Verlaan Dominique J, Berlivet Soizik, Hunninghake Gary M, Madore Anne-Marie, Larivière Mathieu, Moussette Sanny, Grundberg Elin, Kwan Tony, Ouimet Manon, Ge Bing, Hoberman Rose, Swiatek Marcin, Dias Joana, Lam Kevin C L, Koka Vonda, Harmsen Eef, Soto-Quiros Manuel, Avila Lydiana, Celedón Juan C, Weiss Scott T, Dewar Ken, Sinnett Daniel, Laprise Catherine, Raby Benjamin A, Pastinen Tomi, Naumova Anna K
McGill University and Genome Quebec Innovation Centre, Quebec, Canada.
Am J Hum Genet. 2009 Sep;85(3):377-93. doi: 10.1016/j.ajhg.2009.08.007.
Common SNPs in the chromosome 17q12-q21 region alter the risk for asthma, type 1 diabetes, primary biliary cirrhosis, and Crohn disease. Previous reports by us and others have linked the disease-associated genetic variants with changes in expression of GSDMB and ORMDL3 transcripts in human lymphoblastoid cell lines (LCLs). The variants also alter regulation of other transcripts, and this domain-wide cis-regulatory effect suggests a mechanism involving long-range chromatin interactions. Here, we further dissect the disease-linked haplotype and identify putative causal DNA variants via a combination of genetic and functional analyses. First, high-throughput resequencing of the region and genotyping of potential candidate variants were performed. Next, additional mapping of allelic expression differences in Yoruba HapMap LCLs allowed us to fine-map the basis of the cis-regulatory differences to a handful of candidate functional variants. Functional assays identified allele-specific differences in nucleosome distribution, an allele-specific association with the insulator protein CTCF, as well as a weak promoter activity for rs12936231. Overall, this study shows a common disease allele linked to changes in CTCF binding and nucleosome occupancy leading to altered domain-wide cis-regulation. Finally, a strong association between asthma and cis-regulatory haplotypes was observed in three independent family-based cohorts (p = 1.78 x 10(-8)). This study demonstrates the requirement of multiple parallel allele-specific tools for the investigation of noncoding disease variants and functional fine-mapping of human disease-associated haplotypes.
17号染色体12q12 - q21区域的常见单核苷酸多态性(SNPs)会改变患哮喘、1型糖尿病、原发性胆汁性肝硬化和克罗恩病的风险。我们和其他人之前的报告已将疾病相关的基因变异与人类淋巴母细胞系(LCLs)中GSDMB和ORMDL3转录本的表达变化联系起来。这些变异还会改变其他转录本的调控,这种全结构域的顺式调控效应提示了一种涉及长程染色质相互作用的机制。在此,我们进一步剖析与疾病相关的单倍型,并通过遗传和功能分析相结合的方法鉴定出可能的因果DNA变异。首先,对该区域进行高通量重测序以及对潜在候选变异进行基因分型。接下来,对约鲁巴人单倍型图谱LCLs中等位基因表达差异进行额外定位,使我们能够将顺式调控差异的基础精细定位到少数几个候选功能变异上。功能分析确定了核小体分布中的等位基因特异性差异、与绝缘子蛋白CTCF的等位基因特异性关联以及rs12936231的弱启动子活性。总体而言,本研究显示一个常见疾病等位基因与CTCF结合及核小体占据情况的变化相关联,从而导致全结构域顺式调控改变。最后,在三个独立的基于家系的队列中观察到哮喘与顺式调控单倍型之间存在强关联(p = 1.78 x 10(-8))。本研究证明了需要多种平行的等位基因特异性工具来研究非编码疾病变异以及对人类疾病相关单倍型进行功能精细定位。