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）：子宫内的压力或出生后早期的压力在塑造未来健康方面起着基本作用。当胚胎对环境变化非常敏感，重新编程发育中的基因网络，组织和器官时，发生的初始压力源是出现的，从而使所产生的个体倾向于疾病。我们认为，次植入前的植入式条件使成年个体倾向于改变葡萄糖稳态，从而导致胰岛素抵抗的发生率增加。我们的初步数据表明，体外培养过程中应激增加会导致小鼠胚胎中基因表达的比例恶化。最重要的是，我们已经将硫氧还蛋白相互作用的蛋白（TXNIP）确定为植入式应激的标志。体外培养后胚泡中的TXNIP mRNA和蛋白质水平上调。比在优化培养基（氨基酸KAA，KAA）中培养的次优培养条件（Whitten's Medue-WM）的TXNIP上调更为强烈。成年动物和成年小鼠的TxNIP表达仍然升高，葡萄糖稳态受损。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，表明成年动物的植入前应激的记忆保持在植入前的胁迫。异常的TXNIP基因表达与控制DNA甲基化的几种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，一种钥匙细胞抗氧化剂。 TXNIP水平受葡萄糖，谷氨酰胺，含有化合物的腺苷的细胞含量调节。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AKT2/PKB2途径参与了外周葡萄糖代谢的调节，并且与胰腺β细胞量减少有关。我们建议（1）TXNIP是胚胎代谢感应机制的关键要素，并且（2）胚胎中的TxNIP上调修饰了稳态机制，因此成人组织中TXNIP将保持升高（3）成人组织中TXNIP升高导致glucose bolucose稳态的变化。为了检验这一假设，我们将研究：（1）TXNIP-TXN系统如何响应逐渐恶化的环境（2）负责维持成人组织中TxNIP表达升高的机制（3）成年小鼠体外产生的表型。这项研究很重要，因为将深入了解植入前胚胎利用的基本机制来应对不同的环境条件。此外，该研究得出的数据可用于监测通过辅助生殖技术（ART）构想的儿童。因此，此应用程序对计划公告（PA-08-104）响应：辅助生殖技术的不良结果。的确，在艺术的帮助下，已经构思了超过300万儿童，多项研究报告说，该人群的母体和胎儿并发症发生率增加。 公共卫生相关性：子宫内的压力或出生后早期的压力在塑造未来健康方面起着基本作用。当胚胎对环境变化非常敏感时，会发生一种初始的压力源，重新编程发育细胞，组织和器官，从而使所产生的个体易于疾病。这项研究旨在定义早期胚胎发育期间压力逐渐恶化的影响。从这项研究中获得的知识可能会导致人们对胚胎所使用的机制感知和适应不同环境的新知识。