Familial idiopathic scoliosis: gene discovery and functional studies

 描述（由申请人提供）：家族性特发性脊柱侧凸：基因发现和功能研究特发性脊柱侧凸（IS）是一种病因不明的脊柱结构性侧弯。IS影响3%的其他正常儿童，其中女性严重进展的风险最大。目前的治疗选择仅限于观察，物理治疗，支具和手术，主要是针对解决身体和美容畸形。IS临床上以散发性和家族性两种形式发生，但目前对该疾病的遗传学和分子病因学均不清楚。IS基因和途径的发现将大大提高对这种疾病发病机制的整体理解，并有助于开发靶向诊断和治疗方法。IS的遗传学研究已经确定了可能导致该疾病的多个区域;然而，很少有基因直接参与IS的发病机制。从历史上看，在全基因组水平上筛选具有大个体效应的突变体的能力在经济上是不允许的。下一代（NextGen）DNA测序技术的进步允许经济的短读段DNA测序，专注于基因组（外显子组）的蛋白质编码方面。我们将把NextGen技术带到我们独特的家族性特发性脊柱侧凸（FIS）研究队列中，该队列包括725个家庭，2505名个体。我们检测与FIS病因相关的变异的创新策略是关注具有高疾病负担的家庭，因此， 可能隐藏着具有巨大影响的罕见变异。该提议的中心假设是，多种罕见的致病性编码变体负责FIS表型，并且将通过细胞和小鼠模型中的外显子组测序和功能实验来鉴定。我们的具体目标是：1）对27个多代同堂的大家庭进行外显子组测序，发现新的FIS基因（发现队列），2）在200个FIS家族的第二群体中验证这些新鉴定的FIS基因（验证组群），和3）研究细胞培养和小鼠模型系统中鉴定的变体的功能，旨在验证人类特异性FIS变异体和FIS病因学的机制性见解。我们的小组已经投入了20多年的研究和收集受严重FIS影响的大家庭。这一建议是一个新的方法，针对FIS基因的发现。这项工作的主要目标是确定有助于FIS表型的基因，这将为调查疾病易感性和严重畸形进展的机制提供切入点。这项工作的发现最终将使我们深入了解影响未成熟骨骼发育的生物学过程，并有助于推进FIS的个性化治疗干预。

 DESCRIPTION (provided by applicant): Familial idiopathic scoliosis: gene discovery and functional studies Idiopathic scoliosis (IS) is a structural lateral curvature of the spine of =10 of unknown etiology. IS affects 3% of otherwise normal children, with females at the greatest risk for severe progression. Current treatment options are limited to observation, physical therapy, bracing, and surgery, and are primarily directed to addressing the physical and cosmetic deformity. IS occurs clinically in both sporadic and familial forms, but neither the genetic nor th molecular etiology for the disorder is currently understood. The discovery of IS genes and pathways would greatly enhance the overall understanding of the pathogenesis of this disorder and aid in the development of targeted diagnostics and therapeutics. Genetic studies in IS have identified multiple regions that potentially contribute to the disorder; however, very few genes have been directly implicated in the pathogenesis of IS. Historically, the ability to screen for rae variants with large individual effects at the genome-wide level has been economically prohibitive. Advances in Next Generation (NextGen) DNA sequencing technology allow for economical short-read DNA sequencing, focusing on the protein coding aspects of the genome (the exome). We will bring NextGen technology to our unique familial idiopathic scoliosis (FIS) study cohort of 725 families with 2505 individuals. Our innovative strategy for the detection of variants related to FIS etiology is to focus on families with a high disease burden, and, thus, are likely to harbor rare variants with large effects. The central hypothesis of this proposal is that multiple rare pathogenic coding variants are responsible for the FIS phenotype and will be identified by exome sequencing and functional experiments in cells and mouse models. Our specific aims are: 1) to perform exome sequencing in 27 extended multigenerational families to discover new FIS genes (discovery cohort), 2) to validate these newly identified FIS genes in a second population of 200 FIS families (validation cohort), and 3) to investigate the function of identified variants in both cell culture and mouse model systems, directed to validation of human- specific FIS variants and mechanistic insight into the etiology of FIS. Our group has invested over 20 years studying and collecting large families affected by severe FIS. This proposal is a novel approach directed to FIS gene discovery. The primary goal of this work is to identify genes that contribute to the FIS phenotype, which will provide entry points for the investigation of the mechanisms underlying disease susceptibility and severe progression of deformity. The discoveries from this work will ultimately give us insight into the biological processes that influence the development of the immature skeleton and could aid in the advancement of personalized therapeutic interventions for FIS.