Embryonic Gene Expression During Diabetic Embryopathy

糖尿病胚胎病期间的胚胎基因表达

• 批准号: 7578310
• 负责人: MARY R LOEKEN
• 金额: $ 35.32万
• 依托单位: JOSLIN DIABETES CENTER
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7578310
• 关键词: 1,2-diacylglycerol; ADP Ribose Transferases; Abbreviations; Affect; Antimycin A; Binding; Biochemical; Biochemical Pathway; Cells; Complications of Diabetes Mellitus; Congenital Abnormality; Development; Developmental Process; Diabetes Mellitus; Diabetic mother; Diabetic mouse; Diglycerides; Embryo; Embryonic Development; Esters; Funding; Gene Expression; Genes; Glucose; Glyceraldehyde; Heart; Hexosamines; Human; Hyperglycemia; Hypoxia; Inner Cell Mass; Lead; Malondialdehyde; Modeling; Molecular; Mothers; Mus; Neural Tube Closure; Neural Tube Defects; Neural tube; Neuroepithelial Cells; Oxidative Stress; PAX3 gene; Pathway interactions; Pregnancy; Pregnancy in Diabetics; Process; Protein Kinase C; Reactive Oxygen Species; Reduced Glutathione; Regulatory Element; Research; Stem cells; Stress; Structure; Superoxide Dismutase; System; Testing; activating transcription factor; adenylate kinase; cationic antimicrobial protein CAP 37; cell type; diabetic; diabetic embryopathy; dihydrofluorescein; embryonic stem cell; glucose metabolism; human embryonic stem cell; maternal diabetes; mouse model; prevent; research study; response; transcription factor

Project Summary: Diabetic embryopathy is a diabetic complication in which the early embryo of a mother with diabetes develops congenital malformations. The proposed experiments employ a mouse model of diabetic pregnancy in which expression of genes which control essential developmental processes is disturbed. These experiments focus on Pax3, a gene whose expression is significantly reduced in embryos of diabetic mice, which encodes a transcription factor required for development of structures that are often malformed during diabetic embryopathy, especially the neural tube and the heart. In the previous funding period, we showed that maternal hyperglycemia increases glucose delivery to the embryo, and that oxidative and hypoxic stress, resulting from excess glucose metabolism, causes impaired Pax3 expression, thereby leading to neural tube defects (NTD). In addition to experiments using embryos, we have used murine embryonic stem (ES) cells, which can be grown in quantity and can be induced to a Pax3-expressing neuroepithelial cell type. The central hypothesis to be tested in this proposal is that biochemical pathways affected by increased glucose metabolism by the early embryo causes oxidative and hypoxic stress, which prevents the induction or binding of transcription factors that activate expression of essential developmental control genes. In this proposal we will: (1) Investigate the interaction of hypoxic stress, oxidative stress, and increased protein kinase C activity leading to impaired Pax3 expression; (2) Identify factors which bind to a Pax3 regulatory element that is needed for differentiation-induced expression and which is inhibited by oxidative stress; and (3) Test the hypothesis that the same biochemical disturbances that lead to impaired Pax3 expression in mouse ES cells also lead to impaired PAX3 expression in human ES cells. Relevance: Understanding how maternal diabetes disturbs early embryonic development on a biochemical and molecular level is essential in order to devise new strategies to prevent them. The proposed research will use a mouse model of diabetic pregnancy and mouse embryonic stem cells to elucidate the biochemical mechanisms by which important embryonic gene expression is disturbed, and human embryonic stem cells to determine the extent to which the mouse and mouse stem cells serve as a model for the effects of biochemical disturbances during human diabetic pregnancy.

项目概述：糖尿病胚胎病是一种糖尿病并发症，其中母亲的早期胚胎 糖尿病患者会出现先天畸形所提出的实验采用小鼠模型， 糖尿病妊娠，其中控制基本发育过程的基因表达是 打扰这些实验集中在Pax 3上，这是一种在胚胎中表达显著减少的基因 它编码一种转录因子，这种转录因子是糖尿病小鼠结构发育所必需的， 在糖尿病胚胎病期间畸形，特别是神经管和心脏。在此前的融资中， 在此期间，我们发现母体高血糖增加了葡萄糖向胚胎的输送， 而由过量葡萄糖代谢引起的缺氧应激导致Pax 3表达受损，从而 导致神经管缺陷（NTD）。除了使用胚胎的实验外，我们还使用了小鼠 胚胎干（ES）细胞，其可以大量生长并且可以被诱导为表达Pax 3的细胞。 神经上皮细胞类型。 在这项提议中要检验的中心假设是，受增加的 早期胚胎的葡萄糖代谢引起氧化和缺氧应激，这阻止了诱导 或激活必需发育控制基因表达的转录因子的结合。在这 我们的建议是：（1）研究缺氧应激、氧化应激和蛋白质增加之间的相互作用 激酶C活性导致Pax 3表达受损;（2）鉴定与Pax 3调节因子结合的因子 分化诱导表达所需的并且被氧化应激抑制的元素;和 (3)检验以下假设，即导致Pax 3表达受损的相同生物化学紊乱， 小鼠ES细胞也导致人ES细胞中PAX 3表达受损。 相关性：了解母体糖尿病如何在生化方面干扰早期胚胎发育 而分子水平的研究对于制定新的预防策略至关重要。拟议研究 将使用糖尿病妊娠小鼠模型和小鼠胚胎干细胞来阐明 重要的胚胎基因表达受到干扰的机制，以及人类胚胎干细胞 以确定小鼠和小鼠干细胞作为模型的程度， 糖尿病妊娠期间的生化紊乱。