Long-term physiological and behavioral outcomes, epigenetic profiles and multigenerational phenotypes in a mouse ART model

摘要 越来越多的不孕夫妇转向辅助生殖技术（ART）来治疗 他们的不孕症。越来越令人担忧的是，ART受孕的儿童患以下疾病的风险增加 特异性印记缺失疾病、先天性畸形、生长受限、先兆子痫 以及产后心脏和代谢紊乱。鉴于开展研究的困难， 使用人类胚胎，一种小鼠模型系统，该系统预期了与人类胚胎相关的一些风险 ART，将用于评估ART对胎盘形态学、印记基因 调节，生长，代谢，心脏和行为表型的后代，和基因 表达和染色质结构全基因组。具体目标1将评估表型， 包括生长、行为代谢和心血管功能， 与自然形成的对照相比。我们还将研究印记基因调控 和基因表达，DNA甲基化和染色质结构，使用基因特异性和高 吞吐量分析。此外，这一目标的设计将使我们能够分离，表型和 将胎盘与后代进行匹配，以确定胎盘表型是否可以准确地 预测体外受精（IVF）后代的健康状况。因为我们之前 报道了体外受精受孕后代的多种组织中表观遗传错误的低频率，我们 假设生殖细胞也含有表观遗传突变。在第二阶段，我们将 通过确定在IVF后代中观察到的异常表型是否 如果是的话，评估这种传播的机制。 这些实验的结果将提供有关这种联系的大量信息 通过ART受孕的后代的表观遗传变化和健康之间的关系， 表型可以预测后代的健康。我们的发现也可能表明实验性的 对ART程序的修改可以改善后代的结果。 !

Abstract Infertile couples are increasingly turning to Assisted Reproductive Technologies (ART) to treat their infertility. Of growing concern is that ART-conceived children are at increased risk for specific loss-of-imprinting disorders, congenital malformations, growth restriction, preeclampsia as well as postnatal cardiac and metabolic disorders. Given the difficulty of conducting studies using human embryos, a mouse model system, which anticipated some risks associated with ART, will be used to assess the effects of ART on placental morphology, imprinted gene regulation, growth, metabolic, cardiac and behavioral phenotypes of the offspring, and gene expression and chromatin structure genome-wide. Specific Aim 1 will assess the phenotypes, including growth, behavior metabolism and cardiovascular function, of ART-offspring in comparison to naturally-conceived controls. We will also interrogate imprinted gene regulation and gene expression, DNA methylation and chromatin structure using gene-specific and high throughput analyses. Moreover, the design of this aim will enable us to isolate, phenotype and match placenta to offspring to determine whether the placental phenotype can accurately predict health of in vitro fertilization (IVF)-derived offspring. Because we have previously reported a low frequency of epigenetic errors in multiple tissues of IVF-conceived offspring, we hypothesize that the germline cells also harbor epigenetic mutations. In Specific Aim 2, we will test this hypothesis by determining whether aberrant phenotypes observed in IVF offspring are transmitted to subsequent generations and, if so, assess the mechanism of this transmission. The result from these experiments will provide a trove of information regarding the linkage between epigenetic changes and health of offspring conceived by ART and whether placental phenotyping can predict offspring health. Our findings may also suggest experimental modifications to ART procedures that can improve offspring outcomes. !