Core C: Single Cell Phenotyping

核心 C：单细胞表型分析

• 批准号: 10079820
• 负责人: RUTH R MONTGOMERY
• 金额: $ 44.97万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-12 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10079820
• 关键词: Antibodies; Behavior; Biological Assay; Biopsy; Blood; Cells; Cerebrospinal Fluid; Clinical; Collaborations; Computer Analysis; Custom; Cytometry; Data; Data Analyses; Data Management Resources; Detection; Disease; Disease Outcome; Flow Cytometry; Fluorescence; Gene Expression Profile; Genetic Transcription; Goals; Heavy Metals; Human; Image Cytometry; Immune; Immune response; Immune system; Immunologic Markers; Immunologics; Immunophenotyping; Infection; Investigation; Ions; Label; Measurement; Measures; Molecular Profiling; National Institute of Allergy and Infectious Disease; Pathogenesis; Pathway interactions; Patients; Phenotype; Population; Reporting; Reproducibility; Research Project Grants; Resources; Sampling; Services; Signal Pathway; Signal Transduction; Specificity; Sputum; Standardization; Surface; Synovial Fluid; System; Techniques; Technology; Time; Tissues; Visualization; base; cell type; cohort; comparative; computerized tools; data integration; data management; data repository; dimensional analysis; erythema migrans; exhaustion; feeding; high dimensionality; human disease; innovation; interest; new technology; pathogen; response; sharing platform; single cell analysis; single cell technology; single-cell RNA sequencing; targeted sequencing; transcriptome; transcriptome sequencing; web interface

Abstract We propose to use a systems approach that combines well-defined cohorts with unbiased large-scale profiling to elucidate signatures defining immune responsiveness in the context of divergent disease outcomes. Standardization, both at the assay and analytical levels, will allow comparisons and integration of data across projects. To maximize the immune profiles of samples generated in our Projects—and the shared analysis and interpretation of data— we propose a Single Cell Phenotyping Core to conduct and analyze multiparameter immune markers using shared platforms. The platforms of the Core include mass cytometry (CyTOF) for multiparameter single cell analysis of samples from the Research Projects (Aim 1). This new technology, which overcomes many of the limitations of fluorescence-based flow cytometry, provides unprecedented cellular analysis of multiple cell populations simultaneously and is a powerful technique for translational investigations. Our core will support seamless web interface and in depth computational analysis needs of the projects (Aim 2). In addition, we will use nanowell-based single-cell analysis for multispectral imaging cytometry (MuSIC) to measure, analyze and report the surface expression of up to 16 markers on immunocytes in sorted populations of interest or in sparse samples, including synovial fluid and EM biopsies (Aim 3). Finally, we will extend the capabilities of the nanowell platform for performing single-cell RNA-sequencing, which provides a unique and unbiased approach for resolving heterogeneous subsets of immune cells (Aim 4). Array-wide RNA- sequencing of targeted cell types or sparse samples is an innovative platform to probe the intrinsic differences between cells of interest and to refine the molecular signatures of cells implicated in disease.

抽象的 我们建议使用一种系统方法，将明确定义的群体与公正的大规模研究相结合 分析以阐明不同疾病背景下定义免疫反应的特征 结果。化验和分析水平上的标准化将允许进行比较和 跨项目的数据集成。为了最大化我们生成的样本的免疫特征 项目——以及数据的共享分析和解释——我们提出了单细胞表型分析 使用共享平台进行和分析多参数免疫标记物的核心。的平台 核心包括质谱流式分析 (CyTOF)，用于对来自以下来源的样品进行多参数单细胞分析 研究项目（目标 1）。这项新技术克服了许多限制 基于荧光的流式细胞术，为多个细胞群提供前所未有的细胞分析 同时也是一种强大的转化研究技术。我们的核心将支持 无缝网络界面和项目的深入计算分析需求（目标 2）。此外， 我们将使用基于纳米孔的单细胞分析进行多光谱成像细胞术（MuSIC）来测量， 分析并报告分类群体中免疫细胞上多达 16 个标记的表面表达 对稀疏样本感兴趣或感兴趣，包括滑液和电镜活检（目标 3）。最后，我们将延长 纳米井平台执行单细胞 RNA 测序的能力，提供了独特的 以及解决免疫细胞异质子集的公正方法（目标 4）。全阵列 RNA- 目标细胞类型或稀疏样本的测序是探索内在特征的创新平台 感兴趣的细胞之间的差异并细化与疾病有关的细胞的分子特征。