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.

项目总结/摘要 人类具有高度发达的认知能力和运动技能，这些特征反映在 我们的中枢神经系统（CNS）的体积扩大和细胞多样性。我的总体目标是侧写和比较 人类、非人类灵长类动物和啮齿类动物中的祖细胞多样性，从而确定 增加人类CNS的细胞多样性和大小。作为博士后研究的一部分，我收集并分析了 人类和小鼠脊髓运动的高时间分辨率单细胞RNA-seq数据 神经元（MN）谱系，这导致了分子上不同的，人类特异性的（即，中找不到 小鼠）运动神经元祖细胞（hsPMN）类型。我发现hsPMNs经历延迟和延长的 神经发生，使总MN输出增加~2倍。这项研究旨在描述他们如何 通过将单细胞RNA-seq与 长期星形胶质细胞共培养或谱系标记的人细胞的异种移植。第二，建议 本研究将通过以下方法进一步研究hsPMN细胞的进化及其基因表达程序：a）确定 是否在旧世界的猴子中发现了一种orthopathic细胞类型（即，猕猴），和B）鉴定和功能性地 测试新进化的基因调控元件，产生hsPMN特异性基因表达模式。 最后，我将开发一个3D脊髓类器官体外分化系统，该系统具有空间模式化的腹侧 和背脊髓谱系，使用可应用于人的标准分化条件， 猕猴和小鼠细胞。最后一个目标将扩大比较研究的范围， 多个脊柱谱系，从而为未来的研究开辟了新的途径和假设， 一个更全面的脊髓发育体外模型系统。洞察力、协议和数据 这项研究产生的结果将使人类神经祖细胞的研究成为可能，并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在CNS的大小和细胞多样性中发挥作用。 发展此外，这些研究和数据将构成我作为一名研究人员的研究议程的基础。 独立调查员，同时允许我扩大我的研究训练，包括不同的物种， 模型系统和广泛的实验以及计算技术。