Project 1 - Deciphering the Molecular Drivers of Rare Forms of Human Infertility Using Integrative Genomic, Cellular, and Phenomic Approaches

项目 1 - 使用综合基因组、细胞和表型组方法破译罕见人类不孕症的分子驱动因素

• 批准号: 10613359
• 负责人: Stephanie Beth Seminara
• 金额: $ 40.1万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-10 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10613359
• 关键词: Address; Affect; Alternative Splicing; Biological; Biological Models; CRISPR/Cas technology; Clinical; Clinical Investigator; Clustered Regularly Interspaced Short Palindromic Repeats; Code; Couples; Data; Development; Diagnosis; Diagnostic; Disease; Drosophila genus; Engineering; Evaluation; Fertility; GNRH1 gene; Gene Expression; General Hospitals; Genes; Genetic Counseling; Genetic Heterogeneity; Genetic Risk; Genetic Variation; Genomics; Genotype; Goals; Gynecologic; Health; Hospitals; Human; Human Genetics; Hypogonadism; Hypothalamic structure; Idiopathic Hypogonadotropic Hypogonadism; Individual; Induced pluripotent stem cell derived neurons; Infertility; Infertility study; KISS1 gene; Kallmann Syndrome; Massachusetts; Measures; Medical Genetics; Methodology; Modeling; Molecular; Morphology; Mutation; National Institute of Child Health and Human Development; Neurons; Ovarian; Ovary; Pathogenicity; Pathway interactions; Patients; Pattern; Penetrance; Phenotype; Physiologic pulse; Pituitary Gland; Population; Prevention strategy; Puberty; RNA Interference; Rare Diseases; Regulatory Pathway; Relative Risks; Reproductive Health; Reproductive Medicine; Research Personnel; Running; Saudi Arabia; Services; Single Nucleotide Polymorphism; Strategic Planning; Techniques; Technology; Universities; Untranslated RNA; Utah; Variant; biobank; causal variant; clinical care; cohort; diagnostic tool; exome; experimental study; genetic architecture; genetic variant; genome sequencing; human genomics; insertion/deletion mutation; knock-down; lens; migration; novel; novel diagnostics; novel therapeutics; overexpression; phenomics; precision medicine; premature; primary ovarian insufficiency; reproductive; reproductive function; reproductive hormone; risk prediction; risk variant; targeted treatment; transcriptome; transcriptomics; treatment strategy; variant detection; whole genome

PROJECT ABSTRACT Infertility affects up to 13% of childless couples yet the biological mechanisms underlying infertility and the effects of infertility on overall health remains poorly understood. The goal of Project 1 within The Massachusetts General Hospital Harvard Center for Reproductive Medicine will be to apply human genomics, transcriptomics and phenomics to understand the mechanistic biological drivers of rare disorders of human infertility. A pervasive methodologic theme of 'omics’ technologies runs through Project 1’s three specific aims. In Aim 1, infertility will be viewed through the prism of rare hypogonadotropic and hypergonadotropic conditions that cause infertility. Clinical investigators will apply contemporary genomic techniques to define the underlying genetic architecture of infertility and to identify master regulatory pathways and networks that determine fertility. Three unique clinical cohorts will be utilized to achieve this aim: US-based admixed cohorts of patients with idiopathic hypogonadotropic hypogonadism (IHH)/Kallmann Syndrome [Massachusetts General Hospital] and primary ovarian insufficiency [University of Utah]; and, a Saudi Arabia-based consanguineous cohort of patients with a spectrum of rare Mendelian forms of infertility. The full spectrum of genetic variation (coding and non-coding single nucleotide variants, insertion/deletion variants and structural variants) that confer substantial relative risk for infertility will be determined and causal genes identified will be coalesced into common, final pathways elucidating the predominant drivers of rare forms of infertility. In Aim 2, genetic variants identified from Aim 1 and variants identified in Project 2 of the Center that relate to hypothalamic-hypogonadotropic forms of infertility will be validated in CRISPR-engineered GnRH neurons derived from induced pluripotent stem cells generated by the Genomics & Functional Core of the Center. Specifically, the cellular and molecular consequences of genetic variation on GnRH neurons will be defined by comparing and contrasting the GnRH transcriptome, morphology, migratory capability and secretory function between wild-type and edited GnRH neurons. Similarly, genetic variants relating to hypergonadotropic forms of infertility leading to primary ovarian insufficiency will be studied in a Drosophila model system by ovary-specific RNAi knockdown (or overexpression) experiments. Finally, in Aim 3, a hospital-based human biobank (Partners Biobank) will be utilized to perform a recall-by-genotype based targeted phenotypic evaluation in individuals harboring pathogenic variants in infertility-associated genes. The full reproductive phenotype (“reproductome”) will be defined using deep phenotyping studies that will define the effects of harboring genetic risk variants on GnRH-induced pituitary LH pulse profiles and hypothalamic-pituitary responsiveness to exogenous kisspeptin administration. Through these coordinated studies, this project will aid in the diagnosis and management of infertility, inform genetic risk prediction for infertility conditions, and facilitate the development of novel therapeutic options for infertility.

