Microscale System for Functional T Cell Transcriptomics

功能性 T 细胞转录组学微型系统

• 批准号: 10319619
• 负责人: Lance C Kam
• 金额: $ 19.67万
• 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 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10319619
• 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; diagnostic tool; 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.

项目概要/摘要 单细胞分析技术改变了对细胞生理学和疾病的理解， 允许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高水平细胞功能的个性化测量，例如迁移、 化学因子的分泌以及与邻近细胞的相互作用。将这些复杂的事物联系起来的能力 具有特定-omic配置文件的功能将把这两种观点结合成一个令人信服的框架 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的微图案表面细胞功能分析与 新兴的单细胞 RNA 测序平台 (scRNA-Seq)。这两个平台分别导致 新的研究途径，并且共同将允许将单个细胞的功能与其 转录组。这个新系统将在 T 细胞（适应性免疫的关键调节剂）的背景下进行研究。 并受到我们团队在从患者身上分离的细胞中观察到的细胞迁移变化的启发 正在接受慢性淋巴细胞白血病（CLL）的治疗。这种短期的、探索性的/发展性的 该项目将开发第一代平台，能够处理大量细胞，使当代 转录组分析，这是其他系统目前不提供的功能。通过所提议的平台，我们 将首次研究细胞迁移的变化如何与基因表达的差异相关， 导致鉴定与细胞行为相关的特定转录组特征。在此基础上 初步发现，该提案平台非常适合对复杂的细胞系统进行分析 例如从正在接受 CLL 或其他疾病治疗的个体获得的数据。通过提供新的解释 细胞功能背景下的 scRNA-Seq 数据，我们预计该系统将带来新的诊断和 改善人类健康的预测工具。