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

描述（由申请人提供）：Silvia拉莫斯是一名医学博士，拥有多元化的投资组合，包括内科学、自身免疫性疾病的强大临床形成以及生殖生物学和临床胚胎学基础科学的扎实培训。她致力于不明原因不孕症的研究。Silvia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代谢的基础研究，其有意义的目标是为无法解释的人类不育症提供解释。Silvia的短期职业目标是：1）获得关于RNA代谢的更具体和深入的知识; 2）掌握使用siRNA或shRNA敲除特定蛋白质的技术; 3）产生基因工程小鼠以在体内测试模型; 4）获得负责任的研究行为的额外培训;和5）发展写作技巧，granische，初步数据和出版物是可信的作为PI的R 01和建立自己作为一个国家的领导人在生殖和RNA领域。拟议的研究计划，职业发展活动，导师团队和机构环境都是唯一适合帮助申请人实现这些目标。本项目的目的是研究白血病抑制因子（LIF）的调控。LIF 是小鼠胚胎成功植入子宫所必需的。我们的假设是白血病抑制因子（LIF）是在转录后水平调节。在目的1中，PI将在人细胞系中选择性地过表达和下调ZFP 36蛋白（TTP、ZFP 36 L1和ZFP 36 L2）中的每一种，以确定LIF mRNA是否可以被这些蛋白质去稳定。在目标2中，PI将鉴定并表征LIF mRNA的哪些序列参与与ZFP 36蛋白的结合。在目标3中，PI将通过创建子宫上皮细胞中表达高于正常水平的一种ZFP 36蛋白的小鼠模型，检测植入期间ZFP 36蛋白导致LIF mRNA不稳定的生物学相关性。这一建议的预期结果是一个特定的植入因子，LIF，调节的机制，以及这种调节如何影响早期胚胎植入过程的知识。为了支持拉莫斯博士的职业发展，她将继续学习基础先进分子生物学、小鼠人类疾病建模和研究伦理学等课程。导师团队，其中包括国际公认的，独立资助的研究人员在基因调控和RNA代谢（Marzluff）的专业知识;和人类疾病和植入生物学（卡隆）的基因工程小鼠的一代，将指导博士拉莫斯的研究和职业发展。研究环境将为追求上述研究和培训目标提供一个富有成效的，合议和协作的氛围。

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.