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 不育影响多达13％的无子女夫妇，但不育的生物学机制和影响 对整体健康的不孕症仍然知之甚少。马萨诸塞州一般项目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 了解人类不育症罕见疾病的机械生物学驱动因素。普遍 “ Omics”技术的方法论主题贯穿项目1的三个特定目标。在AIM 1中，不育会 可以通过罕见的降压和高单核疾病的棱镜来看待。 临床研究人员将应用现代基因组技术来定义潜在的遗传结构 不孕症并确定确定生育能力的主要监管途径和网络。三个独特的临床 将利用队列来实现这一目标：基于美国的特发性患者的混合物队列 性腺功能低下（IHH）/Kallmann综合征[马萨诸塞州综合医院]和初级 卵巢不足[犹他大学]；而且，由沙特阿拉伯基于沙特阿拉伯的血统队列 遗传变异（编码和非编码）的全光谱 单个核苷酸变体，插入/缺失变体和结构变体）会遇到实质性相对风险 对于不孕症，确定的因果基因将合并为通用的最终途径 阐明罕见形式不育的主要驱动因素。在AIM 2中，从AIM 1和 中心项目2中确定的变体与下丘脑 - 高核核形式的不育形式将 在CRISPR设计的GnRH神经元中得到验证，该神经元来自于诱导的多能干细胞产生的诱导多能干细胞 中心的基因组学和功能核心。具体而言，通用性的细胞和分子后果 GNRH神经元的变化将通过比较和对比GNRH转录组，形态，， 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，通用 与不育的高核形式有关的变体将研究导致原发性卵巢不足 在果蝇模型系统中，通过卵巢特异性RNAi敲低（或过表达）实验。最后，在 AIM 3，将利用医院的人类生物库（合作伙伴生物库）进行基于偶然的召回型 在与不育相关基因中具有致病变异的个体中的靶向表型评估。这 将使用深层表型研究来定义完整的生殖表型（“生殖器”） 携带遗传风险变异对GNRH诱导的垂体LH脉冲曲线和下丘脑垂体的影响 对外源性亲吻肽给药的反应。通过这些协调的研究，该项目将有助于 在不育的诊断和管理中 不孕症的新型治疗选择的发展。