Functional dissection of GnRH defects and networks

项目3摘要 在越来越大的疾病队列中，扩大的基因分型和下一代测序已经显著 成功地识别了驱动病理和/或赋予易感性的新的基因座、基因和等位基因。 然而，这些努力遇到了三大障碍。首先，关联研究与一些值得注意的 由于难以有效地从定位基因座过渡到识别基因座，这些例子受到了阻碍 有贡献的基因和等位基因。其次，超罕见或私人的孟德尔突变一直在挑战 以足够的数量进行鉴定，以将它们与表型紧密联系起来。第三，压倒性的变异 在遗传异质性的促性腺激素释放激素(GnRH)疾病中被鉴定为非 等位基因致病性的同义词变化，仅用遗传论据很难推断。一 这些障碍的原因是人类中存在大量罕见的变异 人口和当前的生物信息学工具在准确预测这一比例方面仍然有限 变异是功能性的和表型相关的，与我们在项目1和2中的同事一起，我们有 交叉的遗传、基因组和功能工具，以克服其中的一些挑战，并已开始 告知生殖发育障碍的遗传结构。我们过去的协同努力导致了 RNF216和OTUD4基因是单纯性低促性腺激素减退症的首批突变基因 (IGD)在Gordon-Holmes综合征患者中建立功能障碍的“模块”，即 泛素-蛋白酶体途径在本病中的作用。作为这项工作的补充，系统在体内发挥功能 对CHD7的一个等位基因序列的评估使我们能够分离出该基因座对 GnRH轴在突变负荷假说下的生殖障碍，否则无法 经典的统计工具。在这些调查的下一阶段，项目3将扩大其作为桥梁的作用 在正在进行的和成功的孟德尔基因发现努力(项目1)和最先进的 复杂性状关联方法(项目2)。项目3将评估新候选人的致病性 基因和基因座分别来自项目2和项目3。它还将注释非同义的编码变体 在IGD患者和大量人群中发现作为这一过程的一部分，作为确定 受影响个体的突变负担。我们还将利用我们生成的体内模型来模拟 在患者队列中观察到的寡聚现象。同时，项目3将分离纯细胞种群 与斑马鱼模型中的GnRH生物学相关，以产生转录网络，从而告知 整体中心的研究。

Project 3 Abstract Expanded genotyping and next generation sequencing in ever-larger disease cohorts has been remarkably successful in identifying novel loci, genes and alleles that drive pathology and/or confer susceptibility. However, these efforts have encountered three major obstacles. First, association studies with some notable examples have been hampered by the difficulty in transitioning efficiently from mapping loci to identifying contributory genes and alleles. Second, ultra-rare or private Mendelian mutations have been challenging to identify in sufficient numbers to link them robustly to phenotype. Third, an overwhelming majority of variation identified in the genetically heterogeneous gonadotropin-releasing hormone (GnRH) disorders are non- synonymous changes for which allele pathogenicity is difficult to infer using genetic arguments alone. One reason for these impediments is the presence of the large amount of rare variation that exists in human populations and the current bioinformatic tools remain limited in predicting with accuracy which fraction of this variation is functional and phenotypically relevant, Together with our colleagues in Projects 1 and 2, we have intersected genetic, genomic and functional tools to overcome some of these challenges and have begun to inform the genetic architecture of disorders of reproductive development. Our past synergistic efforts led to the identification of RNF216 and OTUD4 as the first genes mutated in isolated hypogonadotropic hypogonadism (IGD) in individuals with Gordon-Holmes syndrome establishing a “module” of dysfunction, namely the ubiquitin-proteasome pathway in this disorder. Complementary to this work, systematic in vivo functional assessment of an allelic series in CHD7 allowed us to isolate the non-Mendelian contribution of this locus to reproductive disorders of the GnRH axis under a mutational burden hypothesis that was otherwise refractory to classical statistical tools. In the next phase of these investigations, Project 3 will expand its role as a bridge between the ongoing and successful Mendelian gene discovery efforts (Project 1) and the state-of-the-art complex trait association approaches (Project 2). Project 3 will assess the pathogenicity of novel candidate genes and loci from Projects 2 and 3 respectively. It will also annotate non-synonymous coding variants discovered as part of this process in IGD patients and large populations as a means of determining the mutational burden in affected individuals. We will also utilize the in vivo models we generate to model oligogenic phenomena observed in patient cohorts. In parallel, Project 3 will isolate pure cellular populations relevant to GnRH biology from zebrafish models to generate transcriptional networks that will inform the studies of the overall Center.