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

基本信息

批准号：
8097100
负责人：
CHARLES R LONG
金额：
$ 13.08万
依托单位：
TEXAS A&M AGRILIFE RESEARCH
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-07 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8097100
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

项目摘要

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，据估计，所有婴儿中约有1％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。已经实现了协助不育夫妇建立家庭的希望，但并非对这些革命技术对适当发展以及最终儿童健康和适应性的影响越来越关注。表观遗传编程失败是寻求无法解释的不育症，先天性异常以及增加与人类辅助生殖技术和自发概念相关的疾病的易感性的主要嫌疑人。表观遗传学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，而没有DNA序列的变化。两种主要的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被删除并随后在早期胚胎发育期间重新建立，这与生育诊所IVF后的胚胎培养期伴随着。我们研究的长期目标是在哺乳动物卵母细胞成熟，施肥和植入前的胚胎胚胎发育过程中开发出表观遗传调节的工作模型，以便可以修改体外胚胎处理系统以改善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在异常胚胎发育中表现出来。我们将通过使用RNA干扰来研究既定的体外胚胎发育模型中表观遗传重编程的功能基因组学来检验我们的假设。目的是评估沉默一组表观遗传调节剂对建立表观遗传标记的分子和生物学效应，维持多能量的标记和分化的启动。该目标将通过采用RNA干扰（RNAi）技术来静止调节植入前发育过程中调节表观遗传组的基因的表达，从而研究各个基因在表观遗传重编程和正常胚胎发育中的作用。将分析植入前发育过程中的全球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。该创新项目的成功完成将导致在早期胚胎的表观遗传重编程期间鉴定与维持和重建表观遗传程序有关的蛋白质。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：使用辅助生殖技术（例如体外受精）对不孕症的治疗一直很高，并且在美国迅速增长。许多研究表明，与这些程序相关的严重先天性异常和疾病的风险显着增加。与这些程序相关的表观遗传异常和疾病失败。表观遗传编程失败可能是指责的，因此了解表观遗传基因调节在胚胎发育过程中的作用对于制定降低不良结果可能性的不育治疗方案至关重要。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Oxygen-induced alterations in the expression of chromatin modifying enzymes and the transcriptional regulation of imprinted genes.

DOI：
10.1016/j.gep.2018.01.001
发表时间：
2018-06
期刊：
Gene expression patterns : GEP
影响因子：
0
作者：
Skiles WM;Kester A;Pryor JH;Westhusin ME;Golding MC;Long CR
通讯作者：
Long CR

Reshaping the transcriptional frontier: epigenetics and somatic cell nuclear transfer.

DOI：
10.1002/mrd.22271
发表时间：
2014-02
期刊：
MOLECULAR REPRODUCTION AND DEVELOPMENT
影响因子：
2.5
作者：
Long, Charles R.;Westhusin, Mark E.;Golding, Michael C.
通讯作者：
Golding, Michael C.

Examination of DNA methyltransferase expression in cloned embryos reveals an essential role for Dnmt1 in bovine development.

DOI：
10.1002/mrd.21306
发表时间：
2011-05
期刊：
MOLECULAR REPRODUCTION AND DEVELOPMENT
影响因子：
2.5
作者：
Golding, Michael C.;Williamson, Gayle L.;Stroud, Todd K.;Westhusin, Mark E.;Long, Charles R.
通讯作者：
Long, Charles R.

Down-regulation of viral replication by lentiviral-mediated expression of short-hairpin RNAs against vesicular stomatitis virus ribonuclear complex genes.

通过慢病毒介导的针对水泡性口炎病毒核糖核复合体基因的短发夹 RNA 表达下调病毒复制。