Identifying human-specific neural progenitors and their role in neurodevelopment

识别人类特异性神经祖细胞及其在神经发育中的作用

• 批准号: 10662868
• 负责人: Sumin Jang
• 金额: $ 13.62万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10662868
• 关键词: 3-Dimensional; ATAC-seq; Achievement; Activities of Daily Living; Address; Adult; Amaze; Astrocytes; BRAIN initiative; Behavioral; Biological Assay; Biological Models; Catalogs; Cell model; Cell physiology; Cells; Central Nervous System; Cercopithecidae; Cerebral cortex; Characteristics; Coculture Techniques; Cognitive; Complex; Computational Biology; Computational Technique; Data; Development; Disease model; Dorsal; Evolution; Foundations; Gene Expression; Gene Expression Profile; Genetic Recombination; Goals; Human; In Vitro; Investigation; Label; Limb structure; Link; Macaca; Mammals; Molecular; Monkeys; Motor; Motor Neurons; Motor Skills; Mus; Nervous System; Neuroglia; Neuronal Differentiation; Neurons; Organoids; Output; Pathology; Pathway interactions; Pattern; Physiology; Pluripotent Stem Cells; Population; Population Heterogeneity; Population Sizes; Postdoctoral Fellow; Primates; Protocols documentation; Regulator Genes; Regulatory Element; Research; Research Personnel; Research Training; Rodent; Role; Running; Source; Specific qualifier value; Spinal; Spinal Cord; Standardization; System; Testing; Training; Transplantation; Vertebral column; Xenograft procedure; cell type; cognitive ability; comparative; human RNA sequencing; human model; in vitro Model; induced pluripotent stem cell; insight; nerve stem cell; neurodevelopment; neurogenesis; nonhuman primate; novel; progenitor; programs; single-cell RNA sequencing; stem cells; temporal measurement; tool; transcription factor

PROJECT SUMMARY/ABSTRACT Humans have highly advanced cognitive abilities and motor skills, characteristics which are reflected in the enlarged size and cell diversity of our central nervous system (CNS). My overall goal is to profile and compare progenitor cell diversity in humans, non-human primates and rodents, and thereby identify the origins of increased cell diversity and size of the human CNS. As part of my postdoctoral research, I collected and analyzed high-temporal-resolution single-cell RNA-seq data of the well characterized human and mouse spinal motor neuron (MN) lineage, which led to the identification of a molecularly distinct, human-specific (i.e., not found in mouse) motor neuron progenitor (hsPMN) cell type. I found that hsPMNs undergo delayed and protracted neurogenesis, increasing total MN output by ~2-fold. The proposed study aims to characterize how they contribute to the population size and subtype diversity of the MN lineage by combining single-cell RNA-seq with long-term astrocyte co-culture or xeno-transplantation of lineage-labeled human cells. Second, the proposed study will further investigate the evolution of hsPMN cells and their gene expression program by a) determining whether an orthologous cell type is found in old-world monkeys (i.e., macaque), and b) identifying and functionally testing newly evolved gene regulatory elements that give rise to hsPMN-specific gene expression patterns. Finally, I will develop a 3D spinal organoid in vitro differentiation system that harbors spatially patterned ventral and dorsal spinal lineages, using a standard set of differentiation conditions that can be applied to human, macaque, and mouse cells. This last aim will broaden the scope of comparative investigations to encompass multiple spinal lineages, thus opening up new avenues and hypotheses for future research, as well as providing a more comprehensive in vitro model system of spinal cord development. The insights, protocols, and data generated from this study will enable and provide valuable comparative analyses into human neural progenitor diversity and how human-specific progenitors contribute to the size and cell diversity of the CNS during development. Furthermore, these studies and data will form the foundation of my research agenda as an independent investigator, while allow me to broaden my research training to include a diverse host of species, model systems and a wide array of experimental as well as computational techniques.

项目摘要/摘要 人类具有高度高级的认知能力和运动技能，这反映在 我们中枢神经系统（CNS）的大小和细胞多样性的扩大。我的总体目标是概述和比较 人类，非人类灵长类动物和啮齿动物的祖细胞多样性，从而确定 人CN的细胞多样性和大小增加。作为博士后研究的一部分，我收集并分析了 良好表征人和小鼠脊柱电动机的高分辨率单细胞RNA-seq数据 神经元（MN）谱系，导致鉴定分子独特的人类特异性（即未在 小鼠）运动神经元祖细胞（HSPMN）细胞类型。我发现HSPMN经历了延迟并持久 神经发生，将总MN输出增加约2倍。拟议的研究旨在表征他们如何 通过将单细胞RNA-Seq与 谱系标记的人类细胞的长期星形胶质细胞共培养或异种转移。第二，提议 研究将进一步研究HSPMN细胞及其基因表达程序的演变，a）确定 是否在旧世界猴子（即猕猴）中找到直系同源的细胞类型，b）识别和功能 测试新进化的基因调节元件，从而引起HSPMN特异性基因表达模式。 最后，我将开发一个体外分化系统的3D脊柱机，该系统具有空间图案的腹侧 和背脊柱谱系，使用可以应用于人的标准分化条件集 猕猴和小鼠细胞。最后的目标将扩大比较调查的范围 多个脊柱谱系，从而为未来的研究开辟了新的途径和假设，并提供 脊髓发育的更全面的体外模型系统。见解，协议和数据 这项研究产生的将使并为人类神经祖细胞提供有价值的比较分析 多样性以及人类特异性祖细胞如何促进CNS的大小和细胞多样性 发展。此外，这些研究和数据将构成我的研究议程的基础 独立研究者，虽然让我扩大研究培训，以包括各种各样的物种，但 模型系统以及广泛的实验和计算技术。