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.

我的长期研究愿景旨在研究大脑发育和功能的分子调节。我的研究计划应用了两种独特的技术：干细胞工程和计算分析，这些技术是有望从个人生成各种类型的细胞和组织的方法，并处理来自干细胞衍生的细胞类型和组织的大规模生物学数据。大脑是控制我们的情感和行为过程的最复杂，最重要的器官之一。大脑由多种细胞类型组成，例如神经元，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相互作用。分化为每种细胞类型的分化是由唯一的基因表达程序确定的。组蛋白是基本蛋白质，将DNA凝结成一种称为染色质的紧凑形式。重要的是，已知组蛋白的化学修饰通过扰动染色质结构来促进细胞类型特异性基因表达程序。尽管到目前为止已经报道了150多个组蛋白修饰酶，但它们在大脑发育中的大部分功能作用在很大程度上尚不清楚。 BRD4是在大脑中大量表达并促进组蛋白中赖氨酸残留物的乙酰化的修饰酶之一。我们先前发现，BRD4的异常基因组结合会损害神经元的生长和电活激活。还报道了BRD4还调节了神经胶质，尽管这些数据表明BRD4对于脑中神经元和神经胶质细胞的开发和功能至关重要，但仍不清楚BRD4如何调节多种细胞类型的细胞身份和不同的特性。我们将通过良好的方案将人类胚胎干细胞分为神经元，星形胶质细胞和小胶质细胞，并研究1）全基因组占用率和2）使用最先进的基因组学和蛋白质组学技术在不同细胞类型上的蛋白质蛋白质相互作用。此外，鉴于BRD4参与大脑细胞脂肪的测定，这是值得怀疑的3）BRD4是否还促进了大脑中细胞类型的结构组织。为了解决这个问题，我们将使用三维培养系统从人类胚胎干细胞中增加微型大脑，并分析BRD4是否通过高分辨率成像分析影响大脑结构的组织。总体而言，该研究计划将采用体外组织建模和大型数据集的硅分析，以阐明人脑发育的基本表观遗传调节。通过拟议的项目，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。