRKG1-AS1对PRKG1、miR-20a-5p的调控方式和对HMGB1/PRKG1的影响，以期阐明其在VKH中的作用及调控机制，为防治提供新靶点。

Vogt-Koyanagi-Harada (VKH) syndrome is a common uveitis entity in China with high incidence rate and blinding rate，however, the regulatory mechanism of its core pathogenic immune response remains unclear. Using next-generation RNA sequencing and validation experiments, we found that the expression of lncRNA PRKG1-AS1, a potential LncRNA with immunoregulation function on Th17 cell, was increased in VKH patients with active uveitis. Combined with the results of bioinformatics analysis, we propose a hypothesis that "PRKG1-AS1 could not only influence the translation of its neighboring gene PRCK1 via cis regulation, but also could competitively bind to endogenous miR-20a-5p to regulate HMGB1 and PRGK1, which could affect the immune response of Th17 cells in the parthenogenesis of VKH". Quantitative RT-PCR, ELISA, flow cytometry, gene transfection and double luciferase report experiment would be applied in this study. Firstly, the expression of PCKG1-AS1 in PBMC, CD4+ T cells and serum of VKH patients would be tested. Secondly, the influences of PRKG1-AS1 on the T cells immune response involved in the development of VKH would be explored. Then, in order to clarify the mechanism of PRKG1-AS1 in the pathogenesis of VKH, the regulatory model between PRKG1-AS1, PRKG1 and mir-20a-5p would be validated, and the effect of PRKG1-AS1 on HMGB1-PRGK1 pathway would be researched. This project would possibly provide a new target for the clinical prediction and treatment of uveitis.