Transposons in Female Reproductive Health and Disease

描述(由申请者提供)：该提案描述了一个为期五年的培训计划，目的是建立一个由NIH资助的生殖医学和不孕不育的独立研究人员的职业生涯。威廉姆斯博士长期关注并致力于生殖生物学和妇女健康，在分子生物学和生殖医学方面有着良好的记录。他在保罗·瓦萨曼博士的实验室获得了分子生物学博士学位，在那里他研究哺乳动物的卵母细胞。他在布里根妇女医院/马萨诸塞州综合医院完成了产科和妇科住院医师培训，并在威尔-康奈尔医学中心接受了生殖内分泌学和不孕不育方面的最后一年团契培训。在这一获奖期间，他将扩展他在基因组学、RNA生物学、分析设计和计算生物学方面的熟练程度，首要目标是阐明转座子对卵母细胞和生育能力的影响。该培训计划旨在确保掌握RNA和转座子生物学，并将其应用于雌性生殖。RNA生物学领域的先驱Thomas Tuschl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，曾培训和指导许多博士后研究员担任独立研究职位。培训活动将包括测试设计的指导以及RNA分子生物学和计算生物学的讲座。一个由杰出的基础科学家和内科科学家组成的科学咨询委员会将提供持续的反馈。转座子是一种可移动的遗传元件，占人类基因组的近一半。它们的复制会导致基因损伤和疾病，必须在卵母细胞中被抑制。在苍蝇和蠕虫中，一类特殊的小RNA分子(PiRNA)和蛋白质(PIWI)抑制转座子。这项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能力的影响。中心假说是，在哺乳动物中复制转座子会导致流产，通常会被piRNA和PIWI蛋白抑制。这一假设将通过追求三个具体目标来检验，即：(1)确定人类卵巢是否含有PiRNA和PIWI蛋白(2)评估雌性小鼠PIWI基因敲除的表型(3)确定正常妊娠和流产中转座子复制率。该方法具有创新性，因为它提出了一种新的不孕症病因，并涉及利用全基因组分析和第二代DNA测序开发检测方法。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不受转座子的影响，并为生育诊断和治疗提供一条新的途径。图施尔博士在洛克菲勒大学的实验室是一系列高度专业化的科学技能强化培训的理想场所，这些技能需要解决生殖生物学这一复杂领域的问题，并为生殖生物学的学术生涯做好主要研究人员的准备。 与公共卫生相关：近一半的人类基因组由称为转座子的可移动遗传元件组成。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，从而可能导致流产、不孕和早产。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.