Epigenetic Programming of Health and Disease in the Preimplantation Embyro

植入前胚胎健康和疾病的表观遗传编程

• 批准号: 9185992
• 负责人: PAOLO RINAUDO
• 金额: $ 32.89万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-12-01 至 2018-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9185992
• 关键词: Adult; Affect; Animals; Assisted Reproductive Technology; Atlases; Barker Hypothesis; Candidate Disease Gene; Catalogs; Cell Count; Cells; ChIP-seq; Child; Chromatin; Clinical; Collaborations; Conceptions; DNA Methylation; DNA analysis; Data; Data Set; Databases; Defect; Deposition; Development; Disease; Embryo; Embryonic Development; Environment; Environmental Risk Factor; Epigenetic Process; Evaluation; Fatty acid glycerol esters; Female; Fertilization; Fertilization in Vitro; Gene Expression; Generations; Genes; Genomics; Glucose Intolerance; Goals; Growth; Growth and Development function; Health; Heritability; Histones; Human; In Vitro; Infertility; Inner Cell Mass; Insulin; Laboratories; Lead; Light; Link; Liver; Metabolic; Metabolism; Methods; Mission; Mouse Strains; Mus; Outcome; Partner in relationship; Patients; Pattern; Phenotype; Physiological; Physiology; Placentation; Positioning Attribute; Process; Protocols documentation; Research; Resources; Sampling; Specimen; Stress; Testing; Time; Tissue Sample; Tissues; United States National Institutes of Health; Variant; Work; blastocyst; blood glucose regulation; career; cohort; embryo culture; epigenetic regulation; epigenome; epigenomics; genome-wide; genome-wide analysis; glucose tolerance; improved; in vivo; male; mature animal; metabolomics; mouse development; offspring; postnatal; preimplantation; programs; public health relevance; repository; response; sex; transcriptomics

 DESCRIPTION (provided by applicant): It is becoming increasingly clear that the preimplantation period is a time of genomic reprogramming. Preimplantation embryos may respond to an environmental factor by altering their developmental trajectory with the net results that their adult phenotype is altered. This observation has wide clinical implications, since in vitro fertilization (IVF) and preimplantation embryo culture are routinely used to treat patients with infertility and as of today, more than 5 million children have been conceived with assisted reproductive technologies (ART). The laboratory of the PI (Rinaudo) has devoted extensive resources to define the phenotype of mice following ART. We have found that 1) preimplantation embryos are sensitive to the culture conditions they are exposed to; 2) culture conditions more dissimilar to the physiologic in vivo conditions are associated with a more significant alteration of long term phenotypes; 3) glucose intolerance is a common phenotypic change present in adults generated by IVF; 4) using a candidate gene approach, selected chromatin alterations are present in preimplantation embryos and maintained in adult animals in a tissue specific and sexual dimorphic fashion. Although these data are valuable, multiple questions still remain unresolved. We still do not know what global epigenomic changes are present in embryos following ART and if these epigenetic changes persist in the adult. The laboratory of the co-PI (Ramalho-Santos) has an established track record on epigenetic regulation of early mouse development and the germline, with expertise in analyses of histone variants, histone marks and DNA methylation. They have recently developed methods for genome-wide analyses of histone marks using low numbers of cells. This proposal, in response to PAR 13-385, aims to leverage the expertise of both laboratories to determine how ART may program adult physiology. Our long term goal is to define how preimplantation culture conditions affect embryo development and postnatal growth, as a direct test to the Barker hypothesis. The specific goal of this application is to assess epigenetic changes in preimplantation embryos and adult tissues of control and IVF mice. The central hypothesis is that culturing preimplantation embryos in vitro will result in epigenetic changes in the blastocyst; these changes will persist in selected adult tissues in a sexual dimorphic fashion; the identified epigenetic changes might explain the observed adult phenotype. We plan to test our central hypothesis and thereby accomplish the objective of this application by analyzing DNA methylation and describing selected histone marks in the inner cell mass (ICM) of preimplantation embryos of different mouse strains generated in vivo or in vitro. Further, we will confirm if selected epigenetic marks persist in adult tissues. With respect to the expected outcomes, the work proposed is expected to provide a detailed reference epigenome of 1) preimplantation embryos and 2) adult tissue of mice conceived in vitro and in vivo.

 描述（由申请人提供）：越来越清楚的是，植入前时期是基因组重编程的时期，植入前胚胎可能通过改变其发育轨迹来响应环境因素，最终结果是其成年表型具有广泛的临床意义。由于体外受精 (IVF) 和植入前胚胎培养被常规用于治疗不孕症患者，截至目前，已有超过 500 万儿童通过辅助生殖技术受孕(ART). PI (Rinaudo) 实验室投入了大量资源来定义 ART 后的小鼠表型，我们发现 1) 植入前胚胎对它们所处的培养条件很敏感 2) 培养条件更加不同；体内生理条件与长期表型的更显着改变相关；3) 葡萄糖耐受不良是体外受精产生的成人中常见的表型变化；4) 使用候选基因方法，选择染色质改变存在于植入前胚胎中，并以组织特异性和性二态性方式维持在成年动物中，尽管这些数据很有价值，但仍有多个问题尚未解决，以及 ART 后胚胎中是否存在哪些整体表观基因组变化。这些表观遗传变化在成年体内持续存在，共同 PI (Ramalho-Santos) 的实验室在小鼠早期发育和种系的表观遗传调控方面拥有良好的记录，并具有组蛋白分析方面的专业知识。他们最近开发了使用少量细胞进行组蛋白标记的全基因组分析的方法，该提案是为了响应 PAR 13-385，旨在利用两个实验室的专业知识来确定 ART 的效果。我们的长期目标是确定植入前培养条件如何影响胚胎发育和出生后生长，作为对巴克假说的直接测试，该应用的具体目标是评估植入前的表观遗传变化。对照小鼠和 IVF 小鼠的胚胎和成年组织的中心假设是，体外培养植入前胚胎将导致囊胚发生表观遗传变化； 以性别二态性方式选择的成体组织；所识别的表观遗传变化可能解释观察到的成体表型，从而通过分析 DNA 甲基化和描述内细胞团中的选定组蛋白标记来实现本应用的目标。此外，我们将确认选定的表观遗传标记是否在成年组织中持续存在，所提出的工作预计将提供 1 的详细参考表观基因组。 ）植入前胚胎和 2) 体外和体内受孕的小鼠成年组织。

项目成果

From Embryos to Adults: A DOHaD Perspective on In Vitro Fertilization and Other Assisted Reproductive Technologies.

从胚胎到成人：体外受精和其他辅助生殖技术的 DOHaD 视角。

• 发表时间: 2016-08-09
• 作者: Feuer, Sky;Rinaudo, Paolo
• 通讯作者: Rinaudo, Paolo

Oxygen concentration alters mitochondrial structure and function in in vitro fertilized preimplantation mouse embryos.

氧气浓度改变体外受精植入前小鼠胚胎的线粒体结构和功能。

• 发表时间: 2019
• 作者: Belli, Manuel;Zhang, Ling;Liu, Xiaowei;Donjacour, Annemarie;Ruggeri, Elena;Palmerini, Maria Grazia;Nottola, Stefania Annarita;Macchiarelli, Guido;Rinaudo, Paolo
• 通讯作者: Rinaudo, Paolo