Core C Transcriptomics

项目总结/摘要 Transcriptomic Core（C）提供关键的最先进的服务，以支持本提案中的所有项目。 核心C将应用全细胞和核单细胞转录组学来评估分子同一性，并激活 基因网络在特定的神经和非神经人脑细胞，并将使用尖端的“大面积” 空间转录组学（LaST），以绘制基因表达在细胞类型和大脑区域的关键阶段， 人类第三孕期新生儿脑发育。这些技术的使用将推动我们的 了解早产儿和足月新生儿大脑细胞谱系发育的机制， 作为一个高分辨率的分子图谱，以衡量如何围产期损伤，如缺氧 缺血性脑病（ischemic encephalopathy，HIE）影响脑的发育。此外，核心C将与项目 领导人开发了一系列新的细胞特异性标记物，用于已经定义的细胞类型，如神经细胞， 祖细胞，新生神经元，小胶质细胞和胶质细胞谱系，以及新的标志物， 作为识别和表征新生儿大脑的新描述的细胞类型的工具， 迁移的神经元流，在这个提议的最初任期内已经被发现。 核心C将由Kriegstein博士和Rowich博士共同指导，他们在人类大脑发育方面具有专业知识 以及分别在单细胞转录组学和空间转录组学中。克里格斯坦医生会监督 并指导日常操作、调度和实验设计。克里格斯坦博士的实验室 开发并验证了单细胞和单核mRNA测序技术，最近记录了 在与罗维奇实验室的两份联合出版物中（Velmeshev et al，Science，2019; Schirmer et al，Nature 2019年）; Rowich实验室为大面积空间转录组学（LaST）设计了一种独特的途径。 小鼠和人脑的绘图（Bayraktar等人，2019，Nat Neurosci; Schirmer等人，Nature 2019）;.的 这两种技术的结合使用将使项目1-3能够对不同的细胞类型进行分子分析 包括中间神经元，少突胶质细胞和小胶质细胞，发现新的细胞类型特异性标记，并绘制 通过单细胞mRNA测序鉴定的基因的原位表达模式。核心董事， Dmitry Velmeshev博士是一位才华横溢的生物信息学家，将担任核心经理， 为核心C制定了一个组织计划，旨在满足所有 项目他们还将协助PPG的项目负责人和学员，以优化设计， 执行他们的单细胞测序和绘图项目。此外，核心C将与科学 咨询委员会维持最高标准的质量控制，并与行政核心（核心 （一）管理业务和预算问题。核心C将提供关键支持，以帮助实现目标 项目1-3将共同提供对人类大脑发育的新见解， 新生儿发育阶段。

Project Summary/Abstract Transcriptomic Core (C) provides critical state-of-the-art services to support all of the Projects in this proposal. Core C will apply whole cell and nuclear single cell transcriptomics to evaluate molecular identity and activated gene networks in specific neural and non-neural human brain cells, and will use cutting-edge `Large area spatial transcriptomics (LaST)' to map gene expression across cell types and brain regions at key stages of human 3rd trimester-term neonatal brain development. The use of these technologies will advance our understanding of the mechanisms of cell lineage development in preterm and term neonatal human brain and serve as a high-resolution molecular map against which to measure how perinatal injuries, such as hypoxic ischemic encephalopathy (HIE) impact the development of the brain. Additionally, Core C will work with project leaders to develop a battery of novel cell-specific markers for already defined cell types, such as neural progenitor cells, newly born neurons, microglia, and glial cell lineages, as well as novel markers that will serve as tools to identify and characterize the newly-described cell types of the neonatal human brain such as the migrating neuronal streams that have already been discovered during the initial tenure of this proposal. Core C will be directed jointly by Drs. Kriegstein and Rowitch who have expertise in human brain development and in single cell transcriptomics and spatial transcriptomics respectively. Dr. Kriegstein will provide oversight and guidance of day-to-day operations, scheduling and experimental design. Dr. Kriegstein's laboratory has developed and validated both single cell and single nuclei mRNA sequencing technology recently documented in two joint publications with the Rowitch laboratory (Velmeshev et al, Science, 2019; Schirmer et al, Nature 2019); the Rowitch laboratory has designed a unique pathway for Large-area spatial transcriptomic (LaST) mapping of mouse and human brain (Bayraktar et al., 2019, Nat Neurosci; Schirmer et al, Nature 2019);. The combined use of both of these technologies will enable Projects 1-3 to molecularly profile diverse cell types including interneurons, oligodendrocytes, and microglia, discover new cell type specific markers, and map the in situ expression patterns of genes identified through single cell mRNA sequencing. The Core Directors, assisted by Dr. Dmitry Velmeshev, a talented bio-informatician who will serve as Core Manager, have developed an organizational scheme for Core C that is designed to meet the complementary needs of all the projects. They will also assist project leaders and trainees in the PPG in order to optimize design and execution of their single cell sequencing and mapping projects. In addition, Core C will work with the Scientific Advisory Committee to maintain the highest standard of quality control and with the Administrative Core (Core A) to manage operational and budgetary issues. Core C will provide critical support to help achieve the goals of Projects 1-3 that will, together, provide new insights into human brain development at clinically important neonatal stages of development.