Altered metabolism in embryo generated by in vitro fertilization

项目总结/摘要 我们建议进行研究，以评估植入前胚胎操作对代谢的直接影响， 胚胎和组织。因为胚胎操作通常用于辅助生殖 技术（ART）治疗不孕症患者，这些目标具有广泛的临床意义。其实更多 800多万名儿童通过这些技术受孕，250多万名儿童通过体外受精受孕。 (IVF)周期每年都有。因此，有很大的需要知道，如果植入前胚胎操作， 对ART后代的健康产生微妙但可能持久的影响。 这个应用程序的重点是PI的专业知识的核心，因为他把大部分职业生涯都投入到 了解离体胚胎操作如何与胚胎的变化（基因改变）相关 表达、发育、线粒体功能）和后代的长期适应不良变化（缺陷 在胎盘形成中，改变了出生后的生长，并最终改变了葡萄糖稳态）。 虽然这些结果很重要，但仍有多个关键问题没有得到解答。特别是，分子 负责改变成人生长和代谢的机制尚不清楚。例如，目前尚不清楚， 在体外产生的胚胎中，代谢发生了改变，如果这些代谢改变在成年人中得以维持， 并负责导致IVF产生的成人葡萄糖处理异常。重要的是，总统 生物伦理学理事会和国会敦促NICHD确定这些不良后果是否 特别是与抗逆转录病毒疗法有关的。 令人兴奋的新的初步数据表明，糖酵解和乳酸代谢的改变是存在的， 胚胎和可能在IVF产生的成人中。根据初步数据，本申请的目的是 目的：1）了解糖酵解途径是否存在于经历不同程度糖酵解的胚胎中。 操纵;发现导致2）代谢改变和3）如果特定表观遗传的机制 最后，4）研究这些改变是否在IVF受孕的成年小鼠中保持。我们的中央 一种假设是，离体胚胎环境，与体内条件严重偏离（不同 氧浓度、改变的能量源）引起活性氧物质的增加。修复性 机制将诱导代谢改变和表观遗传变化。这些分子变化将是 在成年后代中维持，导致葡萄糖和乳酸盐代谢改变。关于预期 结果，我们将：1）确定什么样的培养条件下产生更大或更小的糖酵解变化， 胚胎，2）发现负责这些变化的途径，3）描述特定的表观遗传变化 诱导这些变化，并最终4）发现哪些代谢途径在IVF后代中发生了改变。 这些数据集预计将产生重要影响，因为它们对以下领域具有转化价值： 发育重编程、糖尿病、肥胖症和不孕症患者。

Project Summary/Abstract We propose studies to evaluate the direct impact of preimplantation embryo manipulation on the metabolism of embryos and tissues. Because embryo manipulation is routinely used in the context of Assisted Reproductive Technologies (ART) to treat patients with infertility, these goals have wide clinical implications. In fact, more than 8 million children have been conceived by the technologies and more than 2.5 million in vitro fertilization (IVF) cycles occur every year. Hence, there is great need to know if preimplantation embryo manipulation induces subtle, but possibly long-lasting effects on the health of ART offspring. The focus of this application is at the core of the PI’s expertise, as he has devoted most of his career to understanding how ex-vivo embryo manipulation is associated with changes in the embryo (altered gene expression, development, mitochondria function) and long-term maladaptive changes in the offspring (defects in placentation, altered postnatal growth and ultimately altered glucose homeostasis). While these results are important, multiple key questions remain unanswered. In particular, the molecular mechanisms responsible to alter growth and metabolism in adults are unknown. For example, it is unclear if metabolism is altered in embryos generated in vitro and if these metabolic alterations are maintained in adults and responsible to cause abnormal glucose handling in adults generated by IVF. Importantly, the President's Council on Bioethics and Congress urged the NICHD to determine whether these adverse outcomes are specifically related to ART procedures. Exciting novel preliminary data suggest that alteration of glycolysis and lactate metabolism are present in embryos and possibly in adult generated by IVF. Based on preliminary data, objectives of this application are to: 1) understand if alteration of glycolytic pathways exist in embryo who have undergone different degrees of manipulation; discover the mechanisms leading to 2) metabolic alterations and 3) if specific epigenetic changes exist; finally, 4) study if these alterations are maintained in adult mice conceived by IVF. Our central hypothesis is that the ex vivo embryonic environment, deviating profoundly from the in vivo conditions (different oxygen concentrations, altered energetic sources) cause increase in reactive oxygen species. Reparative mechanisms will induce metabolic alterations and epigenetic changes. These molecular alterations will be maintained in adult offspring resulting in altered glucose and lactate metabolism. Regarding expected outcomes, we will: 1) determine what culture conditions produce greater or smaller change in glycolysis in embryos, 2) discover the pathways responsible for these changes, 3) describe the specific epigenetic changes induced by these alterations and finally 4) discover what metabolic pathways are altered in IVF offspring. These datasets are expected to have an important impact because of the translational value to the fields of developmental reprogramming, diabetes, obesity and for patients affected with infertility.