Analysis of epigenetic regulation in early mammalian embryos via RNA interference

通过RNA干扰分析早期哺乳动物胚胎的表观遗传调控

• 批准号: 7755395
• 负责人: CHARLES R LONG
• 金额: $ 21.76万
• 依托单位: TEXAS A&M AGRILIFE RESEARCH
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-01-07 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7755395
• 关键词: Adopted; Age; Animal Model; Assisted Reproductive Techniques; Assisted Reproductive Technology; Back; Biochemical; Biological; Biological Assay; Biological Phenomena; Birth; Cattle; Cell Differentiation process; Cells; Child; Chromatin; Clinic; Cloning; Conceptions; Congenital Abnormality; Couples; Cryopreservation; DNA Methylation; DNA Methyltransferase; DNA Modification Methylases; DNA Sequence; Data; Development; Disease; Embryo; Embryonic Development; Environment; Enzymes; Epigenetic Process; Ethics; Event; Failure; Family; Fertility; Fertilization; Fertilization in Vitro; Gene Expression; Gene Expression Regulation; Gene Silencing; Gene Targeting; Genes; Genetic Engineering; Genome; Goals; Health; Histones; Human; In Vitro; Individual; Infertility; Intracytoplasmic Sperm Injections; Laboratories; Lead; Link; Literature; Maintenance; Measures; Methylation; Methyltransferase Gene; Modeling; Molecular; Mus; Nature; Outcome; Pattern; Physicians; Predisposition; Procedures; Production; Property; Proteins; Publishing; RNA Interference; Regulation; Regulator Genes; Research; Risk; Role; Study models; Suggestion; System; Techniques; Technology; Testing; Time; United States; Woman; Work; adverse outcome; base; blastocyst; chromatin modification; critical period; demethylation; embryo culture; embryo stage 2; fitness; functional genomics; histone modification; improved; in vitro Model; innovation; knock-down; oocyte maturation; pluripotency; preimplantation; pressure; programs; public health relevance; research study; response; social

DESCRIPTION (provided by applicant): In the United States, it is estimated that about 1% of all babies will be born using assisted reproductive technologies and these numbers continue to grow at a steady rate. The promise of assisting infertile couples to have a family has been realized, but not without a growing concern over the effects of these revolutionary technologies on the proper development and ultimately the health and fitness of the child. Failed epigenetic programming is a primary suspect in the search for causes of unexplained infertility, congenital abnormalities and increased susceptibility to disease associated with both human assisted reproductive techniques and spontaneous conception. Epigenetic refers to differential patterns of gene expression based solely on the physical and biochemical properties of chromatin, without a change in DNA sequence. Two major mechanisms appear to be responsible for establishing and maintaining the epigenome, DNA methylation and histone modifications. Mammalian epigenetic marks are first erased and subsequently re-established during early embryonic development, concomitant with the period of embryo culture following IVF in fertility clinics. The long-term goal of our research is to develop a working model of epigenetic regulation during mammalian oocyte maturation, fertilization and pre-implantation embryonic development, so that in vitro embryo handling systems can be modified to improve the outcomes following ART. Our hypothesis states that subtle alterations of histone and DNA methyltransferases during the critical period of in vitro embryo culture manifest in aberrant embryo development. We will test our hypothesis by using RNA interference to study the functional genomics of epigenetic reprogramming in an established model of in vitro embryo development. The goal is to evaluate the molecular and biological effects of silencing a select group of epigenetic regulators on the establishment of epigenetic marks, maintenance of markers of pluripotency and initiation of differentiation. This Aim will investigate the role individual genes in epigenetic reprogramming and normal embryonic development by employing RNA interference (RNAi) techniques to silence the expression of genes regulating the epigenome during pre-implantation development. Global chromatin methylation patterns and quantitative gene expression during pre-implantation development will be assayed to observe the response to gene silencing. The successful completion of this innovative project will result in the identification of the proteins involved with maintaining and re-establishing the epigenetic program during the period of epigenetic reprogramming in the early embryo. These experiments will provide the first evidence of the functional genes controlling the embryonic epigenome and their effects on cell differentiation in a mammalian species other than the mouse. PUBLIC HEALTH RELEVANCE: Treatment of infertility using assisted reproductive technologies such as in vitro fertilization is at an all time high and growing rapidly in the United States. Numerous studies show a significantly increased risk of serious congenital abnormalities and disease associated with these procedures. Failed epigenetic abnormalities and disease associated with these procedures. Failed epigenetic programming is likely to blame and thus understanding the role of epigenetic gene regulation during embryonic development is critical for formulating infertility treatment options that reduce the possibility of adverse outcomes.

描述（由申请人提供）：在美国，估计约1%的婴儿将使用辅助生殖技术出生，并且这些数字继续以稳定的速度增长。帮助不育夫妇建立家庭的承诺已经实现，但人们越来越担心这些革命性技术对儿童的正常发育以及最终对儿童的健康和体能的影响。失败的表观遗传编程是寻找原因不明的不孕症，先天性异常和增加与人类辅助生殖技术和自然受孕相关的疾病的易感性的主要嫌疑人。表观遗传是指基因表达的差异模式，仅基于染色质的物理和生化特性，而不改变DNA序列。两种主要机制似乎负责建立和维持表观基因组，DNA甲基化和组蛋白修饰。哺乳动物的表观遗传标记首先在早期胚胎发育过程中被消除，随后在生育诊所IVF后的胚胎培养期间重新建立。我们的研究的长期目标是开发一个工作模型的哺乳动物卵母细胞成熟，受精和植入前胚胎发育过程中的表观遗传调控，使体外胚胎处理系统可以修改，以改善ART后的结果。我们的假设指出，在体外胚胎培养的关键时期，组蛋白和DNA甲基转移酶的微妙变化表现在异常胚胎发育。我们将测试我们的假设，通过使用RNA干扰研究在体外胚胎发育的既定模型中的表观遗传重编程的功能基因组学。我们的目标是评估沉默一组选择的表观遗传调节因子对表观遗传标记的建立、多能性标记的维持和分化起始的分子和生物学效应。本研究的目的是通过RNA干扰（RNAi）技术沉默胚胎植入前发育过程中表观基因组调控基因的表达，研究单个基因在表观遗传重编程和正常胚胎发育中的作用。将测定着床前发育期间的整体染色质甲基化模式和定量基因表达，以观察对基因沉默的反应。这一创新项目的成功完成将导致在早期胚胎表观遗传重编程期间参与维持和重建表观遗传程序的蛋白质的鉴定。这些实验将提供控制胚胎表观基因组的功能基因及其对小鼠以外的哺乳动物物种细胞分化的影响的第一个证据。公共卫生相关性：在美国，使用辅助生殖技术（如体外受精）治疗不孕症的比例一直很高，而且增长迅速。许多研究表明，与这些手术相关的严重先天性畸形和疾病的风险显着增加。失败的表观遗传异常和疾病与这些程序。失败的表观遗传编程可能是罪魁祸首，因此了解胚胎发育过程中表观遗传基因调控的作用对于制定减少不良结果可能性的不孕症治疗方案至关重要。