Core C: Single Cell Phenotyping

核心 C：单细胞表型分析

• 批准号: 10317010
• 负责人: RUTH R MONTGOMERY
• 金额: $ 95.5万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-14 至 2021-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10317010
• 关键词: Antibodies; Behavior; Biological Assay; Biopsy; Blood; COVID-19; 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; 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，该方法将定义明确的队列与公正的大规模结合 分析以阐明在发散疾病的背景下定义免疫反应性的签名 结果。在测定水平和分析水平上的标准化将允许比较和 跨项目的数据集成。为了最大化我们在我们的样品中产生的样品的免疫特征 项目 - 数据的共享分析和解释 - 我们提出了一个单元格表型 使用共享平台进行和分析多参数免疫标记的核心。平台 核心包括用于多参数计的质量细胞仪（Cytof），用于多参数计的单细胞分析。 研究项目（目标1）。这项新技术克服了许多局限 基于荧光的流式细胞术，提供了多个细胞群体前所未有的细胞分析 同样，也是转化研究的强大技术。我们的核心将支持 无缝的Web界面以及项目的深入计算分析需求（AIM 2）。此外， 我们将使用基于纳米威尔的单细胞分析进行多光谱成像细胞仪（音乐）进行测量， 分析和报告在分类种群中的免疫细胞上最多16个标记的表面表达 兴趣或稀疏样品，包括滑液和EM活检（AIM 3）。最后，我们将扩展 纳米韦尔平台执行单细胞RNA序列的功能，提供了独特的 和无偏见的方法来解决免疫小球的异质子集（AIM 4）。整个阵列的RNA- 靶向细胞类型或稀疏样品的测序是探测固有的创新平台 感兴趣的细胞与完善疾病中实施的细胞的分子特征之间的差异。