Functional dissection of GnRH defects and networks

GnRH 缺陷和网络的功能剖析

• 批准号: 9910434
• 负责人: Erica Ellen Davis
• 金额: $ 23.81万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2021-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9910434
• 关键词: Affect; Alleles; Area; Biological Assay; Biological Process; Biology; CHD7 gene; CRISPR/Cas technology; Candidate Disease Gene; Code; Collaborations; Complex; Defect; Development; Disease; Disease susceptibility; Dissection; Embryo; Endocrine; Endocrine System Diseases; Family; Functional disorder; GNRH2 gene; Gene Mutation; Generations; Genes; Genetic; Genetic Diseases; Genetic Epistasis; Genetic Transcription; Genetic Translation; Genetic study; Genomics; Genotype; Gonadotropin Hormone Releasing Hormone; Human; Idiopathic Hypogonadotropic Hypogonadism; Individual; Investigation; KISS1 gene; Lesion; Link; Maps; Modeling; Mutate; Mutation; Nature; Neurons; Outcome; Pathogenicity; Pathology; Pathway Analysis; Pathway interactions; Patients; Phase; Phenotype; Physiological; Population; Population Study; Predisposition; Privatization; Process; Refractory; Reproduction; Risk; Role; Science; Series; Speed; Syndrome; Testing; Ubiquitin; Variant; Work; Zebrafish; bioinformatics tool; cohort; exome sequencing; gene discovery; genetic architecture; genetic manipulation; genome sequencing; genomic locus; human model; in vivo; in vivo Model; insight; loss of function; migration; multicatalytic endopeptidase complex; mutant; next generation sequencing; novel; protein function; reproductive; reproductive development; tool; trait; transcriptome sequencing; transcriptomics; whole genome

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.

项目3 在越来越大的疾病队列中扩大基因分型和下一代测序已经显着 成功鉴定驱动病理和/或赋予易感性的新基因座、基因和等位基因。 然而，这些努力遇到了三大障碍。首先，与一些著名的 由于从定位基因座到识别 贡献基因和等位基因。其次，超罕见或私人孟德尔突变一直具有挑战性， 鉴定出足够数量以将它们与表型可靠地联系起来。第三，绝大多数变异 在遗传异质性促性腺激素释放激素（GnRH）疾病中发现的非- 同义变化，其等位基因致病性难以单独使用遗传参数推断。一 这些障碍的原因是存在于人类中的大量罕见变异， 人口和目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，我们与项目1和2的同事一起， 遗传学、基因组学和功能性工具来克服其中一些挑战，并已开始 为生殖发育障碍的遗传结构提供信息。我们过去的协同努力导致了 RNF 216和OTUD 4作为孤立性低促性腺激素性性腺功能减退症中第一个突变基因的鉴定 (IGD)在Gordon-Holmes综合征患者中建立功能障碍的“模块”，即 泛素-蛋白酶体途径在这种疾病中的作用。补充这项工作，系统的体内功能 对CHD 7中等位基因系列的评估使我们能够分离出该基因座的非孟德尔贡献， 根据突变负荷假设，GnRH轴的生殖障碍，否则难以治疗 经典统计工具。在这些调查的下一阶段，项目3将扩大其桥梁作用 正在进行的和成功的孟德尔基因发现工作（项目1）和最先进的 复杂性状关联方法（项目2）。项目3将评估新候选药物的致病性 基因和位点分别来自项目2和3。它还将注释非同义编码变体 在IGD患者和大量人群中发现，作为这一过程的一部分， 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建模 在患者队列中观察到寡基因现象。与此同时，项目3将分离纯细胞群体， 相关的GnRH生物学从斑马鱼模型，以产生转录网络，将告知 整个中心的研究。