Defining Characteristics of Cortical Progenitor Cells over Time in Mouse and Human

定义小鼠和人类皮质祖细胞随时间变化的特征

• 批准号: 9156213
• 负责人: SALLY TEMPLE
• 金额: $ 59.5万
• 依托单位: REGENERATIVE RESEARCH FOUNDATION
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-12-15 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9156213
• 关键词: Address; Adult; Architecture; Candidate Disease Gene; Cell Cycle; Cell division; Cells; Cerebral cortex; Cerebrum; Characteristics; Code; Complex; Data; Development; Disease; Embryo; Environmental Risk Factor; Generations; Genes; Heterogeneity; Human; Image Analysis; In Vitro; Knowledge; Life; Methods; Mus; Neurodegenerative Disorders; Neuroglia; Neurons; Pregnancy; Process; Production; RNA-Binding Proteins; Radial; Role; Specific qualifier value; Stem cells; Structure; Structure of choroid plexus; Testing; Time; Translations; Untranslated RNA; Vascular Endothelial Cell; Work; blastomere structure; daughter cell; developmental disease; in vivo; nerve stem cell; novel; progenitor; receptor; regenerative therapy; release factor; screening; stem cell technology; stem-like cell; therapy development; time use; transcriptome sequencing

Cerebral cortical development is a highly orchestrated process, with production of neurons destined for the cortical layers produced in order, deep to superficial, followed by glial generation. The timing of this process is very different between species. For example, mouse corticogenesis occurs over approximately a week of gestation, while in humans the process takes several months, resulting in a much larger and more complex cortex. Lineage studies have functionally defined the major types of neural progenitor cells (NPCs) contributing to corticogenesis, including stem cell-like radial glial cells (RGCs) and intermediate progenitor cells (IPCs). However, much remains to be discovered regarding how RGCs and IPCs are specified over time. We have discovered that during asymmetric RGC-IPC cell divisions, the RNA binding protein Stau2 segregates a complex cargo of coding and non-coding RNA specifically into the IPC daughter. Analysis of this cargo at different embryonic stages by RNA-sequencing has revealed networks of genes that are candidates for controlling proliferation and temporal specification of the IPC fate. Here we propose to test these candidates in functional studies, using high-throughput automated time-lapse image analysis for in vitro studies, as well as a novel lentiviral in vivo screening method, to define their roles in specifying IPCs and timing corticogenesis. In contrast to the progress made in understanding the characteristics of mouse cortical progenitor cells, less is understood regarding human cortical progenitors. Fundamental knowledge about how human RGCs and IPCs produce diverse progeny over time, their division mode, cell cycle times and lineages, remains unknown. Here we will address these gaps in knowledge using long-term time-lapse lineage analysis in vitro. In addition, by identifying genes expressed in human cortical progenitor cells, including at the single cell level and via analysis of the Stau2 cargo, we will reveal human cortical progenitor subtypes and heterogeneity. Further, a comparison of human and mouse cortical progenitor cell data will help illuminate key differences to address a major mystery: the difference in timing of mouse and human cortical development. Our lab continues to explore the interaction of environmental factors on cortical progenitor cells, aided by the ability to rapidly quantify changes in proliferation, division mode and differentiation using time-lapse analysis. Soluble factors released by structures in the germinal niche such as vascular endothelial cells and the choroid plexus, act on cortical progenitors to regulate the numbers and types of progeny they produce. Our recent work has identified a panel of candidate niche molecules secreted by the choroid plexus that could interact with receptors expressed on neural progenitors, which we propose to examine in vitro and in vivo, in mouse and human. Defining niche factors and their specific actions paves the way to address diseases that involve degeneration of stem cell zones which are normally active throughout life. Furthermore, defining environmental factors that act on human NPCs is important for translation towards regenerative therapy development.

脑皮质发育是一个精心策划的过程，其神经元的产生注定为按顺序产生的皮质层，深到浅表，然后是神经胶质产生。这个过程的时机在物种之间非常不同。例如，小鼠皮质发生发生在大约一周的妊娠期内，而在人类中，过程需要几个月的时间，导致皮层更大，更复杂。谱系研究在功能上定义了有助于皮质生成的主要类型的神经祖细胞（NPC），包括干细胞样径向神经胶质细胞（RGC）和中间祖细胞（IPC）。但是，关于如何随着时间的推移指定了RGC和IPC，还有很多尚待发现。我们发现，在不对称的RGC-IPC细胞分裂过程中，RNA结合蛋白STAU2将复杂的编码和非编码RNA的货物分离到IPC子中。通过RNA测序对该货物的分析揭示了基因网络，这些网络是控制IPC命运的增殖和时间规范的候选者。在这里，我们建议在功能研究中测试这些候选者，使用高通量自动化的延时图像分析，用于体外研究以及一种新型的体内慢病毒筛查方法，以定义其在指定IPC和时间定时皮质生成中的作用。与理解小鼠皮质祖细胞的特征方面所取得的进展相反，关于人皮质祖细胞的理解更少。关于人类RGC和IPC如何随着时间的推移产生不同的后代，其分裂模式，细胞周期时间和谱系的基本知识仍然未知。在这里，我们将在体外使用长期延时谱系分析来解决这些知识中的这些差距。此外，通过鉴定在人皮质祖细胞中表达的基因，包括在单细胞水平和通过对STAU2货物的分析，我们将揭示人类皮质祖细胞亚型和异质性。此外，人类和小鼠皮质祖细胞数据的比较将有助于阐明关键差异以解决一个主要的谜团：小鼠和人皮质发育的时间差异。我们的实验室继续探索皮质祖细胞上环境因素的相互作用，并在使用延时分析的能力迅速量化增殖，分裂模式和分化的能力的帮助下。结构在生发小裂中释放的可溶性因子，例如血管内皮细胞和脉络丛，作用于皮质祖细胞，以调节其产生的后代的数量和类型。我们最近的工作已经确定了一个脉络丛分泌的一组候选小众分子，该分子可以与在神经祖细胞上表达的受体相互作用，我们建议在小鼠和人类中检查体外和体内的体外和体内。定义利基因素及其特定行为铺平了解决涉及通常活跃的干细胞区域变性的疾病的方法。此外，定义对人NPC的环境因素对于转化为再生治疗的发展很重要。