Regulation of Leukemia Inhibitory Factor (LIF) for early embryo implantation

简介(申请人提供)：西尔维娅·拉莫斯是医学博士，在内科、自身免疫性疾病以及在生殖生物学和临床胚胎学的基础科学方面接受过扎实的培训，拥有丰富的临床经验。她致力于不明原因不孕症的研究。西尔维亚的长期职业目标是成为一名独立资助的科学家，从事生殖生物学和RNA新陈代谢的基础研究，有意义的目标是将其转化为对人类不孕不育的解释。Silvia的短期职业目标是1)获得关于RNA新陈代谢的更具体和深入的知识；2)掌握使用siRNA或shRNA击倒特定蛋白质的技术；3)培育基因工程小鼠以在体内测试模型；4)获得负责任的研究指导方面的额外培训；5)发展写作技能、资质、初步数据和出版物，以可信地成为R01的PI，并确立自己作为生殖和RNA领域的全国领导者的地位。建议的研究计划、职业发展活动、导师团队和机构环境都非常适合帮助申请者实现这些目标。本项目的目的是研究一种特定的植入因子--白血病抑制因子(LIF)的调节。寿命 是成功将胚胎植入小鼠子宫所必需的。我们的假设是白血病抑制因子(LIF)在转录后水平受到调控。在目标1中，PI选择性地过度表达和下调人类细胞系中的每一种ZFP36蛋白(TTP、ZFP36L1和Zfp36L2)，以确定这些蛋白是否能够破坏LIF mRNA的稳定。在目标2中，PI将鉴定和表征哪些LIF mRNA序列参与与ZFP36蛋白的结合。在目标3中，PI将通过建立一个在子宫上皮细胞中表达高于正常水平的ZFP 36蛋白的小鼠模型，来测试ZFP 36蛋白在植入过程中LIF mRNA失稳的生物学相关性。这项建议的预期结果是改善对特定植入因子LIF的调节机制的了解，以及这种调节如何影响早期胚胎植入的过程。为了支持拉莫斯博士的职业发展，她将继续攻读基础高级分子生物学、在老鼠身上建立人类疾病模型以及研究伦理学方面的课程。指导团队将指导拉莫斯博士的研究和职业发展，其中包括国际公认的、独立资助的研究人员，他们在基因调控和RNA新陈代谢(Marzluff)以及人类疾病基因工程小鼠的生成和植入生物学(Caron)方面具有专业知识。研究环境将为实现上述研究和培训目标提供一个富有成效的、合作的和合作的氛围。 公共卫生相关性：女性不孕不育是一个世界性问题，影响到15亿育龄妇女中的10%。在美国，这相当于210万年龄在15-44岁之间的已婚女性不育--对她们来说，15%的不孕原因仍然不明。在过去的二十年里，大多数发达国家一直存在推迟生育的趋势，这增加了与年龄有关的不孕症。在美国，有730万15-44岁的女性使用过不孕不育服务。植入人体的相对低效在很大程度上仍然令人费解。这项研究和后续研究的目的是研究白血病抑制因子(LIF)的转录后调控及其对体内植入的影响。因此，新产生的信息可能会为不明原因的女性不孕症带来新的治疗选择。

DESCRIPTION (provided by applicant): Silvia Ramos is an MD PhD with a diverse portfolio including a strong clinical formation in internal medicine, autoimmune disease as well as a solid training in basic science of reproductive biology and clinical embryology. She is committed to devoting herself to the study of infertility of unexplained basis. Silvia's long-term-career goal i to be an independent funded scientist, doing basic research on reproductive biology and RNA metabolism with a meaningful goal to translate into providing explanations of human infertility of unexplained basis. Silvia's short-term career goals are to 1) gain a more specific and in depth knowledge on RNA metabolism; 2) master the techniques of knocking down specific proteins either using siRNA or shRNA; 3) generate a genetic engineered mouse to test the model in vivo; 4) obtain additional training in responsible conduct of research; and 5) develop writing skills, grantsmanship, preliminary data and publications to be credible as the PI of an R01 and establish herself as a national leader in the reproductive and RNA field. The proposed research plan, career development activities, mentorship team, and institutional environment are all uniquely suited to assist the applicant in achieving these goals. The goal of this project is to investigate the regulation of a specific implantation factor, Leukemia Inhibitory Factor (LIF). LIF is required for successful embryonic implantation to the uterus in mice. Our hypothesis is that Leukemia Inhibitory Factor (LIF) is regulated at the post-transcriptional level. In aim 1, the PI wll selectively overexpress and down-regulate each of the ZFP36 proteins (TTP, ZFP36L1 and ZFP36L2) in human cell lines to determine whether LIF mRNA can be destabilized by these proteins. In Aim 2, the PI will identify and characterize which sequences of LIF mRNA are involved in binding to ZFP36 proteins. In Aim 3, the PI will test the biological relevance of LIF mRNA destabilization by ZFP36 proteins during the implantation by creating a mouse model expressing higher than normal levels of one ZFP36 protein in uterine epithelial cells. The expected outcome of this proposal is an improved knowledge of the mechanisms by which a specific implantation factor, LIF, is regulated; and how this regulation influences the process of early embryo implantation. To support Dr. Ramos's career development, she will pursue coursework in basic advanced molecular biology, modeling human disease in mice, and research ethics. The mentorship team, which includes internationally-recognized, independently-funded investigators with expertise in gene regulation and RNA metabolism (Marzluff); and generation of genetically engineered mice of human disease and biology of implantation (Caron), will guide Dr. Ramos's research and career development. The research environment will provide a productive, collegial, and collaborative atmosphere ideal for pursuing the above research and training goals. PUBLIC HEALTH RELEVANCE: Female infertility is a worldwide problem affecting 10% of the 1.5 billion women at reproductive age. In US this corresponds to 2.1 million married women ages 15-44 who are infertile-for whom the cause of infertility remains unknown for 15%. During the past two decades there has been a consistent trend towards delayed childbearing in most developed countries, increasing age-related infertility. In the US, 7.3 million women ages 15-44 have used infertility services. The relative inefficiency of implantation in humans remains, to a great extent, inexplicable. The goal of this and subsequent research is to investigate the post-transcriptional regulation of the Leukemia Inhibitory Factor (LIF) and its effect on implantation i vivo. New generated information may, consequently, bring novel treatment options for female infertility of unexplained basis.