Developmental Requirements of SWI/SNF Chromatin Remodeling Complexes in Porcine Embryos

猪胚胎中 SWI/SNF 染色质重塑复合物的发育要求

• 批准号: 9322535
• 负责人: RYAN A CABOT
• 金额: $ 32.82万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9322535
• 关键词: Address; Affect; Age; Assisted Reproductive Techniques; Assisted Reproductive Technology; Biological Models; Cell Nucleus; Cells; Chromatin; Chromatin Remodeling Factor; Competence; Complex; DNA Methylation; Data; Development; Embryo; Embryonic Development; Enzymes; Epigenetic Process; Event; Failure; Family suidae; Fertilization in Vitro; Fetal Growth; Gene Expression; Genetic Transcription; Genome; Goals; Human; In Vitro; Individual; Infertility; Karyopherins; Laboratories; Lead; Link; Mediating; Methods; Oocytes; Pattern; Physiological; Population; Protein Subunits; Publishing; Regulator Genes; Reproductive Medicine; SMARCA4 gene; SWI/SNF Family Complex; Series; United States; Work; base; chromatin remodeling; design; early embryonic stage; embryo culture; embryo stage 2; experimental study; histone modification; improved; improved outcome; in vivo; overexpression; public health relevance; reproductive

 DESCRIPTION (provided by applicant): Infertility affects an estimated 6 million people in the United States, roughly 10% of the population of reproductive age. Treating human infertility can involve assisted reproductive technologies, which often include in vitro fertilization and embryo culture. It is well-established that mammalian embryos produced in vitro possess reduced developmental competence as compared to embryos produced in vivo. It is also known that embryos cultured in the laboratory can display epigenetic changes (both in DNA methylation and covalent histone modifications), aberrant gene expression and perturbed patterns of fetal growth. Despite the known links between in vitro embryo manipulation and perturbed epigenetic and transcriptional states, very little is known about the mechanisms by which epigenetic changes are mediated in cleavage stage embryos and how these epigenetic changes lead to developmental failure. To move toward the long term goal of improving the outcome for human embryos manipulated in vitro, two specific aims are addressed in this proposal; the aims are as follows: 1) Determine the mechanism by which SWI/SNF chromatin remodeling subunits access the nuclei of porcine oocytes and blastomeres of cleavage stage embryos, and 2) determine the developmental requirements of discrete SWI/SNF chromatin remodeling complex subunits in cleavage stage embryos. These aims will be addressed through a series of experiments that involve abalation and overexpression approaches to determine how regulatory components of the SWI/SNF chromatin remodeling enzymes are trafficked to the nuclei in cleavage stage embryos. The porcine embryo is used as the model systems because of its physiologic similarity to the human embryo during the early stages of embryonic development. The findings from these studies will have an impact on human reproductive medicine and will lead to development of improved methods for manipulating and culturing embryos.

 描述（由申请人提供）：在美国，不孕症影响着大约600万人，约占育龄人口的10%。治疗人类不育症可能涉及辅助生殖技术，其中通常包括体外受精和胚胎培养。众所周知，与体内产生的胚胎相比，体外产生的哺乳动物胚胎具有降低的发育能力。还已知在实验室中培养的胚胎可以显示表观遗传变化（DNA甲基化和共价组蛋白修饰）、异常基因表达和胎儿生长的扰动模式。尽管已知体外胚胎操作与扰动的表观遗传和转录状态之间存在联系，但对卵裂期胚胎介导表观遗传变化的机制以及这些表观遗传变化如何导致发育失败知之甚少。为了实现改善体外操作人类胚胎结果的长期目标，本提案提出了两个具体目标;目标如下：1）确定SWI/SNF染色质重塑亚基进入猪卵母细胞核和卵裂期胚胎卵裂球的机制，和2）确定卵裂期胚胎中分离的SWI/SNF染色质重塑复合物亚基的发育要求。这些目标将通过一系列涉及消融和过表达方法的实验来解决，以确定SWI/SNF染色质重塑酶的调节组分如何被运输到卵裂期胚胎的细胞核中。猪胚胎被用作模型系统，因为其在胚胎发育的早期阶段与人胚胎的生理相似性。这些研究的结果将对人类生殖医学产生影响，并将导致开发出更好的操作和培养胚胎的方法。