Identification of Genetic Susceptibility in Adolescent Idiopathic Scoliosis

描述（由申请人提供）：青少年特发性脊柱侧凸（AIS）是最常见的儿科脊柱畸形，影响全球约3%的儿童。AIS严重影响美国的国民健康，造成超过10%的患者严重毁容和残疾，每年花费数十亿美元用于治疗。我们的长期目标是确定AIS的遗传基础，这将为疾病的发病机制提供见解。在本申请中，我们提出使用从在主要儿科整形外科中心治疗的AIS患者系统地确定的DNA集合来鉴定AIS易感基因座。我们的目标是通过全基因组关联测试（GWA）在700个AIS家族三人组中发现疾病相关的遗传位点，并在第二个1,400个三人组队列中测试最重要位点的关联复制。第三个目的是评估与AIS易感性相关的拷贝数变异（CNV）。第四个和第五个目标是在第三个独立的队列中筛选最有希望的~2%的复制基因座，该队列包括在整个美国确定的1,000个三重基因组，并在这些研究确定的选定基因中确定致病变异。目标1将利用Illumina HumanCNV 370-Duo系统对超过318，000个标记单核苷酸多态性（tagSNPs）进行基因分型，并询问约11，000个疑似CNV区域。单标记和多标记（单倍型）统计方法将应用于SNP数据。将评估新生突变和常见拷贝数变异。此外，将开发新的统计方法并应用于CNV数据。以这种方式识别的基因座将根据重要性进行排名。将使用类似的统计方法测试排名靠前的SNP基因座的关联性的复制，所述统计方法应用于在1，400个三人组中使用Illumina GoldenGate系统获得的基因型。将利用Taqman和基于Sequenom的平台对第三组1,000个trios中最有希望的重复位点进行基因分型。进一步的研究，包括计算机模拟方法和直接DNA测序，将用于确定疾病相关基因中可能的致病变异。这些研究将揭示AIS易感基因位点在患者人群中具有强烈的影响。这些发现对于未来许多最终针对预防或替代治疗的研究将是重要的。

DESCRIPTION (provided by applicant): Adolescent idiopathic scoliosis (AIS) is the most common pediatric spinal deformity, affecting ~3% of children worldwide. AIS significantly impacts national health in the U.S., creating severe disfigurement and disability for over 10% of patients and costing billions of dollars annually for treatment. Our long-term objective is to identify genetic underpinnings in AIS that will provide insights into disease pathogenesis. In the present application we propose to identify AIS susceptibility loci using DNA collections systematically ascertained from AIS patients treated at major pediatric orthopedic centers. Our aims are to discover disease-associated genetic loci via genome-wide tests of association (GWA) in 700 AIS family trios, and to test replication of association for most significant loci in a second cohort of 1,400 trios. A third aim is to assess copy number variation (CNV) associated with AIS susceptibility. Fourth and fifth aims are to screen most promising ~2% of replicated loci in a third, independent cohort of 1,000 trios ascertained throughout the U.S., and to identify causal variants in selected genes identified by these studies. Aim 1 will utilize the Illumina HumanCNV370-Duo system to genotype over 318,000 tagging single nucleotide polymorphisms (tagSNPs), and to interrogate ~11,000 regions of suspected CNV. Single- and multi-marker (haplotype) statistical methods will be applied to the SNP data. De novo mutations and common copy number variation will be assessed. Also, novel statistical methods will be developed and applied to CNV data. Loci identified in this way will be ranked by significance. Replication of association for top-ranked SNP loci will be tested using similar statistical methods applied to genotypes obtained with the Illumina GoldenGate system in the 1,400 trios. Taqman and Sequenom-based platforms will be utilized to genotype the most promising, replicated loci in the third cohort of 1,000 trios. Further studies including in silico methods and direct DNA sequencing will be used to identify possible causal variants in disease-associated genes. These studies will reveal AIS susceptibility loci in genes with strong effects across patient populations. These findings will be important for many future studies ultimately targeting prevention or alternative treatments.