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.

我的长期研究目标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种独特的技术：干细胞工程和计算分析，这是从个体中产生各种类型细胞和组织的有前途的方法，并处理来自干细胞衍生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。大脑由多种细胞类型组成，如神经元，星形胶质细胞和小胶质细胞，它们具有自己独特的特性并相互作用。分化成每种细胞类型由独特的基因表达程序决定。组蛋白是一种碱性蛋白质，它将DNA浓缩成一种紧凑的形式，称为染色质。重要的是，已知组蛋白的化学修饰通过扰乱染色质结构而有助于细胞类型特异性基因表达程序。尽管迄今为止已经报道了超过150种组蛋白修饰酶，但它们在脑发育中的功能作用在很大程度上是未知的。 BRD 4是在脑中大量表达并促进组蛋白中赖氨酸残基乙酰化的修饰酶之一。我们先前发现BRD 4的异常基因组结合损害神经元生长和电激活。据报道，BRD 4还调节胶质细胞分化和小胶质细胞的免疫应答。尽管这些数据表明BRD 4对大脑中神经元和神经胶质细胞的发育和功能至关重要，但仍不清楚BRD 4如何调节多种细胞类型的细胞特性和不同特性。 我们将利用完善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，并使用尖端的基因组学和蛋白质组学技术研究1）全基因组占用和2）BRD 4在不同细胞类型中的蛋白质-蛋白质相互作用。此外，鉴于BRD 4参与脑中细胞命运的决定，3）BRD 4是否也有助于脑中细胞类型的结构组织是有疑问的。为了解决这个问题，我们将利用三维培养系统从人类胚胎干细胞中培养微型大脑，并通过高分辨率成像分析来分析BRD 4是否影响大脑结构的组织。总的来说，该研究计划将采用体外组织建模和大规模数据集的计算机分析，以揭示人类大脑发育的基本表观遗传调控。在整个项目中，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。