Elucidating molecular mechanisms regulatingspatiotemporal heterogeneity of microglia in the developing human brain

阐明调节人脑发育中小胶质细胞时空异质性的分子机制

• 批准号: 9816570
• 负责人: Galina Popova
• 金额: $ 6.16万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-13 至 2021-09-12
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9816570
• 关键词: Address; Affect; Age; Alzheimer' s Disease; Anatomy; Antibodies; BRAIN initiative; Bioinformatics; Biological; Brain; Brain region; Cell Communication; Cell Culture Techniques; Cells; Collaborations; Complex; Data; Development; Disease; Encephalitis; Environment; Event; Exhibits; Funding; Future; Gene Expression; Gene Expression Profile; Genetic Predisposition to Disease; Genetic Transcription; Heterogeneity; Heterotopic Transplantation; Homeostasis; Human; Individual; Laboratories; Light; Microglia; Microscopy; Modeling; Molecular; Molecular Analysis; Molecular Profiling; Morphology; Mus; Neuraxis; Neurodevelopmental Disorder; Neurosciences; Organogenesis; Pattern; Pharmacology; Population; Population Heterogeneity; Process; Role; Schizophrenia; Shapes; Slice; Symptoms; Testing; Therapeutic; Time; TimeLine; Tissue Sample; Tissues; Translating; Variant; Virus Diseases; Work; age related; autism spectrum disorder; base; cell type; induced pluripotent stem cell; innovation; insight; live cell imaging; nervous system disorder; neurogenesis; novel marker; programs; response; single-cell RNA sequencing; spatiotemporal; tissue culture; tool; transcriptomics

PROJECT SUMMARY/ABSTRACT In the developing human brain, microglia are a molecularly and morphologically heterogeneous population, interacting with different cell types and undergoing complex maturation programs in a region-specific manner. However, little is known about microglial maturation trajectories and how they are shaped by niche-specific intercellular interactions, hindering our understanding of their role in homeostasis and disease. Based on our preliminary finding that microglia in the developing brain exhibit transcriptional heterogeneity that defines microglia subpopulations, we propose that individual microglia subtypes elicit distinct functional and molecular responses on other cell types in the brain. Using single-cell RNA sequencing (scRNA-seq) in combination with live cell imaging and light-sheet microscopy, we will determine region-specific and age-dependent transcriptional signatures of developing microglia and address the long- standing question of whether these differences are determined by cell-intrinsic programs, or dictated by age- and microenvironment. We will address these questions in the following aims: Aim 1: Identify and validate region-specific molecular trajectories of microglia maturation in the developing human brain. We will identify regionally distinct maturation markers by examining gene expression differences in microglia from major anatomical regions, and then validate these markers using immunohistochemical and morphological analysis in primary tissue samples with light-sheet microscopy. Aim 2: Determine whether microglial regional identity affects brain development in a niche-dependent manner. We will perform heterotopic transplantations followed by unbiased scRNA-seq analysis to study the molecular consequences of microglia interactions with other cell types in a niche-specific manner. This work will achieve the following: (1) provide comprehensive transcriptional profiles of microglia across developmental stages and major anatomical regions in the developing human brain; (2) identify novel markers of microglial maturation and regionalization; (3) determine whether microglia are affected by the cell type composition of their microenvironment in a region-specific manner. Together, these findings will lead to a better understanding of microglial heterogeneity, help to refine existing iPSC-derived microglia models, and provide insight into how perturbations of microglia during development might contribute to neurological disorders such as autism and schizophrenia years later.

项目摘要/摘要 在发展中的人脑中，小胶质细胞是一种分子和形态上异质的人群， 与不同的细胞类型相互作用，并以特定区域的方式进行复杂的成熟程序。然而， 关于小胶质成熟轨迹及其如何通过小众特异性界面形状的知识知之甚少 相互作用，阻碍了我们对它们在体内平衡和疾病中的作用的理解。根据我们的初步发现 发育中的大脑中的小胶质细胞表现出定义小胶质细胞亚群的转录异质性，我们提出 该单个小胶质细胞亚型在大脑中的其他细胞类型上引起明显的功能和分子反应。使用 单细胞RNA测序（SCRNA-SEQ）与活细胞成像和灯表显微镜结合使用，我们将 确定开发小胶质细胞的区域特异性和依赖年龄的转录特征，并解决长期 这些差异是由细胞中的程序决定的，还是由年龄和年龄和 微环境。我们将在以下目的中解决这些问题： AIM 1：确定并验证发育中的人脑小胶质细胞成熟的区域特异性分子轨迹。 我们将通过检查主要的小胶质细胞中的基因表达差异来确定区域不同的成熟标记 解剖区域，然后在原代中使用免疫组织化学和形态分析验证这些标记 带有光片显微镜的组织样品。 目标2：确定小胶质区域身份是否以利基依赖性方式影响大脑发育。我们将 进行异位移植，然后进行无偏的SCRNA-SEQ分析，以研究 小胶质细胞与其他细胞类型的相互作用以特异性的方式与其他细胞类型相互作用。 这项工作将实现以下各种事情：（1）在整个环境中提供小胶质细胞的全面转录概况 发育阶段和发展中的人脑的主要解剖区域； （2）确定新颖的标记 小胶质成熟和区域化； （3）确定小胶质细胞是否受到细胞类型组成的影响 他们的微环境以特定区域的方式。这些发现一起将导致更好地理解 小胶质细胞异质性，有助于完善现有的IPSC衍生的小胶质细胞模型，并提供有关如何 发育过程中小胶质细胞的扰动可能会导致自闭症和诸如自闭症和 精神分裂症几年后。