Microscale System for Functional T Cell Transcriptomics

功能性 T 细胞转录组学微型系统

• 批准号: 10156792
• 负责人: Lance C Kam
• 金额: $ 23.34万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-16 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10156792
• 关键词: Address; Basic Science; Biological Assay; Biological Models; CD28 gene; CD3 Antigens; CD4 Positive T Lymphocytes; Cell physiology; Cells; Chemicals; Chronic Lymphocytic Leukemia; Complex; Data; Development; Disease; Disease Progression; Environment; Gene Expression; Gene Expression Profile; Generations; Genomics; Health; Human; Immune response; In Vitro; Individual; Individuality; Investigation; Knowledge; Ligands; Link; Malignant Neoplasms; Measures; Microfabrication; Microfluidics; Microscopy; Minor; Mus; Natural regeneration; Patients; Performance; Physiological; Physiology; Population; Positioning Attribute; Production; Proteomics; Source; Stimulus; Surface; System; T-Lymphocyte; Techniques; Technology; Testing; Tissues; Variant; Vision; adaptive immunity; base; cell behavior; cell motility; design; exhaustion; extracellular; fundamental research; human disease; improved; indexing; individual response; individual variation; individualized medicine; insight; lithography; migration; novel diagnostics; prognostic tool; programmed cell death protein 1; prototype; receptor; response; single cell analysis; single-cell RNA sequencing; therapy development; transcriptome; transcriptomics

Project Summary / Abstract Single cell analysis technologies have transformed understanding of cellular physiology and disease, allowing deep insights into the diversity of cell composition and genomic programming. Concurrently, microscopy-based assays provide individualized measures of high-level cellular function, such as migration, secretion of chemical factors, and interaction with neighboring cells. The ability to associate these complex functions with specific -omic profiles would unite these two perspectives into a compelling framework for understanding cellular physiology, but to date has not been successfully implemented. To this end, the proposed study will combine microscopy-based analysis of cell function on micropatterned surfaces with an emerging platform for single cell RNA sequencing (scRNA-Seq). These two platforms have individually led to new avenues of investigation, and together will allow association of an individual cell’s function with its transcriptome. This new system will be pursued in the context of T cells, key modulators of adaptive immunity, and is inspired by the variation in cell migration our team has observed in cells isolated from patients undergoing treatment for chronic lymphocytic leukemia (CLL). This short-term, exploratory / developmental project will develop a first generation platform capable of processing numbers of cells that allow contemporary transcriptomic analysis, a capability not currently provided by other systems. With the proposed platform, we will carry out the first studies of how variation in cell migration is linked to differences in gene expression, leading to identification of specific transcriptomic signatures associated with cell behavior. Building on these initial discoveries, the proposal platform is well positioned to carry out analysis on complex cellular systems such as those obtained from individuals being treated for CLL or other disease. By providing new interpretation of scRNA-Seq data in the context of cell function, we envision that this system will lead to new diagnostic and prognostic tools to improve human health.

项目摘要 /摘要 单细胞分析技术已经改变了对细胞生理和疾病的理解， 可以深入了解细胞组成和基因组编程的多样性。同时 基于显微镜的测定提供了对高级细胞功能（例如迁移）的个性化测量值 化学因子的分泌以及与邻近细胞的相互作用。将这些复合物关联的能力 具有特定 - 元素概况的功能将把这两个观点团结成一个引人入胜的框架 了解细胞生理学，但迄今为止尚未成功实施。为此， 拟议的研究将结合基于显微镜在微图表上的细胞功能的分析 单细胞RNA测序（SCRNA-SEQ）的新兴平台。这两个平台已单独导致 新的投资途径，并将共同使单个单元的功能与其联系 转录组。该新系统将在T细胞（适应性免疫组织化学的关键调节剂）的背景下追求 并受到细胞迁移的差异的启发，我们的团队在从患者中分离的细胞中观察到 接受慢性淋巴细胞性白血病（CLL）的治疗。这种短期的探索 /发展 项目将开发一个能够处理允许当代细胞数量的第一代平台 转录组分析，其他系统目前未提供的功能。使用提出的平台，我们 将对细胞迁移的变异如何与基因表达差异有关的第一批研究， 导致鉴定与细胞行为相关的特定转录组特征。建立在这些基础上 最初的发现，该提案平台可以很好地对复杂的蜂窝系统进行分析 例如从接受CLL或其他疾病治疗的个体获得的那些。通过提供新的解释 在单元函数的背景下，SCRNA-Seq数据的数据，我们设想该系统将导致新的诊断和 改善人类健康的预后工具。