Human Genetics and Clinical Translation

项目摘要 青少年特发性脊柱侧凸（AIS）是一种脊柱扭曲的情况，是最常见的儿科疾病， 肌肉骨骼疾病，影响全球3%的儿童。患有AIS的儿童有严重毁容的风险，背部 疼痛和肺功能障碍需要治疗AIS的女孩人数超过男孩， 五倍AIS的治疗是手术治疗，而不是预防性的，因为潜在的病因是未知的。 在美国，AIS的医院费用每年超过10亿美元，而且增长速度明显快于 用于其他儿科手术。我们的总体目的是了解AIS的生物学原因， 早期诊断、预防和非侵入性生物治疗。AIS是一种复杂的遗传性疾病。基因组 我们小组和其他人的广泛关联研究（GWAS）已经确定了AIS相关区域， 推测的调节序列。例如，与我们的项目3（基因组学）合作者，我们最近 表明AIS相关的变异破坏了PAX 1基因的一个假定的增强子， 在脊椎发育中。我们还证明了该位点与女性AIS特异性相关。 虽然这些GWAS已经产生了值得进一步研究的位点，但它们累积起来占总数的<5 AIS的遗传风险在这个项目中，我们提出了新的方法来识别基因和突变， 带来重大疾病风险。在一种方法中，我们将在我们的收集中进行外显子组聚焦的GWAS。 2，750例AIS病例和> 20，000例人群对照，以发现AIS候选者中的疾病相关突变 基因及其调控元件。在我们的第二种方法中， (WGS)与受影响的男性进行三人组研究，以发现预期会带来强烈疾病风险的突变。选择 基因和调控序列将通过斑马鱼的基因组编辑进一步表征（项目2）， 增强子研究（项目3）。我们将通过大规模的基因重测序来确定AIS的突变负担。 GWAS在该项目中发现的候选基因和调控区域，以及 项目2和3。例如，我们的项目2（斑马鱼）合作者发现， kif 6在斑马鱼中产生AIS表型，促使我们将该基因添加到我们的候选列表中。为我们 我们将使用我们建立的方法对至少4,000例AIS病例和对照进行重新测序， 基于分子倒置探针的靶向和大规模平行测序。提供关键试剂， 假设驱动的AIS功能分析，我们将同时扩大我们现有的DNA，细胞， 组织和来自儿科整形外科中心确定的患者的手术样本。最后，对于患者 我们将与AIS的临床专家合作，评估潜在的表型 可以定义临床亚型的相关性。通过与计划中的其他项目协同，我们将定义一个 AIS遗传风险的很大一部分，并建立重要的工具来表征潜在的 疾病机制。

Project Summary Adolescent idiopathic scoliosis (AIS) is a twisting condition of the spine and is the most common pediatric musculoskeletal disorder, affecting 3% of children worldwide. Children with AIS risk severe disfigurement, back pain, and pulmonary dysfunction later in life. Girls requiring treatment for AIS outnumber boys by more than five-fold. AIS is treated symptomatically rather than preventively because the underlying etiology is unknown. Hospital charges for AIS surpass one billion dollars annually in the U.S. and are rising significantly faster than for other pediatric procedures. Our overall purpose is to understand the biologic causes of AIS as a means to early diagnosis, prevention and non-invasive biologic treatment. AIS is a complex genetic disease. Genome wide association studies (GWAS) by our group and others have identified AIS-associated regions harboring presumed regulatory sequences. For example, with our Project 3 (Genomics) collaborators we have recently shown that AIS-associated variants disrupt a putative enhancer of the PAX1 gene that is known to participate in spinal development. We also demonstrated that this locus is specifically associated with AIS in females. While these GWAS has yielded loci worthy of further study, they cumulatively account for <5% of the total genetic risk in AIS. In this Project we propose new approaches to identify the genes and mutations expected to convey substantial disease risk. In one approach we will perform exome-focused GWAS in our collection of 2,750 AIS cases and >20,000 population controls to discover disease-associated mutations in AIS candidate genes and their regulatory elements. In our second approach we will perform whole genome sequencing (WGS) in trios with affected males to discover mutations expected to convey strong disease risk. Selected genes and regulatory sequences will be characterized further by genome editing in zebrafish (Project 2) and enhancer studies (Project 3). We will define the mutational burden in AIS by large-scale re-sequencing of candidate genes and regulatory regions discovered in this project by GWAS, as well as those discovered by in Projects 2 and 3. For example, our Project 2 (Zebrafish) collaborators have discovered that multiple alleles of kif6 produce an AIS phenotype in zebrafish, prompting us to add this gene to our candidate list. For our mutation screens we will re-sequence at least 4,000 AIS cases and controls using our established method of molecular inversion probe-based targeting and massively parallel sequencing. To provide critical reagents for hypothesis-driven functional analysis of AIS, we will simultaneously expand our existing biobank of DNA, cells, tissues, and surgical samples from patients ascertained in pediatric orthopaedic centers. Finally, for patients with defined AIS-causing mutations, we will team with clinical experts in AIS to evaluate potential phenotypic correlations that may define clinical subtypes. By synergizing with other projects in the program we will define a substantial fraction of the genetic risk in AIS and establish important tools for characterizing the underlying disease mechanisms.