Multi-omic immune profiling

多组学免疫分析

• 批准号: 10617755
• 负责人: RUTH R MONTGOMERY
• 金额: $ 53.15万
• 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 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10617755
• 关键词: Adaptive Immune System; Antibodies; B-Cell Antigen Receptor; B-cell receptor repertoire sequencing; Bar Codes; Biological Assay; Biopsy; Blood; Blood Cells; Blood specimen; Cell Surface Proteins; Cells; Cellular Indexing of Transcriptomes and Epitopes by Sequencing; Characteristics; Circulation; Clinical; Clonal Expansion; Cohort Studies; Collaborations; Complex; Cytometry; Data; Data Analyses; Detection; Dimensions; Disease; Frequencies; Gene Expression; Gene Expression Profile; Genes; Genetic Transcription; Goals; Heavy Metals; Immune; Immune response; Immunologic Markers; Immunologics; Immunophenotyping; Innate Immune System; Investigation; Ions; Knowledge; Label; Manuals; Maps; Mass Spectrum Analysis; Measurement; Membrane Proteins; Messenger RNA; Molecular; Molecular Profiling; National Institute of Allergy and Infectious Disease; Pathogenesis; Pathway interactions; Peripheral Blood Mononuclear Cell; Phenotype; Plasma; Population; Process; Proteins; Proteome; Proteomics; Quality Control; Reproducibility; Research Project Grants; Resolution; Resources; Sampling; Serum; Services; Signal Pathway; Signal Transduction; Site; Skin; Specificity; Standardization; T-Lymphocyte; Technology; Time; Tissue Sample; Tissues; Vaccination; Vaccines; Visualization; Vulnerable Populations; Weight; biomarker discovery; cell type; clinically relevant; cohort; comparative; computerized tools; data integration; data management; data repository; dimensional analysis; feeding; functional status; high dimensionality; human disease; immune function; immunological status; insight; interest; lymph nodes; multiple omics; new technology; protein expression; response; response biomarker; sharing platform; single cell analysis; single cell technology; study population; transcriptome; transcriptome sequencing; transcriptomic profiling; transcriptomics; vaccine response

Abstract We propose single-cell technologies to quantify multiparameter immune markers that provide in- depth single-cell data for analysis of immune responses to vaccination in our study cohorts. The shared platforms in this service core provide efficient and reproducible profiling in support of our research projects with greatly expanded sensitivity and cell-type specificity. To define cell phenotypes, we will employ mass cytometry or CyTOF (Cytometry by Time-Of-Flight) for multiparameter single-cell analysis, which uses heavy metal ions as antibody labels and provides tremendous detail for cellular analysis of immune subsets. To further profile and correlate the transcriptomes of targeted single cells with immunophenotype, we will employ single-cell cellular indexing of transcriptomes and epitopes by sequencing (CITE-Seq) for circulating blood cells and spatial-resolution proteo-transcriptomics via deterministic barcoding (DBiT-seq) in biopsies from tissues (skin, lymph node). These transcriptomics include in depth sequencing of BCR and TCR to investigate clonal expansion in the tissue compared to the circulation. To identify molecular signatures of vaccine responses across our cohorts, we will use a mass spectrometry (MS)-based MStern blotting-based serum proteomics platform for unbiased discovery of several hundred proteins with immunological function as an excellent reflection of the immunological state. Standardization, both at the assay and analytical levels, will allow comparisons and integration of data across projects. The goals of Core C are to provide a unique and essential resource to enable the comprehensive characterization of circulating immune cells from blood as well as cells available from tissue biopsies, with comparative data by CyTOF and RNA-sequencing (RNA-seq), and matched profiles of proteomic milieu.

抽象的 我们提出单细胞技术来量化多参数免疫标记物，这些标记物提供了以下信息： 深度单细胞数据，用于分析我们研究队列中疫苗接种的免疫反应。这 该服务核心中的共享平台提供高效且可重复的分析，以支持我们的 大大扩展了敏感性和细胞类型特异性的研究项目。定义单元格 表型，我们将采用质谱流式细胞术或 CyTOF（飞行时间细胞术）进行分析 多参数单细胞分析，使用重金属离子作为抗体标记，并提供 免疫亚群细胞分析的大量细节。为了进一步分析和关联 具有免疫表型的目标单细胞的转录组，我们将采用单细胞细胞 通过循环血细胞测序 (CITE-Seq) 对转录组和表位进行索引 通过确定性条形码 (DBiT-seq) 在活检中进行空间分辨率蛋白质转录组学 组织（皮肤、淋巴结）。这些转录组学包括 BCR 和 TCR 的深度测序 研究组织中与循环相比的克隆扩增。鉴定分子 为了确定我们队列中疫苗反应的特征，我们将使用基于质谱 (MS) 的方法 基于 MStern 印迹的血清蛋白质组学平台，可公正地发现数百种蛋白质 具有免疫功能的蛋白质可以很好地反映免疫状态。 化验和分析水平上的标准化将允许比较和整合 跨项目的数据。 Core C 的目标是提供独特且重要的资源 能够全面表征血液和细胞中的循环免疫细胞 可从组织活检中获取，并通过 CyTOF 和 RNA 测序 (RNA-seq) 获得比较数据， 以及蛋白质组环境的匹配概况。