Transposons in Female Reproductive Health and Disease

描述（由申请人提供）：该提案描述了一个为期五年的培训计划，目的是作为NIH资助的生殖医学和不孕症独立调查员建立职业生涯。威廉姆斯博士对生殖生物学和妇女健康有着长期的兴趣和承诺，并在分子生物学和生殖医学方面有着良好的记录。他在Paul Wassarman博士的实验室获得了分子生物学博士学位，在那里他研究了哺乳动物卵母细胞。他在布里格姆妇女医院/马萨诸塞州总医院完成了妇产科住院医师培训，并在威尔康奈尔医疗中心接受了生殖内分泌学和不孕症研究金培训的最后一年。在这个奖项期间，他将扩大他在基因组学，RNA生物学，分析设计和计算生物学方面的能力，其总体目标是阐明转座子对卵母细胞和生育力的影响。 该培训计划旨在确保RNA和转座子生物学应用于女性生殖的命令。RNA生物学领域的先驱托马斯图舍尔博士将指导首席研究员的科学发展。他是洛克菲勒大学的生物化学教授，并培养和指导了许多博士后研究员独立的研究职位。培训活动将包括分析设计指导和RNA分子生物学和计算生物学讲座。一个由杰出的基础和医生科学家组成的科学咨询委员会将提供持续的反馈。 转座子是一种移动的遗传元件，占人类基因组的近一半。它们的复制会导致遗传损伤和疾病，必须在卵母细胞中抑制。在苍蝇和蠕虫中，一类特殊的小RNA分子（皮尔纳）和蛋白质（PIWI）抑制转座子。该研究的长期目标是研究哺乳动物卵母细胞抑制转座子的机制以及转座子对雌性生育力的影响。核心假设是转座子在哺乳动物中的复制可导致流产，并且通常被皮尔纳和PIWI蛋白抑制。将通过追求三个具体目标来测试该假设，即：（1）确定人卵巢是否含有皮尔纳和PIWI蛋白（2）评估雌性小鼠中PIWI敲除的表型（3）确定正常妊娠和流产中转座子复制的速率。该方法是创新的，因为它提出了不孕症的新病因，并涉及开发利用全基因组分析和第二代DNA测序的测定。这项研究意义重大，因为它将解释卵母细胞如何保护其基因组免受转座子的侵害，并为生育诊断和治疗提供新的途径。Tuschl博士在洛克菲勒大学的实验室是一个理想的环境，可以进行一系列高度专业化的科学技能的强化培训，以解决生殖生物学这个复杂的领域，并为生殖生物学的学术生涯做好主要研究员的准备。 公共卫生相关性：近一半的人类基因组由称为转座子的移动的遗传元件组成。卵母细胞必须抑制转座子复制，因为这会对基因组造成损害，可能导致流产、不育和早产。本研究的目的是确定卵母细胞如何抑制转座子以及转座子对雌性生殖的影响。

DESCRIPTION (provided by applicant): The proposal describes a five-year training program with the purpose of establishing a career as an NIH funded independent investigator in reproductive medicine and infertility. Dr. Williams has a longstanding interest in and commitment to reproductive biology and women's health and has a strong track record in molecular biology and reproductive medicine. He earned his PhD in Molecular Biology in the laboratory of Dr. Paul Wassarman, where he studied the mammalian oocyte. He completed residency training in obstetrics and gynecology at the Brigham and Women's Hospital/Massachusetts General Hospital and is in the last year of fellowship training in reproductive endocrinology and infertility at Weill-Cornell Medical Center. Over the course of this award period he will expand his proficiency in genomics, RNA biology, assay design and computational biology with the overarching goal of elucidating the effect of transposons on oocytes and fertility. The training program has been designed to ensure command of RNA and transposon biology as applied to female reproduction. Dr. Thomas Tuschl, a pioneer in the field of RNA biology, will mentor the principal investigator's scientific development. He is Professor of Biochemistry at The Rockefeller University and has trained and guided many postdoctoral fellows to independent research positions. Training activities will include instruction in assay design and lectures in RNA molecular biology and computational biology. A scientific advisory committee of exceptional basic and physician-scientists will provide ongoing feedback. Transposons are mobile genetic elements comprising nearly half of the human genome. Their replication can cause genetic damage and disease and must be suppressed in the oocyte. In flies and worms, a special class of small RNA molecules (piRNA) and proteins (PIWI) suppress transposons. The long-term goal of the study is to investigate the mechanism by which mammalian oocytes suppress transposons and the effect of transposons on female fertility. The central hypothesis is that replication of transposons in mammals can cause miscarriage and is normally suppressed by piRNA and PIWI proteins. The hypothesis will be tested by pursuing three specific aims, namely: (1) to determine whether human ovaries contains piRNA and PIWI proteins (2) to assess the phenotype of the PIWI knockout in the female mouse (3) to determine the rate of transposon replication in normal pregnancies and miscarriage. The approach is innovative because it proposes a new etiology of infertility and involves developing assay utilizing whole-genome analysis with second- generation DNA sequencing. The research is significant because it will explain how the oocyte protects its genome from transposons and provides a new avenue for fertility diagnosis and treatment. Dr. Tuschl's laboratory at Rockefeller University is an ideal setting for intensive training in a set of highly specialized scientific skills required to address this complex area of reproductive biology and prepare the principal investigator for an academic career in reproductive biology. PUBLIC HEALTH RELEVANCE: Nearly half of the human genome is composed of mobile genetic elements called transposons. The oocyte must suppress transposon replication because that causes damage to the genome that may result in miscarriage, infertility and premature. The purpose of this study is to determine how the oocyte suppresses transposons and the effect of transposons on female reproduction.