项目摘要 不孕症影响高达 13% 的无子女夫妇，但不孕症背后的生物学机制及其影响 不孕不育对整体健康的影响仍然知之甚少。马萨诸塞州总医院项目 1 的目标 哈佛医院生殖医学中心将应用人类基因组学、转录组学和 表型组学来了解人类不孕症罕见疾病的生物学驱动因素。一种普遍存在的 “组学”技术的方法论主题贯穿项目 1 的三个具体目标。在目标 1 中，不孕症将 通过导致不孕的罕见促性腺激素低下和促性腺激素亢进病症的棱镜来观察。 临床研究人员将应用当代基因组技术来定义潜在的遗传结构 不孕症的研究并确定决定生育能力的主要调控途径和网络。临床三大独特 将利用队列来实现这一目标：美国特发性患者的混合队列 低促性腺激素性性腺功能减退症 (IHH)/卡尔曼综合征 [马萨诸塞州总医院] 和原发性 卵巢功能不全[犹他大学]；沙特阿拉伯的一个近亲结婚患者队列 罕见的孟德尔不孕不育症谱系。全谱遗传变异（编码和非编码） 单核苷酸变异、插入/缺失变异和结构变异）会带来相当大的相对风险 将确定不孕症的病因，并将确定的致病基因合并成共同的最终途径 阐明罕见形式不孕症的主要驱动因素。在目标 2 中，从目标 1 和 该中心项目 2 中发现的与下丘脑-促性腺激素分泌不足形式的不孕症相关的变异将 在源自诱导多能干细胞的 CRISPR 工程 GnRH 神经元中得到验证 该中心的基因组学和功能核心。具体来说，遗传的细胞和分子后果 GnRH 神经元的变异将通过比较和对比 GnRH 转录组、形态学、 野生型和编辑的 GnRH 神经元之间的迁移能力和分泌功能。同样，遗传 将研究与导致原发性卵巢功能不全的高促性腺激素不孕症相关的变异 在果蝇模型系统中，通过卵巢特异性 RNAi 敲低（或过度表达）实验。最后，在 目标 3，基于医院的人类生物库（合作伙伴生物库）将用于执行基于基因型的召回 对不孕相关基因携带致病性变异的个体进行有针对性的表型评估。这 完整的生殖表型（“生殖组”）将使用深度表型研究来定义，该研究将定义 携带遗传风险变异对 GnRH 诱导的垂体 LH 脉冲谱和下丘脑-垂体的影响 对外源性 Kisspeptin 给药的反应。通过这些协调研究，该项目将帮助 在不孕不育的诊断和管理中，为不孕不育的遗传风险预测提供信息，并促进 开发新的不孕不育治疗方案。