Epigenetic and developmental regulation of embryonic plasticity and gametogenesis

项目概要/摘要：早期胚胎细胞是多能的，具有瞬时的能力， 产生一个有机体的所有细胞。发育生物学的一个基本问题是 这种暂时性发育可塑性的表观遗传因素，以及了解如何 实现并维持对特定细胞谱系的定型。胚胎多能性是如何建立的， 以及发育是否协调了细胞可塑性的限制与细胞命运的获得， 不太了解。表观遗传现象是指基因表达和染色质组织的变化 通过细胞分裂遗传而不改变潜在的DNA序列。最近，在我的研究 博士后研究发现了一个不受重视的发展规律， 表观遗传信息的分子载体，复制偶联组蛋白H3和组蛋白变体H3.3， 在配子发生过程中，影响早期胚胎的表观遗传组织，对 多能性和谱系承诺。 这一独立之路奖提案的目标是请求对培训的支持， 发展专门知识，并应用，基因组方法，同时解决表观遗传调控的作用， 胚胎发育早期的可塑性，以及细胞命运的维持。在此期间支持K99/R 00 我的职业生涯的一个阶段将是我作为一个独立研究人员的成功发展的变革。的 本提案中概述的研究计划和职业发展将利用广泛的 约翰霍普金斯大学的资源，以及与基因组学和 生物信息学这项提案将研究表观遗传调控和发育时间的早期 胚胎可塑性具体而言，在K99指导阶段，该奖项的主要研究者（PI） 我将联合收割机结合目前的专业知识和在约翰霍普金斯大学博士后培训期间产生的初步结果 大学，与新开发的技术开发的合作者的实验室，以回答 关于胚胎可塑性和细胞增殖的动态表观遗传机制的基本问题 命运限制这种方法有望解决以下具体目标：1）确定发展的 H3和H3.3双峰掺入在早期胚胎发生和配子发生中的意义，2） 表征H3，H3.3的全基因组定位及其在早期免疫过程中的翻译后修饰， 胚胎发生，以及它们对染色质可及性和基因表达的影响，以及3）描绘 协调两类不同组蛋白基因发育调控的分子途径 对生育和细胞命运限制很重要。完成这项培训将发展我的研究专长， 同时描绘胚胎可塑性的表观遗传调控机制，这将是 适用于生殖生物学、胚胎学、细胞重编程和生育领域。

Project Summary/Abstract: Early embryonic cells are pluripotent, possessing the transient capacity to generate all the cells of an organism. A fundamental question in developmental biology concerns identifying the epigenetic factors that underlie this temporary developmental plasticity, as well as understanding how commitment to a specific cell lineage is achieved and maintained. How embryonic pluripotency is established, and whether development coordinates the restriction of cellular plasticity with the acquisition of cell fate is poorly understood. Epigenetic phenomena refer to changes in gene expression and chromatin organization inherited through cell divisions without changing the underlying DNA sequences. Recently, studies during my postdoctoral research have uncovered an unappreciated developmental regulation of the incorporation of key molecular carriers of epigenetic information, the replication-coupled histone H3 and histone variant, H3.3, during gametogenesis that influences the epigenetic organization in the early embryo, with a lasting effect on pluripotency and lineage commitment. The goal of this Pathway to Independence Award proposal is to request support for training in order to develop expertise in, and to apply, genomic approaches while addressing the role of epigenetic regulation of plasticity during early embryonic development, as well as cell fate maintenance. K99/R00 support during this stage of my career will be transformative to my successful development as an independent researcher. The research plans and career development outlined in this proposal will take advantage of the extensive resources at Johns Hopkins University, as well as collaborate with leading experts in genomics and bioinformatics. This proposal will examine the epigenetic regulation and developmental timing of early embryonic plasticity. Specifically, during the K99 mentored phase of this award the principal investigator (PI) will combine current expertise and preliminary results generated during postdoctoral training at Johns Hopkins University, with newly developed techniques developed in the collaborator's laboratories to answer fundamental questions regarding the dynamic epigenetic mechanisms underlying embryonic plasticity and cell fate restriction. This approach promises to resolve the following specific aims 1) determine the developmental significance of H3 and H3.3 bimodal incorporation during early embryogenesis and gametogenesis, 2) characterize the genome-wide localization of H3, H3.3, and their post-translational modifications during early embryogenesis, as well as their influence on chromatin accessibility and gene expression and 3) delineate the molecular pathways that orchestrate the developmental regulation of two distinct classes of histone genes important for fertility and cell fate restriction. The completion of this training will develop my research expertise, while delineating the mechanisms underlying the epigenetic regulation of embryonic plasticity, which will be applicable to the fields of reproductive biology, embryology, cellular reprogramming, and fertility.