MOLECULAR AND METABOLIC ASPECTS OF IMPLANTATION

植入的分子和代谢方面

• 批准号: 9173780
• 负责人: KELLE H MOLEY
• 金额: $ 37.87万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9173780
• 关键词: Address; Affect; Autophagocytosis; Biochemical; Biological Assay; Cells; Clinical Trials; Complication; Data; Decidual Cell Reactions; Decidualization failure; Defect; Development; Diet; Drops; Embryo; Endometrial Stromal Cell; Epithelial; Event; Exposure to; Fertility; Funding; Genes; Genetic; Glucose; Glucose Transporter; Glycolysis; Goals; Grant; Guidelines; Hepatocyte; High Fat Diet; Hormones; Human; In Vitro; Knock-out; Knockout Mice; Knowledge; Measures; Metabolic; Molecular; Mus; NADP; Nonesterified Fatty Acids; Nucleotides; Obesity; Outcome; Palmitic Acids; Pathway interactions; Pentosephosphate Pathway; Play; Pregnancy; Pregnancy Outcome; Pregnancy Rate; Pregnancy loss; Process; Production; Progesterone; Proliferating; Recycling; Regulation; Ribose; Risk; Role; SLC2A1 gene; SLC2A12 gene; Saturated Fatty Acids; Signal Pathway; Staging; Stromal Cells; Testing; Woman; Work; blastocyst; blastomere structure; effective therapy; failure Implantation; implantation; improved; in vivo; inhibition of autophagy; inorganic phosphate; mouse model; natural Blastocyst Implantation; novel therapeutic intervention; novel therapeutics; pre-clinical; preimplantation; prevent; reproductive; research study; trophoblast

PROJECT SUMMARY Early pregnancy loss affects roughly 15% of known pregnancies and may be even more common in obese women. A prominent cause of early pregnancy loss is failure of implantation, which requires that the uterine endometrial stromal cells undergo decidualization. Additionally, recent work has revealed an important role for metabolic regulation in in this process. Specifically, the decidualizing stromal cells produced less ATP and more NADPH and ribose-5-phosphate via the pentose phosphate pathway, which is used to produce nucleotides. Thus this event occurs at the expense of reduced glycolysis; how these cells compensate for this decrease in energy production is unknown. This gap in knowledge limits our ability to discover new therapeutic strategies to improve pregnancy outcomes. Our long-term goal is to understand the metabolic and molecular mechanisms responsible for endometrial stromal cell decidualization so that effective therapies for improving this process can be developed and used to prevent early pregnancy loss, especially in the increasing population of obese women. Here, we will test the central hypothesis that initiation of the cellular recycling pathway autophagy plays a key role in decidualization in both humans and mice. We will test our hypothesis by pursuing the following specific aims: Aim 1. Determine the role of autophagy in decidualization. Our working hypothesis is that decidualization depends on induction of autophagy both in vitro and in vivo. We will determine the effect of genetic or pharmacologic block of autophagy on decidualization of both mouse and human endometrial stromal cells. We will assess the effect of autophagy inhibition on both artificial and pregnancy-induced decidualization in a genetic knock out mouse model deficient in two different autophagic genes, Atg16L1 and Beclin1, allowing us to assay not only decidualization, but also embryo implantation and pregnancy outcomes. Aim 2. Define the function of GLUT8 in decidualization. We will test the hypothesis that the impaired decidualization and subfertility of Glut8 knockout mice are due to a loss of autophagy in endometrial stromal cells. We will address this by measuring autophagy in Glut8-null endometrial stromal cells and asking whether the defects can be reversed by inducing autophagy. We will also determine whether inducing autophagy can improve pregnancy rates in Glut8 knockout mice. Aim 3: Determine the mechanism by which saturated fatty acids impair decidualization. We will test our hypothesis that saturated free fatty acids inhibit autophagy by examining steps in autophagy and the signaling pathway that activates autophagy. We will also perform in vitro and in vivo mouse and human experiments to assess the ability of activators of autophagy to reverse the deleterious effects of high-fat diet, saturated fatty acids, and obesity on decidualization and reproductive outcomes. U[on completion of these aims we expected the following outcomes: 1) demonstration that autophagy is essential to optimal decidualization, and thus, implantation; 2) determination of the role of GLUT8 in decidualization; and 3) explanation of how diet can influence decidualization and implantation. This work will have an important impact on the fertility field by revealing mechanisms that are critical to successful decidualization, implantation, and overall pregnancy outcomes. This work will provide support for establishment of dietary guidelines and development of new therapeutics, such as autophagic activators, to possibly move into clinical trials to treat women with impaired implantation and pregnancy loss.

项目摘要 早期妊娠丢失影响了大约15%的已知妊娠，在肥胖者中可能更常见。 妇女早期妊娠丢失的一个突出原因是着床失败，这需要子宫 子宫内膜间质细胞经历蜕膜化。此外，最近的研究表明， 代谢调节在这个过程中。具体来说，蜕膜基质细胞产生较少的ATP， 更多的NADPH和核糖-5-磷酸通过戊糖磷酸途径，这是用来产生 个核苷酸因此，这一事件的发生是以减少糖酵解为代价的;这些细胞如何补偿这一点 能源生产的减少是未知的。这种知识上的差距限制了我们发现新的治疗方法的能力。 改善妊娠结局的战略。 我们的长期目标是了解子宫内膜异位症的代谢和分子机制， 基质细胞蜕膜化，从而可以开发用于改善该过程的有效疗法， 预防早孕流产，特别是在肥胖妇女人数不断增加的情况下。在这里，我们将测试 核心假设是细胞再循环途径自噬的启动在 人类和小鼠的蜕膜化。我们将通过追求以下具体目标来检验我们的假设： 目标1.确定自噬在蜕膜化中的作用。我们的假设是蜕膜化 依赖于体外和体内自噬的诱导。我们将确定遗传或 药物阻断小鼠和人子宫内膜间质细胞的自噬作用。我们 将评估自噬抑制对人工和妊娠诱导的蜕膜化的影响， 基因敲除小鼠模型缺乏两个不同的自噬基因，Atg 16 L1和Beclin 1，使我们能够 不仅测定蜕膜化，而且测定胚胎植入和妊娠结果。目标2.定义 GLUT 8在蜕膜化中的作用我们将检验以下假设： Glut 8基因敲除小鼠生育力低下是由于子宫内膜基质细胞中自噬的丧失。我们将解决 这是通过测量Glut 8缺失的子宫内膜间质细胞中的自噬，并询问这些缺陷是否可以 通过诱导自噬来逆转。我们还将确定诱导自噬是否可以改善妊娠 Glut 8基因敲除小鼠的死亡率。目的3：确定饱和脂肪酸损害的机制 蜕膜化我们将通过检测饱和游离脂肪酸抑制自噬来验证我们的假设。 自噬的步骤和激活自噬的信号通路。我们还将在体外和 体内小鼠和人实验以评估自噬激活剂逆转有害的自噬的能力。 高脂饮食、饱和脂肪酸和肥胖对蜕膜化和生殖结果的影响。U[on 完成这些目标，我们预期以下结果：1）证明自噬是必要的， 最佳的蜕膜化，因此，植入; 2）确定GLUT 8在蜕膜化中的作用;和3） 解释饮食如何影响蜕膜化和着床。这项工作将产生重要影响 通过揭示对成功的蜕膜化，植入和发育至关重要的机制， 整体妊娠结局。这项工作将为制定膳食指南提供支持， 开发新的治疗方法，如自噬激活剂，可能进入临床试验，以治疗 植入受损和妊娠丢失的妇女。