Human Correlation Core

人类相关核心

• 批准号: 10240686
• 负责人: Richard J Ulevitch
• 金额: $ 34.93万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10240686
• 关键词: ATAC-seq; Blood Cells; CD34 gene; CRISPR/Cas technology; Cell Lineage; Cells; Chromatin; Cytometry; Data; Funding; Gene Expression Regulation; Genes; Genetic study; Human; Immune; Immunity; Infection; Inflammation; Laboratories; Measurement; Measures; Mediating; Messenger RNA; Methods; Molecular; Mus; Myeloid Cells; Network-based; Orthologous Gene; Pathway Analysis; Pathway interactions; Protocols documentation; Regulator Genes; Research Personnel; Role; Sampling; Seminal; Signal Transduction; Signaling Molecule; Surface; System; Systems Biology; Techniques; Technology; Testing; Umbilical Cord Blood; Vaccination; chemokine; cytokine; forward genetics; human disease; macrophage; molecular phenotype; novel; novel strategies; pathogen; programs; response; transcriptomics

Summary The Human Correlation Core will determine whether the novel genes identified by Projects 1 and 2 of this U19 are relevant to human disease. During the preceding fifteen years, the investigators forming the U19 team have identified and contextualized a large number of previously unknown immune regulatory genes, both by forward genetics studies and/or systems biology approaches. In addition, systems approaches have been used to place novel genes within immune regulatory networks. These have included the molecular pathways involved in pathogen recognition, signal transduction, and gene regulation leading to appropriately tailored responses to infection. Over the years, this program has provided seminal information about multiple pathways that mediate inflammation, regulate responses to infection, and underlie successful vaccination. Here the Core will test these genes for relevance in human immunity. The efforts of the Human Correlation Core will be greatly expanded by systems based network analysis (Project 2) and molecular phenotyping (Signaling Core). The Aderem laboratory (Project 2) has established several methods for global transcriptomic analysis of limiting samples including robust amplification protocols for handling small quantities of mRNA and chromatin accessibility measures (ATAC-seq) that can interrogate as few as 500 cells. During the previous funding period of this U19 program, the Signaling Core developed single-cell mass cytometry (CyTOF) as well as methods to both efficiently implement this technology at high- throughput and comprehensively analyze the resulting data. CyTOF permits the simultaneous measurement of 50-100 parameters including surface markers, cytokines and chemokines, and phosphorylated signaling molecules at the single-cell level. Where appropriate, the Core will collaborate with the Aderem and Nolan laboratories to apply these approaches to primary human cells. Since some of our technologies, for example CRISPR/Cas9 gene editing, require large numbers of cells as starting material (it is particularly difficult to use the technique in non-dividing myeloid cells) the Core has established new approaches to isolate large numbers of macrophage lineage cells derived from CD34+ cord blood cells.

概括 人类相关核心将确定该U19项目1和2确定的新基因是否确定 与人类疾病有关。在上一个15年中，调查人员组建了U19团队 已经确定并背景化了大量以前未知的免疫调节基因 正向遗传学研究和/或系统生物学方法。此外，系统方法已经 用于将新型基因放置在免疫调节网络中。这些包括分子途径 参与病原体识别，信号转导和基因调节，导致适当定制 对感染的反应。 多年来，该计划提供了有关中介多种途径的开创性信息 炎症，调节对感染的反应以及成功的疫苗接种。这里的核心将测试 这些基因在人类免疫力方面具有相关性。 基于系统的网络分析，人类相关核心的努力将大大扩展 （项目2）和分子表型（信号芯）。 Aderem实验室（项目2）已建立 用于全局转录组分析的几种限制样本的方法，包括可靠的放大协议 处理少量的mRNA和染色质可及性测量（ATAC-SEQ），可以询问 只有500个细胞。在该U19计划的前几个资金期间，信号核心发展 单细胞质量细胞仪（CytoF）以及有效实施此技术在高处实施该技术的方法 吞吐量并全面分析所得数据。 Cytof允许同时测量 50-100个参数，包括表面标记，细胞因子和趋化因子以及磷酸化信号传导 分子在单细胞水平上。在适当的情况下，核心将与Aderem和Nolan合作 实验室将这些方法应用于原代人细胞。因为我们的一些技术，例如 CRISPR/CAS9基因编辑，需要大量细胞作为起始材料（特别难以使用 非分裂髓样细胞中的技术）核心已经建立了新的方法来隔离大量 衍生自CD34+脐带血细胞的巨噬细胞谱系细胞的。