MOLECULAR AND METABOLIC ASPECTS OF IMPLANTATION

植入的分子和代谢方面

基本信息

批准号：
8064306
负责人：
KELLE H MOLEY
金额：
$ 49.26万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-05-01 至 2015-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8064306
关键词：
Affect Androgens Animal Model Area Belief Biological Assay Biology Cell Culture System Cells Cellular biology Characteristics Clinical Data Decidual Cell Reactions Development Endocrine System Diseases Endometrial Endometrial Stromal Cell Endometrium Estradiol Estrogens Event Exposure to Failure Functional disorder Genetic Glucose Glucose Transporter Gonadal Steroid Hormones Hormones Human Hyperandrogenism In Vitro Insulin Resistance Intervention Laboratories Link Measures Mediating Metabolic Metabolic Pathway Molecular Mus Obesity Oocytes Pathway interactions Patients Pattern Pentosephosphate Pathway Physiological Polycystic Ovary Syndrome Population Pregnancy Process Progesterone Research SLC2A1 gene Sampling Site Small Interfering RNA Specimen Spontaneous abortion Subfamily lentivirinae Testing Therapeutic Intervention Time Uterus Woman dehydroepiandrosterone effective therapy endometrial stroma enzyme activity experience failure Implantation glucose metabolism glucose transport glucose uptake hormone regulation human embryonic stem cell implantation in vitro Model in vivo in vivo Model innovation mRNA Expression mouse model natural Blastocyst Implantation novel novel strategies novel therapeutic intervention patient population pregnant prevent protein expression public health relevance pup reproductive small hairpin RNA

项目摘要

DESCRIPTION (provided by applicant): This study will be one of the first attempts to link the increased levels of estradiol and androgens/decreased levels of progesterone with miscarriages seen in women with PCOS to a mechanism which focuses on aberrant glucose utilization. Understanding the hormonal regulation of glucose uptake and metabolism in the uterine stroma during decidualization will potentially progress into the development of novel pharmacologic interventions to increase the rate of successful pregnancies in this population of women. This proposal is organized into three specific aims. In Aim 1, we will investigate the effects of high levels of estradiol/androgens and low levels of progesterone on expression of glucose transporters, glucose utilization and decidualization in murine endometrial stromal cells. We will also perform the same studies in human endometrial stromal cells. We hypothesize that high E2 and low P4 will downregulate GLUT1 and glucose uptake in mESCs and hESCs. In Aim 2, we will analyze the effects of changing GLUT1 expression as well as altering glucose metabolism on the process of decidualization of these cells. We will use siRNA/shRNA lentivirus to knockdown GLUT1, glucose flux studies to assess glucose utilization in these cells and then inhibit these pathways and assess decidualization in ESCs. We hypothesize that glucose is primarily metabolized via the pentose phosphate pathway during decidualization; and that inhibition either upstream (GLUT1 expression) or downstream (PPP inhibition) adversely affects decidualization. We will test this hypothesis in mouse and human ESCs. Finally, in Aim 3 we will focus on analyzing the in vivo effects of excess estradiol/androgen on decidualization and glucose utilization in the endometrial stroma by using a mouse model of increased DHEA. We will attempt to correct these effects by exposure to PPP activators used both in vivo and in vitro. We will also use leftover samples from patients with PCOS or oocyte donor patients and examine human ESCs. We predict that 1) DHEA exposed mice will have abnormal glucose utilization and thus abnormal decidualization with decreased numbers of pups, 2) the agents used to increase glucose utilization via the PPP will reverse decidualization abnormalities, and 3) human ESCs from these populations of patients will have similar abnormalities in glucose utilization. PUBLIC HEALTH RELEVANCE: Women with polycystic ovary syndrome (PCOS), as well as other endocrine disorders with elevated estradiol or androgens, all experience embryo implantation failure and endometrial dysfunction. The general belief is that this is due to the effects of excess estrogen/androgen or progesterone deficiency, both characteristic of this patient population. Our hope is that as a result of our studies examining the effects of high estrogen/androgen levels on glucose utilization in the uterus, new therapeutic interventions for implantation failure in patients with PCOS, and these endocrine disorders, may be discovered.

描述（由申请人提供）：这项研究将是将雌二醇和雄激素/雄激素降低水平升高与PCOS女性中的流产联系起来的首次尝试之一，以将重点放在异常葡萄糖利用率上。了解在划分过程中子宫基质中葡萄糖摄取和代谢的激素调节可能会进展为新的药理学干预措施的发展，以提高这一女性人群的成功妊娠率。该提议分为三个具体目标。在AIM 1中，我们将研究高水平的雌二醇/雄激素以及低水平的孕激素对葡萄糖转运蛋白表达，葡萄糖利用率和在鼠子宫内膜基质细胞中的表达的影响。我们还将在人子宫内膜基质细胞中进行相同的研究。我们假设高E2和低P4将在MESC和HESC中下调GLUT1和葡萄糖摄取。在AIM 2中，我们将分析改变GLUT1表达的影响，并改变葡萄糖代谢对这些细胞的脱字过程的影响。我们将使用siRNA/SHRNA慢病毒来敲除GLUT1，葡萄糖通量研究，以评估这些细胞中的葡萄糖利用，然后抑制这些途径并评估ESC中的deDIALIDIAD。我们假设葡萄糖主要是通过脱牙期间通过磷酸五磷酸途径代谢的。这种抑制在上游（GLUT1表达）或下游（PPP抑制）会不利地影响义词化。我们将在小鼠和人类ESC中检验这一假设。最后，在AIM 3中，我们将专注于通过使用增加的DHEA小鼠模型来分析过量雌二醇/雄激素对子宫内膜基质中脱核和葡萄糖利用的体内影响。我们将尝试通过暴露于体内和体外使用PPP激活剂来纠正这些效果。我们还将使用PCOS或卵母细胞供体患者患者的剩余样品并检查人类ESC。我们预测1）DHEA暴露的小鼠将具有异常的葡萄糖利用率，因此幼体数量减少，幼虫数量减少，2）通过PPP增加葡萄糖利用的药物将反向脱义异常，3）患者中的人类ESC的这些人群会在这些范围内造成类似的异常症状化。公共卫生相关性：多囊卵巢综合征（PCOS）以及其他内分泌疾病患有雌二醇或雄激素升高的妇女，所有这些都经历了胚胎植入衰竭和子宫内膜功能障碍。人们普遍认为，这是由于该患者人群的特征过多的雌激素/雄激素或孕激素缺乏症的影响。我们的希望是，由于研究了高雌激素/雄激素水平对子宫中葡萄糖利用的影响，可能会发现PCOS患者植入衰竭的新治疗干预措施以及这些内分泌疾病。