Discovery and Annotation of Targets for Gene Therapy of Infertile Men

翻译后摘要：项目一：不育男性基因治疗的目标的发现和注释 不孕症影响了美国15%的育龄夫妇，导致超过108，000例新的访问， 生殖内分泌学和不孕症诊所。几乎所有这些诊所都提供植入前 胚胎基因诊断和与内分泌功能障碍相关的已知突变的基因检测， 原发性性腺衰竭和反复流产然而，生殖医学专家依靠老年人 细胞遗传学、序列标记位点PCR和桑格测序等技术用于这些检测， 利用临床遗传学中常见的现代全基因组和RNA测序技术， 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癌症的方式。 治疗不孕症。此P50应用程序的总体目标是为如何使用这些工具创建路线图， (a)鉴定导致男性不育的突变，（B）表征这些突变如何导致 （c）基因疗法如何可以用于以安全和有效的方式治疗这些病理， 有针对性地在这个项目中，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麦基妇女医院。我们会把所有的- 基因组测序和高分辨率阵列CGH以绘制总共500个病例的遗传变异位置， 包括21个具有可遗传形式的无精子症的大的、多个家族。作为患者表型分析的一部分， 我们将部署一个专门的工具，称为查尔森科摩罗指数，专门记录 每例不孕症患者的合并症证据。我们将开发和应用高度敏感的统计数据， 方法，利用大量人口控制数据库中的信息， 可能导致生精障碍风险的异常突变。我们将发展知识 总结了模式生物的证据，表明这些突变可以导致两者的病理学 性腺和体细胞组织。我们将尝试推断睾丸细胞类型，这是主要的网站， 每个突变的病理学，以帮助指导基因治疗的靶向。 这项工作最重要的长期成果将是出版分析工具和知识 这将有助于基因组测序在不孕症治疗中的应用。结合结果 从项目II和项目III中，我们的结果将为生殖医学专家提供使用 现代遗传技术来研究和治疗不孕症。

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.