Investigating the contribution of non-coding genetic variation to rare disorders

项目摘要/摘要 基因检测正在迅速崛起，成为医学的基石，使有针对性的治疗成为可能 常见和罕见的疾病，并使患者、家庭和社区了解 他们的状况。然而，尽管在过去十年中取得了重大进展，包括引入了 临床全基因组测序，临床遗传学的研究在很大程度上仍然局限于不到1%的 基因组，外显子组。这一限制被认为是临床基因组测序 未能为大多数推定的患者阐明致病元变异体(S) 接受检测的单基因疾患。据推测，这些患者中有相当大比例 破坏已知孟德尔病毒基因调控结构的致病非编码变异体 基因，一类变异体，使用当前的测序方法解释得很差。直接解决这一问题 限制要求对执行和解释基因测试的方式进行大规模修订，如 这项提议。具体地说，这项建议旨在通过以下方式克服人类遗传学的这一根本限制 利用我们最近开发的一种新方法来同时绘制遗传和表观遗传学 样本的景观，从而阐明了非编码基因变异对功能的影响 局部染色质结构和基因调控模式--全外膜基因组测序(WEGS)。vbl.使用 通过这种方法，我们计划直接测试这一假设，即罕见的非编码基因改变有助于 单基因疾病。在目标1中，我们将使用WEGS方法来表征 健康个体和已知疾病患者的非编码序列、结构和表观遗传学改变 印记障碍。这一目标的目的是建立WEGS的灵敏度和识别能力 破坏当地染色质结构和基因调控模式并改善我们的 了解非编码遗传变异对功能的影响。在目标2中，我们将直接评估 应用WEGS研究罕见的非编码基因改变对单基因疾病的作用 怀疑为单基因疾病的患者，其整个外显子组或基因组测序以前都不是 诊断。总体而言，这项提议有可能极大地改变我们处理基因组测试的方式 以及我们对非编码序列和结构变异对基因调控影响的理解 模式和人类疾病。

PROJECT SUMMARY/ABSTRACT Genetic testing is rapidly emerging as a cornerstone of medicine, enabling targeted therapies for patients with both common and rare disorders, and empowering patients, families and communities with knowledge about their condition. However, despite significant advances over the past decade, including the introduction of clinical whole genome sequencing, clinical genetics remains largely limited to the study of less than 1% of the genome, the exome. This limitation is thought to underlie the fact that clinical genomic sequencing is unsuccessful in elucidating the culprit pathogenic variant(s) for the majority of patients with presumed monogenic disorders who undergo testing. It is hypothesized that a significant percentage of these patients harbor pathogenic non-coding variants that disrupt the gene regulatory architecture of known Mendelian genes, a class of variants poorly illuminated using current sequencing approaches. Directly addressing this limitation requires a wholesale revision of how genetic testing is performed and interpreted, as is outlined in this proposal. Specifically, this proposal aims to overcome this fundamental limitation of human genetics by leveraging a novel approach we recently developed for simultaneously mapping the genetic and epigenetic landscape of a sample, thereby illuminating the functional impact of non-coding genetic variants that disrupt local chromatin architecture and gene regulatory patterns – Whole Epi-Genome Sequencing (WEGS). Using this approach, we plan to directly test the hypothesis that rare non-coding genetic alterations contribute to monogenic disorders. In Aim 1, we will use the WEGS approach to characterize the gene regulatory impact of non-coding sequence, structural and epigenetic alterations in healthy individuals as well as patients with known imprinting disorders. The goal of this aim is to establish the sensitivity and power of WEGS for identifying genetic variants that disrupt local chromatin architecture and gene regulatory patterns and improve our understanding of the functional impact of non-coding genetic variation. In Aim 2, we will directly evaluate the contribution of rare non-coding genetic alterations to monogenic disorders by applying WEGS to patients with suspected monogenic disorders for whom whole exome or genome sequencing has previously been non- diagnostic. Overall, this proposal has the potential to dramatically change how we approach genomic testing and our understanding of the impact of non-coding sequence and structural variation on gene regulatory patterns and human disease.