基于IL10和IL23R-IL12RB2非编码区多态性介导增强子活性的Behcet病遗传发病机制研究

Behcet病是我国最常见的致盲性葡萄膜炎的类型之一，遗传因素在其中起着重要作用。我们近期全基因组关联研究（GWAS）发现了与Behcet病相关的IL10和IL23R-IL12RB2两个重要基因的非编码区的单核苷酸多态性（SNPs），通过GWAS后注释筛选了与该病相关的9个候选功能性SNPs位点以及其靶基因；运用电泳迁移率测定、染色质免疫沉淀、荧光素酶报告基因和CRISPR-Cas9基因编辑等多种分子生物学方法，探讨和确定候选功能性SNPs位点是否位于增强子元件以及其不同等位基因对增强子活性的影响；探讨和确定这些候选SNPs位点不同等位基因对转录因子结合的影响以及其结合转录因子后对增强子活性的影响；探讨和确定它们影响靶基因表达及其参与疾病发生的分子生物学机制。通过上述几个方面的研究将可能阐明非编码区增强子元件上的多态性在Behcet病的作用及其机制，有望为此病的治疗寻找到新靶点。

Behcet’s disease (BD) is one of the most common blinding uveitis in China. Previous reports have shown that genetic factors are associated with this disease, although the exact genetic pathogenesis is unclear. This study was designed to investigate the biological mechanisms explaining the role of disease-associated single nucleotide polymorphisms (SNPs) in the IL-10 and IL23R-IL12RB2 non-coding regions in the pathogenesis of BD. Post-GWAS annotation will be conducted to screen functional candidate SNPs (rs3024490, rs1800871, rs3021094, rs1518110, rs1554286, rs1518111, rs1800872, rs72676067, and rs4655686) and target genes related to the disease. Subsequently, we will determine whether these candidate SNPs are located in enhancer elements, and we will examine the allele-specific effects on enhancer activities. Then we will investigate the effects of these SNPs on the transcriptional factors and the effects of combining risk or non-risk alleles with the transcriptional factors on enhancer activities. In addition, the molecular mechanisms of candidate SNPs acting on the expression of target genes and getting involved in pathogenesis will be further investigated and confirmed. It will be possible to clarify the role and mechanism of non-coding region SNPs in BD through our conclusions in this study and to find new targets for the treatment of this disease.