Core C Transcriptomics

项目摘要/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来评估分子同一性和激活 在特定的神经和非神经人类脑细胞中建立基因网络，并将使用尖端的大面积 空间转录学(LAST)--绘制关键阶段跨细胞类型和大脑区域的基因表达图谱 人妊娠晚期新生儿脑发育。这些技术的使用将推动我们的 对早产儿和足月儿脑细胞谱系发育机制的认识 作为一个高分辨率的分子图谱，用来测量围产期损伤，如缺氧 缺血性脑病(HIE)影响大脑发育。此外，Core C将与项目配合使用 领导者为已经定义的细胞类型开发一系列新的细胞特异性标记，如神经细胞 祖细胞、新生神经元、小胶质细胞和神经胶质细胞谱系，以及将服务于 作为识别和表征新描述的新生儿人脑细胞类型的工具，例如 在这项提议的最初任期内已经发现的迁移神经元流。 CORE C将由Kriegstein博士和Rowitch博士联合执导，他们在人脑发育方面拥有专业知识 分别在单细胞转录组和空间转录组中表达。Kriegstein博士将提供监督 并指导日常运营、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 在与Rowitch实验室的两份联合出版物中(VelMeshev等人，科学，2019年；Schirmer等人，自然 2019年)；Rowitch实验室为大面积空间转录设计了一条独特的路径(上) 老鼠和人脑的图谱(Bayraktar等人，2019年，Nat Neurosci；Schirmer等人，自然，2019年)；这个 这两种技术的结合使用将使项目1-3能够对不同类型的细胞进行分子图谱分析 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测序确定基因的原位表达模式。各位核心董事， 在德米特里·维尔梅舍夫博士的协助下，一位才华横溢的生物信息学家将担任核心经理，他已经 制定了核心C的组织方案，旨在满足所有 项目。他们还将协助PPG中的项目负责人和受训人员，以优化设计和 执行他们的单细胞测序和测绘项目。此外，Core C将与Science 咨询委员会维持最高标准的质量控制和行政核心(核心 A)管理业务和预算问题。核心C将提供关键支持，帮助实现这些目标 项目1-3将共同为临床上重要的人脑发育提供新的见解 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.