Linc00665竞争性结合miR-142-5p调控CUL4B表达对Graves病CD4+T细胞的作用及机制研究

Graves' Disease (GD) is a common autoimmune disease, and CD4+T cell dysfunction has always been a scientific issue of international concern. In the preliminary study, using gene chip technology, bioinformatics prediction and expanded population sample verification, we detected that Linc00665 expression in peripheral blood CD4+T cells of patients with initial GD was increased and negatively correlated with miR-142-5p and positively correlated with serum FT3, FT4 and TRAb and that CUL4B may be a potential target gene of miR-142-5p. However, there is no related reports about the function and mechanism of Linc00665 in GD. Therefore, we intend to choose the technique of flow cytometry, CRISPR/Cas9, luciferase reporter gene, and other advanced techniques, so as to demonstrate that Linc00665 in CD4+T cells may function as a competing endogenous RNA combined with miR-142-5p to regulate CUL4B of the potential target gene of miR-142-5p through the perspectives of population sample, tissue cells, and molecular level, which will provide new ideas for the pathogenesis and clinical therapeutics of GD.

Graves病（GD）是一种常见的自身免疫性疾病，CD4+T细胞功能异常一直是国际前沿关注的科学问题。我们前期通过基因芯片技术、生物信息学预测及扩大样本验证显示，初发GD病人外周血CD4+T细胞Linc00665表达升高及miR-142-5p下降，二者呈负相关，Linc00665的表达与血清FT3、FT4及TRAb水平正相关，且CUL4B可能是miR-142-5p的潜在靶基因。然而其机制不清，国内外未见研究报道。因此，本课题拟采用流式细胞术、CRISPR/Cas9、荧光素酶报告基因等技术，从人群样本、组织细胞、分子水平等多层面明确Linc00665在CD4+T细胞的作用，论证Linc00665通过结合miR-142-5p发挥ceRNA作用，调控miR-142-5p潜在靶基因CUL4B表达的机制，为GD发病机制及临床诊疗提供新思路。

Graves病（Graves’ disease，GD）是临床上引起甲状腺功能亢进症最常见的原因，其发病机理尚不完全清楚。研究表明，CD4+辅助性T细胞（Th）的失衡与GD发病密切相关，Th细胞包括Th1、Th2、Th17、调节性T细胞（Treg）等。Treg和Th17细胞的失衡，在自身免疫性疾病的发生发展中发挥重要作用。本课题组通过广泛阅读文献、高通量mRNA测序及扩大样本验证等筛选出NR4A2基因。有研究显示，NR4A2主要表达于Treg细胞，可促进CD4+T细胞向Treg细胞分化。但是NR4A2在GD中的表达及其对CD4+T细胞作用尚不清楚。本研究采用流式细胞术、qRT-PCR、免疫印迹等技术，从人群样本、组织细胞及分子水平研究NR4A2在初发GD患者CD4+T细胞中的表达变化，探索低表达的NR4A2引起GD患者Treg、Th17失衡的机制，为完善GD发病机制、挖掘临床治疗新靶点提供理论依据。

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