Deciphering the Roles of Endometrial Glycogen Reserves and the Impact of Obesity on Fertility

解读子宫内膜糖原储备的作用以及肥胖对生育力的影响

• 批准号: 10638028
• 负责人: Matthew J Dean
• 金额: $ 37.29万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-08 至 2028-02-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10638028
• 关键词: AMHR2 gene; Address; Affect; Area; C-Peptide; C57BL/6 Mouse; Circadian Rhythms; Data; Decidua; Decidual Cell Reactions; Defect; Diestrus; Diet; Embryo; Embryonic Development; Endometrial; Endometrial Stromal Cell; Endometrium; Ensure; Enzymes; Epithelium; Estradiol; Fat-Restricted Diet; Fatty acid glycerol esters; Fertility; Fertilization; GYS1 gene; Global Change; Glucose; Glucose Transporter; Glycogen; Glycogen Phosphorylase; Glycolysis; Growth Factor; High Fat Diet; Hormonal; Human; Hyperglycemia; Hyperinsulinism; Hypoxia; Immunohistochemistry; Impairment; In Vitro; Infertility; Insulin; Invaded; Knock-out; Lactoferrin; Length; Link; Litter Size; LoxP-flanked allele; Metabolism; Modeling; Morula; Mus; National Institute of Child Health and Human Development; Nutrient; Obesity; Pathogenesis; Placentation; Pre-implantation Embryo Development; Pregnancy; Pregnancy Outcome; Pregnancy loss; Process; Proestrus; Progesterone; Regulation; Reproduction; Research; Role; Serum; Site; Spain; Time; Tissues; Uterus; Woman; blastocyst; diet-induced obesity; early pregnancy; early pregnancy loss; endometrial stroma; fasting glucose; glucose metabolism; glucose uptake; glucose-6-phosphatase; glycogen metabolism; human data; implantation; macromolecule; maternal obesity; metabolomics; mouse model; natural Blastocyst Implantation; novel; nutrition; pregnancy failure; preimplantation; reproductive; response; spatiotemporal; treatment group

Project Summary Over 40% of pregnancies fail in women, most before or during embryo implantation. During this time, embryos are dependent on secretions into the uterine lumen that contains all of the growth factors and nutrients necessary for embryo development. In particular, embryos need glucose. From fertilization until the morula stage, glucose uptake is low, but without glucose, the embryo degenerates. As embryos approach the blastocyst stage, glucose uptake increases dramatically, providing ATP to the embryo within the hypoxic uterine lumen. The endometrial stroma also uses a large amount of glucose to decidualize. After decidualization, the stroma relies on Warburg metabolism to produce ATP and supplies glucose to the invading embryo. Thus, it is clear that glucose availability must be regulated in a spatiotemporal manner to ensure a successful pregnancy. Finally, obesity leads to reduced fertility in women through, in part, effects on the uterus and its ability to support pregnancy. Most research on endometrial glucose has focused on the role of glucose transporters. However, the endometrium can also transiently store glucose as the macromolecule glycogen. In women, endometrial glycogen concentrations are correlated with fertility, but a direct link between uterine glycogen and fertility has never been established. To investigate this in mice, we collected uteri on proestrus and days post coitum (DPC) 1.5, 3.5, and 5.5. Our preliminary data show that the mouse endometrium stores two distinct glycogen pools: in the uterine epithelium and the uterine decidua. Epithelial glycogen peaked at proestrus and significantly reduced during the preimplantation period. The epithelial expressed glycogen phosphorylase and glucose-6-phosphatase, suggesting the epithelium can catabolize glycogen and secrete the resulting glucose. In contrast, the endometrial stroma stored little glycogen until decidualization, when glycogen content increased 7-fold. Similarly, artificially induced decidualization in hormonally primed mice resulted in a 5-fold increase in glycogen levels. In order to establish a definitive relationship between these distinct pools of glycogen and fertility, we obtained glycogen synthase 1 (GYS1) floxed mice so that we can knockout GYS1 in a tissue-specific manner. Aim 1 will generate an LTFiCre/+ GYS1flox/flox mouse to study the role of epithelial glycogen in glucose secretion and preimplantation embryo survival. Aim 2 will use AMHR2Cre/+ GYS1flox/flox mice to study the role of glycogen in the decidua. Collectively these aims will establish a causative link between the ability of the uterus to store glycogen in the epithelium and decidua and fertility. Aim 3 will use a diet-induced obesity model to study how maternal obesity affects glycogen metabolism in the mouse uterus. We will determine how a 45% and 60% fat diet affects uterine metabolism and fertility in a C57BL/6 mouse model. In vitro, we will determine the effects of hyperglycemia and hyperinsulinemia on decidual glycogen. In summary, this research will evaluate the role of endometrial glycogen stores in support of early pregnancy and assess the potential for maternal obesity to result in dysfunctional endometrial glycogen metabolism.

