Discovery and Annotation of Targets for Gene Therapy of Infertile Men

摘要：项目I：男性不育基因治疗靶点的发现和注释 在美国，15%的育龄夫妇会受到不孕不育的影响，导致超过10.8万名新人前往 生殖内分泌和不孕症(REI)诊所每年。几乎所有这样的诊所都提供植入前的服务。 胚胎的基因诊断和与内分泌功能障碍相关的已知突变的基因测试， 原发性腺功能衰竭和反复妊娠丢失。然而，生殖医学专家依赖于老年人 细胞遗传学、序列标记位点聚合酶链式反应和桑格测序等技术用于这些测试，而不是 利用现代全基因组和RNA测序技术在临床遗传学中的普遍应用 其他疾病状态。 新的基因组工具，无论是计算的还是实验的，都有望彻底改变我们诊断和 治疗不孕不育。这个P50应用程序的总体目标是为如何使用这些工具创建一个路线图 (A)确定导致男性不育的突变；(B)确定这些突变可能如何促成男性不育 以及(C)如何使用基因疗法在安全和安全的情况下治疗这些病理学。 有针对性的方式。在这个项目中，我们从三个主要地点招募男性不育患者： 华盛顿大学、威尔·康奈尔医学院和玛吉妇女医院。我们将适用整个- 基因组测序和高分辨率阵列CGH在500例总病例中绘制遗传变异位置 包括21个可遗传形式的无精症大家庭。作为患者表型鉴定的一部分， 我们将部署一种名为Charlson共病指数的专门工具来专门记录 每一种不孕不育病例共病的证据。我们将开发和应用高度敏感的统计学 方法利用海量人口控制数据库中的信息进行统计识别 可能导致生精障碍风险的不寻常突变。我们将发展知识 总结了来自模型生物的证据的基础，这些证据表明这些突变可以导致两种疾病的病理 性腺和体细胞组织。我们将尝试推断睾丸细胞的类型(S)，这些细胞是 针对每个突变进行病理检查，以帮助指导靶向基因治疗。 这项工作最重要的长期成果将是分析工具和知识的出版 将促进基因组测序在不孕不育治疗中使用的碱基。结合结果 在项目II和项目III中，我们的结果将为生殖医学专家提供使用 研究和治疗不孕不育症的现代基因技术。

Abstract: Project I: Discovery and Annotation of Targets for Gene Therapy of Infertile Men Infertility affects 15% of reproductive-age couples in the US, leading to more than 108,000 new visits to reproductive endocrinology and infertility (REI) clinics per year. Nearly all such clinics offer pre-implantation genetic diagnosis of embryos and genetic testing for known mutations associated with endocrine dysfunction, primary gonadal failure, and recurrent pregnancy loss. However, reproductive medicine specialists rely on old technologies like cytogenetics, sequence-tagged site PCR, and Sanger sequencing for these tests and are not taking advantage of modern whole-genome and RNA sequencing technologies common in clinical genetics of other disease states. New genomic tools, both computational and experimental, promise to revolutionize the way we diagnose and treat infertility. An overall goal of this P50 application is to create a roadmap for how these tools can be used to (a) identify mutations contributing to male infertility, (b) characterize how these mutations may contribute to pathology in somatic tissues, and (c) how gene therapy can be used to treat these pathologies in a safe and targeted manner. In this project, we are recruiting patients of male infertility from three primary sites: Washington University, Weill Cornell Medical School, and Magee-Womens Hospital. We will apply whole- genome sequencing and high-resolution array CGH to map the location of genetic variation in 500 total cases, including 21 large, multiplex families with heritable forms of azoospermia. As part of the patient phenotyping, we will deploy a specialized instrument known as the Charlson Comorbidity index to specifically document the evidence for comorbidity in each case of infertility. We will develop and apply highly sensitive statistical methods, which draw upon the information in massive population control databases, to identify statistically unusual mutations that are likely to confer risk for spermatogenic impairment. We will develop knowledge bases that summarize the evidence from model organisms that these mutations can cause pathology in both gonadal and somatic tissues. And we will attempt to infer the testicular cell type(s) that are the primary sites of pathology for each mutation, to help guide the targeting of gene therapy. The most important long-term outcome of this work will be the publication of analysis tools and knowledge bases that will facilitate the use of genome sequencing in the treatment of infertility. Combined with the results from Project II and Project III, our results will give reproductive medicine specialists a roadmap for the use of modern genetic technologies to investigate and treat infertility.