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，但不孕症的生物学机制及其影响 不孕症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 医院哈佛生殖医学中心将应用人类基因组学、转录组学和 表型组学旨在了解人类不孕症罕见疾病的生物学驱动因素。一个普遍 “组学”技术的方法论主题贯穿于项目1的三个具体目标。在目标1中，不孕症将 通过罕见的低促性腺激素和高促性腺激素的条件下，导致不孕症的棱镜。 临床研究人员将应用当代基因组技术来确定潜在的遗传结构 并确定决定生育能力的主要调控途径和网络。三个独特的临床 队列将用于实现这一目标：基于美国的特发性乳腺癌患者混合队列 低促性腺激素性腺功能减退症（IHH）/Kallmann综合征[马萨诸塞州总医院]和原发性 卵巢功能不全[犹他州大学];以及，一个基于沙特阿拉伯的近亲患者队列， 一系列罕见的孟德尔式不育症全谱遗传变异（编码和非编码 单核苷酸变异、插入/缺失变异和结构变异）， 不孕症的病因将被确定，已确定的致病基因将被整合到共同的最终途径中， 阐明了罕见不孕症的主要驱动因素。在目标2中，从目标1和目标2中鉴定的遗传变异体被用于基因治疗。 该中心项目2中确定的与下丘脑-低促性腺激素型不孕症相关的变异将 在CRISPR工程化的GnRH神经元中得到验证，这些神经元来源于通过以下方法产生的诱导多能干细胞： 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 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，基因 将研究与导致原发性卵巢功能不全的高促性腺激素性不孕症相关的变异 在果蝇模型系统中通过卵巢特异性RNAi敲低（或过表达）实验。最后在 目标3，将利用基于医院的人类生物库（Partners Biobank）进行基于基因型的召回 对携带不育相关基因致病变异体的个体进行有针对性的表型评价。的 将使用深入的表型研究来定义完整的生殖表型（“reproductome”）， 携带遗传风险变异体对GnRH诱导的垂体LH脉冲曲线和下丘脑-垂体 对外源性kisspeptin给药的反应性。通过这些协调的研究，该项目将有助于 在不孕症的诊断和管理中，为不孕症的遗传风险预测提供信息， 开发治疗不孕症的新选择。