Identification and characterization of noncoding mutations causing autoinflammatory diseases

自身炎症性疾病，也称为周期性发热综合征，是由一组单基因或复杂的遗传性疾病定义的，其特征在于在缺乏高水平自身抗体或抗原特异性T细胞的情况下似乎无缘无故的炎症发作。全基因组关联研究（GWAS）和全基因组测序（WGS）已成功应用于将广泛的单核苷酸多态性（SNP）与自身炎症性疾病显著关联。有趣的是，GWAS鉴定的疾病相关变异中只有大约10%实际上属于基因编码区，而超过90%与基因组的非编码区一致，因此可能是调控区。前瞻性研究的目标是使用GWAS和WGS数据与公共基因组和表观基因组数据集相结合，以识别位于细胞类型特异性增强子内的自身炎症疾病相关SNP。然后通过先天免疫细胞如巨噬细胞或树突细胞中的结构和功能测定来确认候选增强子。此外，CRISPR/Cas9技术将用于删除完整的增强子片段或将特定的SNP引入树突状细胞和斑马鱼的基因组中，这是一种研究免疫系统缺陷的成熟动物模型。随后，转录组分析提供了通过修饰的增强子调控的候选基因的信息。所描述的研究将有助于理解增强子对人类免疫生物学的意义，并获得对增强子的作用以及这些增强子中SNP对单基因和复杂自身炎症性疾病的表现和发展的影响的新的机制见解。

Autoinflammatory diseases, also called periodic fever syndromes, are defined by a group of monogenic or complex genetic disorders characterized by seemingly unprovoked episodes of inflammation in the absence of high levels of auto-antibodies or antigen-specific T cells. Genome-wide association studies (GWAS) and whole genome sequencing (WGS) were successfully applied to significantly associate a wide range of single nucleotide polymorphisms (SNPs) to autoinflammatory diseases. Interestingly, only approximately 10% of the disease-associated variants identified by GWAS actually fall into gene coding regions, while more than 90% coincide with noncoding and thus potentially regulatory regions of the genome. The goal of the prospective study is to use GWAS and WGS data in combination with public genomic and epigenomic datasets to identify autoinflammatory disease-associated SNPs that are located within cell-type specific enhancers. The candidate enhancers are then confirmed through structural and functional assays in innate immune cells such macrophages or as dendritic cells. Additionally, CRISPR/Cas9 technology will be used to either delete the complete enhancer fragment or to introduce a specific SNP into the genome of dendritic cells and zebrafish, a well established animal model to study defects of the immune system. Subsequently, transcriptome analyses provide information of candidate genes regulated through the modified enhancers. The described study will help to understand the significance of enhancers on human immunobiology and to gain new mechanistic insights into the role of enhancers and the effect of SNPs within these enhancers for manifestation and development of monogenic and complex autoinflammatory diseases.