Effect of preimplantation embryo culture on adult glucose homeostasis

植入前胚胎培养对成人血糖稳态的影响

• 批准号: 8326737
• 负责人: PAOLO RINAUDO
• 金额: $ 30.78万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8326737
• 关键词: AKT2 gene; Acute; Adenosine; Adult; Adverse effects; Affect; Amino Acids; Animals; Antioxidants; Apoptosis; Assisted Reproductive Technology; Cell Proliferation; Cells; Child; DNA Methylation; Data; Defect; Deterioration; Disease; Down-Regulation; Elements; Embryo; Embryonic Development; Environment; Epigenetic Process; Future; Gene Expression; Genes; Glucose; Glutamine; Growth; Health; In Vitro; Incidence; Individual; Insulin Resistance; Knowledge; Lead; Link; Memory; Messenger RNA; Metabolic; Modeling; Monitor; Mus; NIH Program Announcements; Nutritional; Organ; Oxidation-Reduction; Oxidative Stress; PTEN gene; Pathway interactions; Peripheral; Phenotype; Play; Population; Proteins; Proto-Oncogene Proteins c-akt; Regulation; Reporting; Research; Role; Secondary to; Shapes; Stress; Structure of beta Cell of islet; System; Testing; Thioredoxin; Thioredoxin-2; Time; Tissues; Up-Regulation; adverse outcome; blastocyst; blood glucose regulation; design; embryo culture; environmental change; fetal; glucose metabolism; glucose transport; in utero; inhibitor/antagonist; insight; mature animal; novel; preimplantation; protein expression; public health relevance; stressor

DESCRIPTION (provided by applicant): Stress in utero or during the early post natal period plays a cardinal role in shaping future health. An initial stressor, occurring when the embryo is exquisitely sensitive to environmental changes, reprograms the developing gene networks, tissue, and organs so that the resulting individual is predisposed to disease. We hold that suboptimal preimplantation conditions predispose adult individuals to altered glucose homeostasis, leading to an increased incidence of insulin resistance. Our preliminary data show that increased stress during in vitro culture results in proportionate deterioration of gene expression in mouse embryos. Most importantly, we have identified thioredoxin-interacting protein (txnip) as a marker of preimplantation stress. Txnip mRNA and protein levels are upregulated in blastocyst after in vitro culture; suboptimal culture conditions (Whitten's medium- WM) results in a more intense txnip upregulation than culture in optimized medium (KSOM with amino acid, KAA). Txnip expression remains elevated in adult animals and adult mice have impaired glucose homeostasis. Significantly, adults animals cultured in WM have a more severe phenotype than animals cultured in KAA indicating that a memory of the preimplantation stress is maintained in the adult animal. The aberrant txnip gene expression is associated with altered expression of several genes that control DNA methylation, suggesting an epigenetic regulation. Txnip is the principal inhibitor of thioredoxin (txn), a key cellular antioxidant. Txnip levels are regulated by cellular contents of glucose, glutamine, adenosine containing compounds; txnip appears therefore to link the cellular nutritional state with redox state and subsequent metabolic regulation. In addition, txnip is involved in the regulation of peripheral glucose metabolism via downregulation of AKT2/PKB2 pathway and is associated with decreased pancreatic beta cell mass. We propose that (1) txnip is a key element of the embryonic metabolic sensing mechanism and that (2) txnip upregulation in the embryo modifies homeostatic mechanisms so that txnip will remain elevated in adult tissues (3) elevation of txnip in adult tissues results in alteration of glucose homeostasis. To test this hypothesis, we will investigate: (1) how the txnip-txn system responds to progressively worsening environment (2) the mechanisms responsible to maintain elevated txnip expression in adult tissues (3) the phenotype of adult mice generated in vitro. This study is important because will offer insight into basic mechanisms utilized by the preimplantation embryo to respond to different environmental conditions. In addition, data derived from this study can be used to monitor children conceived by assisted reproductive technology (ART). As such, this application is responsive to the Program Announcement (PA-08-104): Adverse outcome of Assisted Reproductive Technologies. Indeed, more than 3 million children have been conceived with the help of ART and multiple studies have reported an increased incidence of maternal and fetal complications in this population. PUBLIC HEALTH RELEVANCE: Stress in utero or during the early post natal period plays a cardinal role in shaping future health. An initial stressor, occurring when the embryo is exquisitely sensitive to environmental changes, reprograms the developing cells, tissues and organs so that the resulting individual is predisposed to disease. This research is designed to define how progressively worsening stress during early embryo development affect future health. Knowledge gained from this research may lead to novel understanding of the mechanisms used by the embryo to sense and to adapt to different environments.

