Division of Endocrinology, Department of Medicine, Mount Sinai Medical Center, New York, New York 10029, USA.
J Biol Chem. 2011 Sep 9;286(36):31168-79. doi: 10.1074/jbc.M111.247510. Epub 2011 Jul 12.
Autoimmune thyroid diseases (AITD) arise from complex interactions between genetic, epigenetic, and environmental factors. Whole genome linkage scans and association studies have established thyroglobulin (TG) as a major AITD susceptibility gene. However, the causative TG variants and the pathogenic mechanisms are unknown. Here, we describe a genetic/epigenetic mechanism by which a newly identified TG promoter single-nucleotide polymorphism (SNP) variant predisposes to AITD. Sequencing analyses followed by case control and family-based association studies identified an SNP (-1623A→G) that was associated with AITD in the Caucasian population (p = 0.006). We show that the nucleotide substitution introduced by SNP (-1623A/G) modified a binding site for interferon regulatory factor-1 (IRF-1), a major interferon-induced transcription factor. Using chromatin immunoprecipitation, we demonstrated that IRF-1 binds to the 5' TG promoter motif, and the transcription factor binding correlates with active chromatin structure and is marked by enrichment of mono-methylated Lys-4 residue of histone H3, a signature of active transcriptional enhancers. Using reporter mutations and siRNA approaches, we demonstrate that the disease-associated allele (G) conferred increased TG promoter activity through IRF-1 binding. Finally, treatment of thyroid cells with interferon α, a known trigger of AITD, increased TG promoter activity only when it interacted with the disease-associated variant through IRF-1 binding. These results reveal a new mechanism of interaction between environmental (IFNα) and genetic (TG) factors to trigger AITD.
自身免疫性甲状腺疾病 (AITD) 是由遗传、表观遗传和环境因素之间的复杂相互作用引起的。全基因组连锁扫描和关联研究已经确定甲状腺球蛋白 (TG) 是 AITD 的主要易感基因。然而,导致 TG 变异的原因和发病机制尚不清楚。在这里,我们描述了一种新发现的 TG 启动子单核苷酸多态性 (SNP) 变异通过遗传/表观遗传机制导致 AITD 的机制。通过测序分析,随后进行病例对照和家系关联研究,确定了一个 SNP(-1623A→G)与高加索人群中的 AITD 相关(p=0.006)。我们表明,SNP(-1623A/G)引入的核苷酸取代改变了干扰素调节因子-1(IRF-1)的结合位点,IRF-1 是一种主要的干扰素诱导转录因子。通过染色质免疫沉淀,我们证明了 IRF-1 结合到 5' TG 启动子序列,并且转录因子的结合与活性染色质结构相关,并通过组蛋白 H3 赖氨酸-4 单甲基化残基的富集来标记,这是活性转录增强子的特征。使用报告基因突变和 siRNA 方法,我们证明疾病相关等位基因 (G) 通过与 IRF-1 结合,赋予 TG 启动子更高的活性。最后,用已知触发 AITD 的干扰素 α 处理甲状腺细胞时,只有当它与疾病相关的变体通过 IRF-1 结合相互作用时,才能增加 TG 启动子的活性。这些结果揭示了环境 (IFNα) 和遗传 (TG) 因素之间相互作用触发 AITD 的新机制。