An Ultraconserved Brain-Specific Enhancer Within ADGRL3 (LPHN3) Underpins Attention-Deficit/Hyperactivity Disorder Susceptibility.
An Ultraconserved Brain-Specific Enhancer Within ADGRL3 (LPHN3) Underpins Attention-Deficit/Hyperactivity Disorder Susceptibility.
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DOI：
10.1016/j.biopsych.2016.06.026
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发表时间：
2016-12-15
影响因子：
10.6
通讯作者：
Muenke M
中科院分区：
文献类型：
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作者：
Martinez AF;Abe Y;Hong S;Molyneux K;Yarnell D;Löhr H;Driever W;Acosta MT;Arcos-Burgos M;Muenke M
遗传因素容易导致注意力缺陷/多动障碍（ADHD）。先前的研究报道了 ADGRL3 内基因变异与 ADHD 的联系和关联。在这项研究中，我们对该基因中的非编码变异进行了功能分析，将其视为可能的病理因素。使用计算机、体外和体内方法来识别和表征 ADGRL3 连锁区域 (~207 Kb) 内的进化保守元件。对 838 名个体（372 名受影响者和 466 名未受影响者）进行的基于家族的遗传分析发现，其中一些保守元件中含有与 ADHD 相关的 SNP。荧光素酶测定和斑马鱼 GFP 转基因测试了转录增强子活性的保守元件。电动迁移率测定用于验证 ADHD 风险等位基因对转录因子结合的破坏。发现了一个具有转录增强子功能的超保守元件（ECR47）。 ECR47 中由 rs17226398、rs56038622 和 rs2271338 形成的三变异 ADHD 风险单倍型，使神经母细胞瘤和星形细胞瘤细胞中的增强子活性降低 40% (PBonferroni<0.0001)。该增强子还以组织特异性方式驱动斑马鱼大脑中的 GFP 表达，共享内源 ADGRL3 表达的方面。 rs2271338 风险等位基因破坏 YY1 的结合，YY1 是中枢神经系统发育和功能的重要因素。对死后人脑组织的表达数量性状位点分析揭示了 rs2271338 与丘脑中 ADGRL3 表达减少之间的关联。这些结果揭示了常见非编码变异的第一个功能证据，对 ADHD 的病理学具有潜在影响。
Genetic factors predispose to attention deficit/hyperactivity disorder (ADHD). Previous studies have reported linkage and association to ADHD of gene variants within ADGRL3. In this study, we functionally analyzed non-coding variants in this gene as likely pathological contributors. In silico, in vitro and in vivo approaches were used to identify and characterize evolutionary conserved elements within the ADGRL3 linkage region (~207 Kb). Family-based genetic analyses on 838 individuals (372 affected and 466 unaffected) identified ADHD-associated SNPs harbored in some of these conserved elements. Luciferase assays and zebrafish GFP transgenesis tested conserved elements for transcriptional enhancer activity. Electromobility shift assays were used to verify transcription factor binding disruption by ADHD risk alleles. An ultraconserved element was discovered (ECR47) that functions as a transcriptional enhancer. A three-variant ADHD risk haplotype in ECR47, formed by rs17226398, rs56038622 and rs2271338, reduced enhancer activity by 40% in neuroblastoma and astrocytoma cells (PBonferroni<0.0001). This enhancer also drove GFP expression in the zebrafish brain in a tissue-specific manner, sharing aspects of endogenous ADGRL3 expression. The rs2271338 risk allele disrupts binding of YY1, an important factor in the development and function of the central nervous system. Expression quantitative trait loci analysis of post-mortem human brain tissues revealed an association between rs2271338 and reduced ADGRL3 expression in the thalamus. These results uncover the first functional evidence of common non-coding variants with potential implications for the pathology of ADHD.