Characterizing effects of sperm- and oocyte-derived epigenetic factors on early embryonic gene expression and offspring metabolic function

精子和卵母细胞衍生的表观遗传因子对早期胚胎基因表达和后代代谢功能的影响

• 批准号: 9909811
• 负责人: Marina Krykbaeva
• 金额: $ 3.05万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-05 至 2023-01-04
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9909811
• 关键词: Address; Adult; Adult Children; Affect; Biological Models; Brown Fat; Cells; Communication; DNA; Development; Diet; Disease; Disease susceptibility; Educational process of instructing; Educational workshop; Embryo; Embryology; Embryonic Development; Ensure; Environment; Environmental Impact; Environmental Risk Factor; Epidemic; Epigenetic Process; Fathers; Fatty acid glycerol esters; Fellowship; Female; Fertilization in Vitro; Fetal Development; Fostering; Gene Expression; Genetic; Genetic Transcription; Genetic study; Germ Cells; Health; High Fat Diet; In Vitro; Individual; Inherited; Insulin Resistance; Knowledge; Life; Life Style; Link; Liver; Mammals; Massachusetts; Maternal-Fetal Exchange; Mentors; Metabolic; Metabolic Diseases; Metabolism; MicroRNAs; Microinjections; Modeling; Molecular; Morula; Mothers; Mus; Nutrient; Obesity; Oocytes; Pancreas; Parents; Parthenogenesis; Phenotype; Placenta; Population; Predisposition; Pregnancy; Prevention strategy; Protein-Restricted Diet; Proteins; RNA; Research; Risk; Science; Side; Skeletal Muscle; Small RNA; Stains; Stimulus; Surrogate Mothers; Techniques; Testing; Tissues; Training; Transfer RNA; Universities; Work; base; blastocyst; career development; differential expression; environmental stressor; experience; experimental study; genetic information; glucose tolerance; graduate student; improved; insight; insulin tolerance; intergenerational; male; medical schools; metabolic phenotype; mother nutrition; non-genetic; offspring; response; sperm cell; sperm function; symposium; transcriptome; transcriptome sequencing; transcriptomics; transmission process; zygote

PROJECT SUMMARY/ABSTRACT Metabolic diseases such as obesity have become significant health risks affecting one-third of the population worldwide and can have devastating complications. It is therefore imperative to understand the causes of metabolic disease predisposition in order to develop preventative strategies. Extensive genetic studies have failed to explain this ongoing epidemic. However, parents pass on not only genetic information, but also epigenetic factors, which can be modified in response to environmental stimuli, and can then affect gene expression. If information about parental environment can be recorded in the germline, it has the potential to be transmitted to the zygote and impact offspring health. This concept remains controversial in mammals and represents a large knowledge gap in the field of embryology. Previous work in the lab has demonstrated that sperm from fathers fed a low protein diet carry tRNA fragments and microRNAs that can modify gene expression in the embryo. Therefore, in Aim 1, sperm-derived RNAs will be purified and microinjected into parthenogenetically-activated oocytes, or parthenotes, which lack any paternal genetic content. Transcriptomic profiles of injected parthenotes will be acquired by single-embryo RNA-Seq to characterize resulting alterations to the embryonic transcriptome. Maternal transmission of epigenetic information has not been characterized to the same extent as the paternal side. Therefore, in Aim 2 a similar dietary paradigm will be utilized to address the question of whether information about maternal diet can be carried in oocytes and result in changes to the embryonic transcriptome. In vitro-fertilized embryos from mothers fed low protein, high fat, or control diet will be sequenced by single-embryo RNA-Seq. These embryos will be transferred to foster mothers to produce adult offspring, which will then be assessed for glucose tolerance and insulin resistance. The use of in vitro fertilization in this paradigm will ensure that any changes observed in the embryo originate from the oocyte and not from nutrient exchange during gestation. Completion of this research will elucidate effects of paternal and maternal epigenetic factors carried by gametes on the embryonic transcriptome. This work will be completed at the University of Massachusetts Medical School under the sponsorship of Dr. Oliver Rando. The fellowship training plan includes training in embryological techniques, such as microinjection and immunofluorescent staining of lineage markers, as well as metabolic phenotyping of adult mice. Opportunities to gain experience in science communication include participation in departmental seminars and local and national conferences. Furthermore, career development workshops are provided by the university, in addition to teaching and mentoring of first-year graduate students.

项目概要/摘要 肥胖等代谢性疾病已成为影响三分之一人口的重大健康风险 在世界范围内，可能会产生毁灭性的并发症。因此有必要了解其产生的原因 代谢疾病易感性，以便制定预防策略。广泛的遗传学研究表明 未能解释这种持续的流行病。然而，父母传递的不仅仅是遗传信息，还有 表观遗传因素，可以根据环境刺激进行修改，然后影响基因 表达。如果有关亲代环境的信息可以记录在种系中，那么它就有可能被记录在种系中。 传播到受精卵并影响后代的健康。这个概念在哺乳动物中仍然存在争议 代表了胚胎学领域的巨大知识差距。实验室之前的工作表明 低蛋白饮食的父亲的精子携带可以改变基因表达的 tRNA 片段和 microRNA 在胚胎中。因此，在目标 1 中，精子来源的 RNA 将被纯化并显微注射到 单性生殖激活的卵母细胞或单性生殖，缺乏任何父系遗传内容。 注射的单性动物的转录组图谱将通过单胚胎 RNA 测序获得 表征胚胎转录组的变化。表观遗传的母体传递 信息的特征还没有达到与父方相同的程度。因此，在目标 2 中，类似 将利用饮食范式来解决有关母亲饮食的信息是否可以 被携带在卵母细胞中并导致胚胎转录组的变化。体外受精胚胎 来自喂养低蛋白、高脂肪或对照饮食的母亲将通过单胚胎 RNA 测序进行测序。 这些胚胎将被转移给养母，以产生成年后代，然后将其 评估葡萄糖耐量和胰岛素抵抗。在这种范例中使用体外受精将 确保在胚胎中观察到的任何变化均源自卵母细胞，而不是源自胚胎发育过程中的营养交换 妊娠。这项研究的完成将阐明父本和母本表观遗传因素的影响 通过胚胎转录组上的配子。 这项工作将在马萨诸塞大学医学院在 Dr. 的赞助下完成。 奥利弗·兰多.奖学金培训计划包括胚胎学技术培训，例如显微注射 谱系标记的免疫荧光染色，以及成年小鼠的代谢表型分析。 获得科学传播经验的机会包括参加部门研讨会和 地方和国家会议。此外，大学还提供职业发展研讨会， 除了教学和指导一年级研究生外。