描述（由申请人提供）：子宫内或纳塔尔早期的压力在塑造未来健康方面起着至关重要的作用。当胚胎对环境变化非常敏感时，最初的应激源会对发育中的基因网络、组织和器官进行重新编程，从而使产生的个体易于患病。我们认为，次优的植入前条件使成年个体易于改变葡萄糖稳态，导致胰岛素抵抗的发生率增加。我们的初步数据表明，在体外培养过程中增加压力会导致小鼠胚胎中基因表达的成比例恶化。最重要的是，我们已经确定了硫氧还蛋白相互作用蛋白（txnip）作为着床前应激的标志物。在体外培养后，Txnip mRNA和蛋白质水平在胚泡中上调;次优培养条件（Whitten培养基- WM）导致比在优化培养基（具有氨基酸的KSOM，KAA）中培养更强烈的Txnip上调。Txnip表达在成年动物中保持升高，并且成年小鼠具有受损的葡萄糖稳态。值得注意的是，在WM中培养的成年动物比在KAA中培养的动物具有更严重的表型，表明在成年动物中维持了植入前应激的记忆。异常的txnip基因表达与控制DNA甲基化的几个基因的表达改变有关，表明表观遗传调节。TXNIP是硫氧还蛋白（TXN）的主要抑制剂，硫氧还蛋白是一种关键的细胞抗氧化剂。Txnip水平受含葡萄糖、谷氨酰胺、腺苷的化合物的细胞内容物调节;因此Txnip似乎将细胞营养状态与氧化还原状态和随后的代谢调节联系起来。此外，txnip通过下调AKT 2/PKB 2途径参与外周葡萄糖代谢的调节，并与胰腺β细胞质量减少相关。我们提出：（1）txnip是胚胎代谢传感机制的关键元件，（2）txnip在胚胎中的上调改变了体内平衡机制，使得txnip在成体组织中保持升高;（3）txnip在成体组织中的升高导致葡萄糖体内平衡的改变。为了验证这一假设，我们将研究：（1）txnip-txn系统如何响应逐渐恶化的环境（2）负责维持txnip在成年组织中表达升高的机制（3）体外产生的成年小鼠的表型。这项研究很重要，因为它将为植入前胚胎对不同环境条件的反应提供基本机制。此外，从这项研究中获得的数据可用于监测通过辅助生殖技术（ART）受孕的儿童。因此，本申请是对计划公告（PA-08-104）的响应：辅助生殖技术的不良后果。事实上，在抗逆转录病毒疗法的帮助下，已经有300多万儿童受孕，多项研究报告说，这一人群中孕产妇和胎儿并发症的发病率增加。 公共卫生相关性：子宫内或纳塔尔早期的压力在塑造未来健康方面起着至关重要的作用。当胚胎对环境变化非常敏感时，最初的应激源会对发育中的细胞、组织和器官进行重新编程，从而使所产生的个体易于患病。这项研究旨在确定早期胚胎发育过程中逐渐恶化的压力如何影响未来的健康。从这项研究中获得的知识可能会导致对胚胎感知和适应不同环境的机制的新理解。