GWAS-driven targeted resequencing of candidate genes to assess the impact of rare and low-frequency coding variation in restless legs syndrome

GWAS 驱动的候选基因靶向重测序，以评估罕见和低频编码变异对不宁腿综合征的影响

• 批准号: 310572679
• 负责人: Dr. Barbara Schormair
• 金额: --
• 依托单位: Neurologische Klinik und Poliklinik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2016
• 资助国家: 德国
• 起止时间: 2015-12-31 至 2017-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/310572679?language=en
• 关键词: GWAS; driven; targeted; resequencing; candidate

Restless legs syndrome (RLS) is a common sleep-related movement disorder that affects up to 10% of the population in western societies. Despite its profound impact on health and quality of life, the underlying pathophysiology is still poorly understood. So far, genome-wide association studies (GWAS) of the disease have been the most promising approach because they provided the first molecular starting points to study the mechanisms at work.However, these GWAS assessed common, mostly non-coding variants, which have only modest effects on the phenotype, and, inherent to their design, only delineated genomic regions of interest (risk loci) instead of directly identifying causal genes and variants. Here, we aim to identify causal low-frequency and rare variants of potentially high impact on the phenotype. We will sequence coding regions of candidate genes selected from risk loci identified in our meta-analysis of the two largest GWAS in samples of European ancestry (unpublished data). Coding regions will be sequenced in 4,500 cases and 4,500 controls using molecular inversion probes (MIPs) to enrich the targeted sequences, followed by next-generation sequencing (NGS). Association analysis will be performed for individual low-frequency variants as well as by aggregating all variants across a gene in gene-based association tests. Studying low-frequency and rare coding variation in RLS has the potential to identify specific high-impact causal variants and genes. The molecular consequences of such variants will be straightforward to analyze in in-vivo and in-vitro functional assays such iPS cells or animal model to elucidate the pathophysiology of RLS. Finally, this facilitates the molecular analysis of a complex disease such as RLS and will deliver new targets for improved therapeutics or even preventive measures.

不宁腿综合征（RLS）是一种常见的睡眠相关运动障碍，影响西方社会高达10%的人口。尽管其对健康和生活质量的影响深远，但其潜在的病理生理学仍然知之甚少。到目前为止，全基因组关联研究（GWAS）一直是最有前途的方法，因为它们提供了研究工作机制的第一个分子起点。然而，这些GWAS评估的是常见的，主要是非编码变异，这些变异对表型只有适度的影响，并且，其设计固有的，仅描绘感兴趣的基因组区域（风险基因座），而不是直接识别致病基因和变体。在这里，我们的目标是确定因果低频和罕见的变异的潜在高影响的表型。我们将对从我们对欧洲血统样本中两个最大GWAS的荟萃分析中确定的风险位点中选择的候选基因的编码区进行测序（未发表的数据）。将使用分子倒置探针（MIP）对4,500例病例和4,500例对照的编码区进行测序，以富集靶序列，然后进行下一代测序（NGS）。将对个体低频变异体进行关联分析，并通过在基于基因的关联检验中聚合基因中的所有变异体进行关联分析。研究RLS中的低频和罕见编码变异有可能识别出特定的高影响力的因果变异和基因。这些变体的分子结果将直接在体内和体外功能测定（如iPS细胞或动物模型）中分析，以阐明RLS的病理生理学。最后，这有助于对复杂疾病（如RLS）进行分子分析，并将为改进的治疗方法甚至预防措施提供新的靶点。