Epigenetic Control of Development: Establishment and function of placenta-specific DNA methylation patterns

发育的表观遗传控制：胎盘特异性 DNA 甲基化模式的建立和功能

• 批准号: RGPIN-2021-02417
• 负责人: Hemberger, Myriam
• 金额: $ 4.23万
• 依托单位: University of Calgary
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2021
• 资助国家: 加拿大
• 起止时间: 2021-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=741965
• 关键词: Epigenetic; Control; Development; Establishment; function

My central research program examines the molecular processes that govern development of the placenta. My particular research focus is on how genes are regulated through instructive layers of information that are collectively known as the epigenome. I am aiming to understand how epigenetic modifications drive and reinforce cell fate decisions by establishing cell type-specific transcriptional networks that are of critical importance for developmental progression, ultimately ensuring reproductive success. DNA methylation is an epigenetic modification that is indispensable for embryonic development. The genomic distribution of DNA methylation in a cell lineage- and cell type-specific manner is key to reinforcing cell fate decisions, thereby ensuring stable differentiation. The placental trophoblast lineage is characterized by a unique DNA methylation profile that is highly distinct from that of embryonic cells. The establishment of this specific pattern is of crucial importance for trophoblast differentiation and normal placentation. Thus, mouse mutants that are defective in various components of the DNA methylation machinery or that lack specific methylation marks die early during development with severe placental abnormalities. Despite the pivotal importance of DNA methylation for placental development, it remains unknown how this epigenetic mark is targeted to specific sites in the genome to establish the distinct trophoblast-specific epigenomic landscape. This fundamental mechanistic question is the focus of my NSERC program. Since the DNA methylation machinery does not exhibit sequence-specificity by itself, the central hypothesis of this work is that a particular collection of DNA binding proteins unique to the trophoblast lineage confers the targeting of the DNA methylation machinery to particular sequence elements. The factors that combine sequence-specific DNA binding affinity with the attraction of DNA methylation components are called Kruppel-associated box zinc finger proteins (KRAB-ZFPs). This program of work is aimed at identifying critical KRAB-ZFPs that collectively establish the trophoblast methylome. The specific objectives are to: Aim #1: Identify and characterize trophoblast-enriched KRAB-ZFPs Aim #2: Establish the genomic occupancy profile of trophoblast KRAB-ZFPs Aim #3: Determine the capacity of KRAB-ZFPs to establish trophoblast-specific DNA methylation patterns Aim #4: Evaluate the impact of KRAB-ZFP ablation on trophoblast development Overall, this study will set milestones for our mechanistic understanding of how placental DNA methylation patterns are established, which is of fundamental importance for development and reproduction. Our HQP will learn cutting-edge stem cell and CRISPR gene editing techniques, bioinformatics analysis methods and epigenomic profiling technology, which will position them as leaders in Canada's natural sciences sector.

我的主要研究项目是研究控制胎盘发育的分子过程。我的特别研究重点是基因是如何通过被统称为表观基因组的有指导意义的信息层来调节的。我的目标是通过建立对发育进程至关重要的细胞类型特异性转录网络来了解表观遗传修饰如何驱动和加强细胞命运决定，最终确保生殖成功。DNA甲基化是胚胎发育过程中不可缺少的表观遗传修饰。DNA甲基化的基因组分布以细胞谱系和细胞类型特异性的方式是加强细胞命运决定的关键，从而确保稳定的分化。胎盘滋养细胞谱系的特点是独特的DNA甲基化谱，与胚胎细胞高度不同。这种特殊模式的建立对于滋养细胞分化和正常胎盘形成至关重要。因此，在DNA甲基化机制的各种组成部分中存在缺陷或缺乏特定甲基化标记的小鼠突变体在发育过程中早期死亡，并伴有严重的胎盘异常。尽管DNA甲基化对胎盘发育至关重要，但尚不清楚这种表观遗传标记如何靶向基因组中的特定位点，以建立独特的滋养细胞特异性表观基因组景观。这个基本的机械问题是我的NSERC项目的重点。由于DNA甲基化机制本身并不表现出序列特异性，因此这项工作的中心假设是，滋养细胞谱系特有的一组特定的DNA结合蛋白赋予了DNA甲基化机制对特定序列元素的靶向性。结合序列特异性DNA结合亲和力和DNA甲基化组分吸引力的因子被称为Kruppel-associated box zinc finger protein （KRAB-ZFPs）。本项目旨在鉴定关键的krab - zfp，这些关键的krab - zfp共同建立滋养层甲基化组。具体目标是：目标1：鉴定和表征滋养层细胞富集的KRAB-ZFPs；目标2：建立滋养层细胞KRAB-ZFPs的基因组占用谱；目标3：确定KRAB-ZFPs建立滋养层细胞特异性DNA甲基化模式的能力；目标4：总的来说，这项研究将为我们对胎盘DNA甲基化模式如何建立的机制理解树立里程碑，这对发育和生殖具有重要意义。我们的HQP将学习尖端的干细胞和CRISPR基因编辑技术，生物信息学分析方法和表观基因组分析技术，这将使他们成为加拿大自然科学领域的领导者。