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

白血病抑制因子（LIF）对早期胚胎植入的调节

• 批准号: 8383874
• 负责人: Silvia Beatriz Ramos
• 金额: $ 11.31万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8383874
• 关键词: 3' Untranslated Regions; Address; Adenine; Affect; Age; Autoimmune Diseases; Basic Science; Binding; Biological; Biological Testing; Cell Line; Cells; Child; Clinical; Commit; Couples; Cytoplasm; Data; Delayed Childbearing; Developed Countries; Doctor of Philosophy; Electrophoretic Mobility Shift Assay; Elements; Embryo; Embryology; Endometrium; Environment; Epithelial Cells; Epithelium; Estrus; Family; Family member; Female infertility; Funding; Gene Expression Regulation; Generations; Genetic Engineering; Genetic Transcription; Genetically Engineered Mouse; Goals; Human; Human Biology; Human Cell Line; Immunologic Adjuvants; Infertility; Infertility study; Internal Medicine; Investigation; Kinetics; Knowledge; Lead; Luciferases; Mentorship; Messenger RNA; Metabolism; Modeling; Molecular; Molecular Biology; Mus; Outcome; Peptides; Physiological; Poly I-C; Post-Transcriptional Regulation; Pregnancy; Process; Progesterone Receptors; Protein Overexpression; Proteins; Publications; RNA; RNA Decay; RNA Degradation; Regulation; Relative (related person); Reporter; Reproductive Biology; Research; Research Ethics; Research Personnel; Research Training; Role; Scientist; Services; Small Interfering RNA; Solid; Spontaneous abortion; System; TIS11 protein; Techniques; Testing; Training; Transcript; Transcription Process; Transgenic Mice; Translating; Uridine; Uterus; Woman; Writing; ZFP36L2 gene; Zinc Fingers; age related; base; career; career development; clinical practice; global health; human disease; implantation; improved; in vivo; in vivo Model; knock-down; leukemia inhibitory factor; mRNA Transcript Degradation; member; mouse model; mutant; natural Blastocyst Implantation; novel; overexpression; planetary Atmosphere; promoter; reproductive; research and development; responsible research conduct; skills; small hairpin RNA; spatiotemporal; trend

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.

描述（由申请人提供）：西尔维娅·拉莫斯（Silvia Ramos）是一名医学博士，拥有多元化的专业知识，包括内科、自身免疫性疾病方面的强大临床知识以及生殖生物学和临床胚胎学基础科学方面的扎实培训。她致力于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人类无法解释的不孕症提供解释。 Silvia 的短期职业目标是 1) 获得有关 RNA 代谢的更具体和更深入的知识； 2）掌握使用siRNA或shRNA敲低特定蛋白质的技术； 3）生成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； 5) 培养写作技巧、资助能力、初步数据和出版物，使其成为 R01 的 PI 的可信者，并将自己确立为生殖和 RNA 领域的国家领导者。拟议的研究计划、职业发展活动、导师团队和机构环境都非常适合帮助申请人实现这些目标。该项目的目标是研究特定植入因子——白血病抑制因子（LIF）的调节。利夫 是胚胎成功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节。在目标 1 中，PI 将选择性过表达和下调人类细胞系中的每种 ZFP36 蛋白（TTP、ZFP36L1 和 ZFP36L2），以确定 LIF mRNA 是否会被这些蛋白破坏稳定。在目标 2 中，PI 将识别并表征哪些 LIF mRNA 序列参与与 ZFP36 蛋白的结合。在目标 3 中，PI 将通过创建一个在子宫上皮细胞中表达高于正常水平的一种 ZFP36 蛋白的小鼠模型，来测试植入过程中 ZFP36 蛋白导致 LIF mRNA 不稳定的生物学相关性。该提案的预期结果是提高对特定植入因子 LIF 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，她将学习基础高级分子生物学、小鼠人类疾病建模和研究伦理学课程。导师团队包括国际公认的、独立资助的研究人员，他们在基因调控和 RNA 代谢方面拥有专业知识 (Marzluff)；人类疾病和植入生物学的基因工程小鼠的产生（Caron），将指导拉莫斯博士的研究和职业发展。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、学术和协作的氛围。 公共卫生相关性：女性不孕症是一个世界性问题，影响着 15 亿育龄妇女中的 10%。在美国，这相当于 210 万年龄在 15 岁至 44 岁之间的已婚女性不孕——其中 15% 的不孕原因仍不清楚。在过去的二十年里，大多数发达国家一直存在延迟生育的趋势，导致与年龄相关的不孕症增加。在美国，有 730 万 15-44 岁的女性使用过不孕不育服务。人体植入的相对低效率在很大程度上仍然令人费解。本研究及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，新生成的信息可能为不明原因的女性不孕症带来新的治疗选择。