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 单细胞分析技术改变了人们对细胞生理学和疾病的理解， 使人们能够深入了解细胞组成和基因组编程的多样性。同时， 基于显微镜的分析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。将这些复合体联系在一起能力 具有特定术语配置文件的功能将把这两个观点统一到一个引人注目的框架中 了解细胞生理学，但到目前为止还没有成功实施。为此， 拟议的研究将结合基于显微镜的微图案化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了 新的研究途径，并将共同允许将单个细胞的功能与其 转录组。这一新系统将在T细胞的背景下进行，T细胞是适应性免疫的关键调节器， 并受到我们团队从患者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)治疗。这种短期的探索性/发展性 项目将开发第一代平台，能够处理大量的细胞，使当代 转录分析，这是目前其他系统无法提供的功能。有了拟议的平台，我们 将对细胞迁移的差异如何与基因表达的差异进行第一次研究， 从而识别与细胞行为相关的特定转录特征。建立在这些基础上 初步发现，建议平台非常适合对复杂的蜂窝系统进行分析 例如从正在接受慢性淋巴细胞性白血病或其他疾病治疗的个人获得的那些。通过提供新的解释 在细胞功能的背景下，我们预计这个系统将导致新的诊断和 改善人类健康的预测工具。