Project 4: The Placenta-specific Glucose Transporter Modulation: obesity, metabol

项目 4：胎盘特异性葡萄糖转运蛋白调节：肥胖、代谢

• 批准号: 9520234
• 负责人: Johann Urschitz
• 金额: $ 24.76万
• 依托单位: UNIVERSITY OF HAWAII AT MANOA
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至 2020-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9520234
• 关键词: 20 year old; Address; Adult; Affect; American; Amino Acids; Attenuated; Biogenesis; Birth; Blood Circulation; Body Composition; Body mass index; Cardiovascular Diseases; Carrier Proteins; Cells; Centers of Research Excellence; Choriocarcinoma; DNA Polymerase II; DNA Transposable Elements; Developed Countries; Development; Discipline of obstetrics; Disease; Early Intervention; Epidemic; Epigenetic Process; Fetal Development; Fetal Diseases; Fetal Growth; Fetal Macrosomia; Fetus; Gene Targeting; Gene Transfer; General Population; Genes; Genetic; Genome; Glucose; Glucose Transporter; Goals; Health; High Fat Diet; Human; Immune response; Immunohistochemistry; Infant; Institutes; Insulin; Intervention; Lead; Life; Link; Liver; Longevity; Mediating; Metabolic; Metabolic Diseases; Metabolic syndrome; Metabolism; Methodology; Methods; Microbubbles; Minority; Modification; Mus; Mutation; Native Hawaiian; Neonatal; Nutrient; Obesity; Obesity associated disease; Organ; Overweight; Pacific Island Americans; Personal Satisfaction; Phase; Placenta; Plasma; Plasmids; Population; Predisposition; Pregnancy; Pregnancy Complications; Protein Biosynthesis; Protein Family; Protein Isoforms; Public Health; RNA; Reporter Genes; Reporting; Research; Role; SLC2A1 gene; Site; Syndrome; System; Testing; Tissues; Toxic effect; Transfection; Transgenes; Ultrasonography; United States; Virus; Weight; Western Blotting; Woman; adult obesity; base; fetal; fetal programming; gene therapy; glucose transport; high risk; improved; in utero; in vivo; insight; knock-down; maternal obesity; member; minimally invasive; minority health; mouse model; neonatal outcome; novel strategies; obese mothers; offspring; pregnant; promoter; protein expression; trophoblast

PROJECT SUMMARY/ABSTRACT - Project 4 The increase in obesity, particularly in industrialized countries has taken on epidemic proportions. In the US two-thirds of adults (20 years of age and older) are overweight or obese, with a body mass index or BMI above 25kg/m2. This epidemic extends to the pregnant population where more than half of all American women enter pregnancy with an increased BMI and as a consequence obesity is one of the most common high-risk obstetric syndromes. Maternal overweight and obesity are associated with disturbances in fetal growth leading to poor neonatal outcomes and a predisposition for cardiovascular disease and metabolic disorders later in life. The placenta serves as an interface between the fetal and maternal circulation and one of its key functions is to supply nutrients to the fetus. As the quantity of nutrients available to the fetus is a significant determinant of fetal growth, the placenta has been implicated in fetal overgrowth. The mechanisms linking in utero nutrient excess, fetal overgrowth and disease development later in life are poorly understood and are thought to include alterations in placental nutrient transport, genetic and epigenetic changes. We propose to develop a gene therapy approach to address this problem using a unique piggyBac transposon- based transfection system that was developed at our institute in Phase I of this COBRE. These GENIE plasmids are able to efficiently introduce genes into the host genome. Moreover, this non-viral approach presents many advantages over virus-based systems, including low toxicity and low immune response. We have recently employed a minimally invasive in vivo method (Ultrasound Targeted Microbubble Destruction, UTMD) for the delivery of GENIE plasmids achieving long-term expression of a reporter gene in the liver of mice. UTMD can mediate site-specific delivery of bioactive molecules to ultrasound-accessible target organs such as the placenta. Specifically, we will first attempt to knockdown the Glut1 expression in mice by stably introducing shRNAmir constructs into the genome of the placenta via UTMD. We will then determine the effects of Glut1 knockdown on placental glucose transport capacity on fetal growth and the metabolic syndrome in the offspring. We will also determine the effects of Glut1 knockdown on the function of primary human trophoblast (PHT) cells. Since the placenta is a tissue with a finite life span, essential for fetal development but discarded after birth, it is well suited for improving fetal wellbeing through genetic modifications, without causing problems associated with gene transfer directly into the host genome. The hypothesis that we will test in this application is that a placenta-specific reduction in Glut1 expression and consequent decrease of glucose transport into the placenta will attenuate fetal overgrowth and subsequent metabolic syndrome in offspring from dams fed a high-fat diet. Furthermore, placental gene transfer may serve as an early intervention strategy to reduce fetal overgrowth.

项目摘要/摘要 - 项目 4 肥胖症的增加，特别是在工业化国家，已达到流行病的程度。在 美国三分之二的成年人（20 岁及以上）体重指数或 BMI 超重或肥胖 25公斤/平方米以上。这种流行病蔓延到了孕妇，其中一半以上的美国人 女性怀孕时体重指数会增加，因此肥胖是最常见的问题之一 高危产科综合征。母亲超重和肥胖与胎儿发育障碍有关 生长导致新生儿结局不佳以及心血管疾病和代谢疾病的易感性 以后生活中的疾病。胎盘是胎儿和母体循环之间的界面，也是胎儿和母体循环之间的一个界面。 其主要功能是为胎儿提供营养。因为胎儿可获得的营养物质的量是 胎盘是胎儿生长的重要决定因素，与胎儿过度生长有关。机制 人们对子宫内营养过剩、胎儿过度生长和晚年疾病发展之间的联系知之甚少 并且被认为包括胎盘营养运输的改变、遗传和表观遗传的变化。我们 建议开发一种基因治疗方法，使用独特的piggyBac转座子来解决这个问题- 基于转染系统，该系统是我们研究所在 COBRE 第一阶段开发的。这些精灵 质粒能够有效地将基因引入宿主基因组。此外，这种非病毒式的方法 与基于病毒的系统相比，它具有许多优点，包括低毒性和低免疫反应。我们 最近采用了微创体内方法（超声靶向微泡破坏， UTMD）用于递送 GENIE 质粒，实现报告基因在肝脏中的长期表达 老鼠。 UTMD 可以介导生物活性分子向超声可到达的目标器官进行位点特异性递送 比如胎盘。具体来说，我们将首先尝试通过稳定地敲低小鼠中的 Glut1 表达 通过 UTMD 将 shRNAmir 构建体引入胎盘基因组。然后我们将确定 Glut1敲低对胎盘葡萄糖转运能力对胎儿生长和代谢的影响 后代综合症。我们还将确定 Glut1 敲低对初级功能的影响 人类滋养层（PHT）细胞。由于胎盘是一种寿命有限的组织，对胎儿至关重要 发育但在出生后被丢弃，它非常适合通过遗传改善胎儿的健康 修饰，不会引起与基因直接转移到宿主基因组相关的问题。这 我们将在本申请中测试的假设是 Glut1 表达的胎盘特异性减少和 随之而来的葡萄糖转运到胎盘的减少将减弱胎儿的过度生长和随后的生长 高脂肪饮食的水坝后代会出现代谢综合征。此外，胎盘基因转移可能有助于 作为减少胎儿过度生长的早期干预策略。