BRD4-mediated regulation of human brain development and function

BRD4介导的人脑发育和功能调节

• 批准号: RGPIN-2022-03734
• 负责人: Tanaka, Yoshiaki
• 金额: $ 2.7万
• 依托单位: Université de Montréal
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=760604
• 关键词: BRD4; mediated; regulation; human; brain

My long-term research vision aims to study molecular regulations of brain development and functions. My research program applies two unique technologies: stem cell engineering and computational analysis that are promising approaches to generate various types of cells and tissues from individuals and handle large-scale biological data from the stem cell-derived cell types and tissues.  Brain is one of the most complex and important organs to control our emotional and behavioral processes. Brain is made up of multiple cell types, such as neurons, astrocytes, and microglia, that have their own unique properties and interact each other. Differentiation into each cell type is determined by the unique gene expression program. Histones are basic proteins that condense DNA into a compact form, called chromatin. Importantly, chemical modifications of histone proteins are known to contribute to the cell type-specific gene expression program by perturbing chromatin structure. Although more than 150 histone modifying enzymes have been reported so far, the most of their functional roles in brain development are largely unknown.  BRD4 is one of the modifying enzymes that are heavily expressed in brain and promote acetylation of lysine residues in histone proteins. We previously found that abnormal genomic binding of BRD4 impairs neuronal growth and electrical activation. It has been also reported that BRD4 also regulates glial cell differentiation and immune response of microglia. Although these data implicate that BRD4 is essential for development and function of both neuronal and glial cells in brain, it still remains unclear how BRD4 regulates cellular identity and distinct properties across multiple cell types.  We will differentiate human embryonic stem cells into neurons, astrocytes, and microglia with well-established protocols and investigate 1) genome-wide occupancy and 2) protein-protein interactions of BRD4 across different cell types using cutting-edge genomics and proteomics technologies. Furthermore, given the involvement of BRD4 in cell fate determination in brain, it is questionable 3) whether BRD4 also contributes structural organization of cell types in brain. To address this question, we will grow miniature brains from human embryonic stem cells with three-dimensional culture system and analyze whether BRD4 affects the organization of brain structure by high-resolution imaging analysis. Overall, the research program will employ in vitro tissue modeling and in silico analysis of large-scale datasets to shed light on fundamental epigenetic regulation of human brain development. Throughout the proposed project, students will obtain broad knowledge and expertise in stem cell biology, neuroscience, and bioinformatics.

我的长期研究愿景是研究大脑发育和功能的分子调控。我的研究项目应用了两项独特的技术：干细胞工程和计算分析，这是一种很有前途的方法，可以从个体中产生各种类型的细胞和组织，并处理来自干细胞来源的细胞类型和组织的大规模生物数据。大脑是控制我们情绪和行为过程的最复杂和最重要的器官之一。大脑由多种类型的细胞组成，如神经元、星形胶质细胞和小胶质细胞，这些细胞具有各自独特的特性，并相互作用。分化为每种细胞类型由独特的基因表达程序决定。组蛋白是一种基本蛋白质，可以将DNA浓缩成一种紧密的形式，称为染色质。重要的是，组蛋白的化学修饰通过干扰染色质结构而有助于细胞类型特异的基因表达程序。尽管到目前为止已经报道了150多种组蛋白修饰酶，但它们在大脑发育中的大多数功能在很大程度上是未知的。BRD4是一种修饰酶，在大脑中大量表达，促进组蛋白中赖氨酸残基的乙酰化。我们先前发现，BRD4的基因组结合异常会损害神经元的生长和电激活。也有报道BRD4还调节神经胶质细胞的分化和小胶质细胞的免疫反应。尽管这些数据表明BRD4对大脑中神经元和神经胶质细胞的发育和功能都是必不可少的，但仍然不清楚BRD4是如何调节多种细胞类型的细胞身份和不同性质的。我们将用成熟的方案将人类胚胎干细胞分化为神经元、星形胶质细胞和小胶质细胞，并利用尖端基因组学和蛋白质组学技术研究1)全基因组占有率和2)BRD4跨不同细胞类型的蛋白质-蛋白质相互作用。此外，鉴于BRD4参与了脑内细胞命运的决定，3)BRD4是否也参与了脑内细胞类型的结构组织是值得怀疑的。为了解决这个问题，我们将利用三维培养系统从人胚胎干细胞中培养出微型脑，并通过高分辨率成像分析来分析BRD4是否影响脑结构的组织。总体而言，该研究计划将采用体外组织建模和大规模数据集的电子分析，以阐明人类大脑发育的基本表观遗传调控。在整个拟议的项目中，学生将获得干细胞生物学、神经科学和生物信息学方面的广泛知识和专业知识。