Dissecting ocular congenital cranial dysinnervation disorders through whole genome sequence analysis

项目概要（摘要） 这项建议的目的是确定新的遗传原因的眼睛`先天性颅神经支配障碍 疾病“（CCDD）的全基因组序列（WGS）数据的彻底分析，并定义表型- 这些新发现的CCDD疾病的基因型相关性和神经发育机制 基因.据估计，每1000名婴儿中就有1人出生时无法移动一只或两只眼睛。 在一个或多个方向。这种疾病会导致严重的残疾，并经常伴有 额外的结构性出生缺陷，通常在家庭内分离或由新生突变引起。MPI 恩格尔的遗传和发育研究导致这些综合征的定义，作为一个新的类别， 人类疾病她的实验室已经定义了多种CCDD综合征，揭示了它们的遗传病因， 通过模型生物的建模，确定这些疾病可能是由于发育不良， 颅运动神经元及其轴突突起。尽管取得了这些成功，超过80%的恩格尔实验室眼 CCDD队列的遗传学问题仍未解决。为了鉴定新的眼部CCDD基因，MPI Engle已被授予 来自基因未定义的CCDD先证者和家族成员的DNA样本的WGS，通过Gabriella 米勒儿童第一儿科研究计划。WGS的分析将允许检测非编码变体，拷贝 数字变化和复杂的结构重排，同时也提供了更好的编码覆盖面 区域，从而填补了其他遗传方法所遗漏的几个关键空白， 外显子组测序MPI麦克阿瑟是大型数据集基因组分析的国际领导者， 罕见疾病的背景。他的团队将分析来自700多名患有眼外伤的个人和家庭成员的WGS。 CCD，提供严格的数据处理，并与MPI Engle的团队密切合作，以评估证据 支持变异致病性。至关重要的是，MPI麦克阿瑟的参与将使我们能够分析我们的 在布罗德研究所测序的20，000多个对照基因组的背景下， 结构性出生缺陷基因组由儿童第一财团产生。结合靶向测序， Engle CCDD数据库中的其他先证者，这种协调将增强我们确定 致病性和表型-基因型相关性。使用MPI中建立的函数方法 Engle的实验室研究眼CCDD的神经发育和机制病因，高置信度 新的疾病基因将被转移到体外和体内的功能研究。因此，我们希望分析 这一独特的患者队列将导致对CCDD缺失的单基因病因的识别， 验证，复制和功能研究将阐明新的遗传和发育途径， 眼脑神经发育反过来，这将加强对患者的遗传诊断和咨询， 具有眼CCDD的家庭，告知健康和疾病中的运动神经元和轴突发育， 改善治疗方法，减少继发于这些孟德尔疾病的残疾。

PROJECT SUMMARY (ABSTRACT) The goals of this proposal are to identify novel genetic causes of ocular `congenital cranial dysinnervation disorders' (CCDDs) thorough analysis of whole genome sequence (WGS) data and to define the phenotype- genotype correlations and neurodevelopmental mechanisms underlying these newly identified CCDD disease genes. It is estimated that more than 1 of every 1000 infants is born with the inability to move one or both eyes in one or more directions. Such disorders cause significant disability and are frequently accompanied by additional structural birth defects, and often segregate within families or arise from de novo mutations. MPI Engle's genetic and developmental studies have led to the definition of these syndromes as a new category of human disease. Her lab has defined multiple CCDD syndromes, uncovered their genetic etiologies, and, through modeling in model organisms, determined that these disorders can result from maldevelopment of cranial motor neurons and their axonal processes. Despite these successes, over 80% of the Engle Lab ocular CCDD cohort remains genetically unsolved. To identify new ocular CCDD genes, MPI Engle has been granted WGS of DNA samples from genetically undefined CCDD probands and family members through the Gabriella Miller Kids First Pediatric Research Program. Analysis of WGS will allow detection of non-coding variants, copy number variations, and complex structural rearrangements, while also providing better coverage of coding regions than exome sequencing, thus filling several critical gaps missed by other genetic approaches such as exome sequencing. MPI MacArthur is an international leader in the genomic analysis of large datasets in the context of rare disease. His team will analyze the WGS from >700 individuals and family members with ocular CCDDs, provide rigorous data processing, and work closely with MPI Engle's team to evaluate evidence supporting variant pathogenicity. Critically, the involvement of MPI MacArthur will allow us to analyze our samples in the context of over 20,000 control genomes sequenced at the Broad Institute, as well as additional structural birth defect genomes generated by the Kids First consortium. Together with targeted sequencing of additional probands in the Engle CCDD database, this harmonization will enhance our power to determine pathogenicity and phenotype-genotype correlations. Employing the functional approaches established in MPI Engle's lab to study the neurodevelopmental and mechanistic etiologies of ocular CCDDs, high-confidence novel disease genes will be moved to functional studies in vitro and in vivo. Thus, we expect that analysis of this unique patient cohort will lead to the identification of missing monogenic causes of CCDDs and that validation, replication, and functional studies will elucidate new genetic and developmental pathways critical to ocular cranial nerve development. In turn, this will enhance genetic diagnoses and counseling in patients and families with ocular CCDDs, inform motor neuron and axon development in health and disease, and contribute to improved therapies and reduced disabilities that arise secondary to these Mendelian disorders.