项目概要 超过 40% 的女性妊娠失败，大多数是在胚胎植入之前或期间。在此期间，胚胎 依赖于子宫腔内的分泌物，其中含有所有生长因子和营养物质 胚胎发育所必需的。特别是，胚胎需要葡萄糖。从受精到桑葚胚 阶段，葡萄糖摄取量低，但没有葡萄糖，胚胎就会退化。当胚胎接近 囊胚阶段，葡萄糖摄取急剧增加，为缺氧的胚胎提供ATP 子宫腔。子宫内膜间质也使用大量的葡萄糖来进行蜕膜。后 蜕膜化过程中，基质依靠 Warburg 代谢产生 ATP 并为入侵细胞提供葡萄糖 胚胎。因此，很明显，必须以时空方式调节葡萄糖的可用性，以确保 成功怀孕。最后，肥胖在一定程度上通过对子宫的影响导致女性生育能力下降 及其支持怀孕的能力。大多数关于子宫内膜葡萄糖的研究都集中在葡萄糖的作用上 运输者。然而，子宫内膜也可以暂时储存葡萄糖作为大分子糖原。在 对于女性来说，子宫内膜糖原浓度与生育能力相关，但子宫内膜糖原浓度与生育能力有直接联系。 糖原和生育力之间的关系尚未确定。为了在小鼠中研究这一点，我们收集了发情前期的子宫 和性交后天数 (DPC) 1.5、3.5 和 5.5。我们的初步数据表明，小鼠子宫内膜储存 两个不同的糖原库：子宫上皮和子宫蜕膜。上皮糖原达到峰值 发情前期和着床前期显着减少。上皮表达的糖原 磷酸化酶和葡萄糖-6-磷酸酶，表明上皮可以分解代谢糖原并分泌 产生的葡萄糖。相反，子宫内膜基质在蜕膜化之前储存很少的糖原，此时 糖原含量增加7倍。同样，在荷尔蒙引发的小鼠中人工诱导蜕膜化 导致糖原水平增加5倍。为了在这些之间建立明确的关系 由于糖原池和生育能力不同，我们获得了糖原合酶 1 (GYS1) floxed 小鼠，这样我们就可以 以组织特异性方式敲除 GYS1。目标 1 将生成 LTFiCre/+ GYS1flox/flox 小鼠来研究其作用 上皮糖原在葡萄糖分泌和植入前胚胎存活中的作用。目标 2 将使用 AMHR2Cre/+ GYS1flox/flox 小鼠研究糖原在蜕膜中的作用。总的来说，这些目标将建立一个因果关系 子宫在上皮和蜕膜中储存糖原的能力与生育能力之间的联系。目标 3 将使用 饮食诱导的肥胖模型，用于研究母亲肥胖如何影响小鼠子宫中的糖原代谢。 我们将确定 45% 和 60% 脂肪饮食如何影响 C57BL/6 小鼠的子宫代谢和生育能力 模型。在体外，我们将确定高血糖和高胰岛素血症对蜕膜糖原的影响。在 总之，本研究将评估子宫内膜糖原储备在支持早期妊娠和 评估母亲肥胖导致子宫内膜糖原代谢功能障碍的可能性。