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; 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.

项目摘要/摘要 肥胖等代谢疾病已成为影响三分之一人口的重大健康风险 在全球范围内，可能会有毁灭性的并发症。因此，必须了解 代谢疾病易感性，以制定预防策略。广泛的遗传研究 未能解释这种持续的流行病。但是，父母不仅传递遗传信息，而且传递 表观遗传因素可以根据环境刺激进行修改，然后会影响基因 表达。如果可以将有关父母环境的信息记录在种系中，则有可能 传输到合子并影响后代健康。在哺乳动物和 代表胚胎学领域的巨大知识差距。实验室的先前工作已经证明 父亲的精子喂养低蛋白质饮食携带tRNA片段和microRNA，可以改变基因表达 在胚胎中。因此，在AIM 1中，精子衍生的RNA将被纯化并微注射到 孤态激活的卵母细胞或parthenotes缺乏父亲的遗传含量。 注射parthenotes的转录组曲线将由单个Embryo RNA-Seq获取 表征对胚胎转录组的结果改变。表观遗传的母体传播 信息的表征与父亲方的特征程度相同。因此，在目标2中 饮食范式将用于解决有关孕妇饮食信息是否可以的问题 在卵母细胞中携带，并导致胚胎转录组的变化。体外施肥的胚胎 从喂养低蛋白的母亲中，高脂或对照饮食将通过单晶RNA-seq进行测序。 这些胚胎将被转移给寄养母亲，以产生成人后代，然后将是 评估葡萄糖耐受性和胰岛素抵抗。在此范式中使用体外受精将 确保在胚胎中观察到的任何变化源自卵母细胞，而不是来自营养的交换 妊娠。这项研究的完成将阐明父亲和母体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的赞助下完成。 奥利弗·兰多（Oliver Rando）。奖学金培训计划包括胚胎学技术的培训，例如显微注射 谱系标记物的免疫荧光染色以及成年小鼠的代谢表型。 获得科学沟通经验的机会包括参加部门研讨会和 地方和民族会议。此外，大学还提供职业发展研讨会 除了对一年级研究生的教学和指导。