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 我们提出了单细胞技术来量化提供内部的多参数免疫标记 深度单细胞数据，用于分析我们研究队列中对疫苗接种的免疫反应。这 该服务核心中的共享平台提供了有效且可重复的分析，以支持我们 具有巨大敏感性和细胞类型特异性的研究项目。定义细胞 表型，我们将采用质量细胞术或细胞术（按飞行时间的细胞仪） 多参数单细胞分析，该分析使用重金属离子作为抗体标签并提供 用于免疫亚群细胞分析的巨大细节。进一步概况并关联 具有免疫表型的靶向单细胞的转录组，我们将采用单细胞细胞 通过测序（Cite-seq）进行转录组和表位的索引，以循环血细胞和 通过活检中的确定性条形码（DBIT-seq）的空间分辨率蛋白转录组学 组织（皮肤，淋巴结）。这些转录组学包括BCR和TCR的深度测序 与循环相比，研究组织中的克隆膨胀。识别分子 我们同类群体的疫苗反应的签名，我们将使用质谱（MS）的签名 基于MSTREN的基于印迹的血清蛋白质组学平台，用于无偏见的发现 具有免疫功能的蛋白质是免疫状态的极好反映。 在测定和分析水平上的标准化将允许比较和集成 跨项目的数据。核心C的目标是为 启用从血液和细胞中循环中的免疫细胞的全面表征 可从组织活检中获得，并具有通过cytof和RNA-setering（RNA-Seq）的比较数据， 并匹配蛋白质组学环境的概况。