Microscale System for Functional T Cell Transcriptomics

项目总结/摘要 单细胞分析技术已经改变了对细胞生理学和疾病的理解， 从而深入了解细胞组成和基因组编程的多样性。与此同时， 基于显微镜的测定提供了高水平细胞功能，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 具有特定组学特征的功能将把这两种观点结合成一个引人注目的框架， 了解细胞生理学，但迄今尚未成功实施。为此中央 拟议的研究将结合联合收割机显微镜为基础的分析细胞功能的微图案表面， 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细胞的背景下进行，T细胞是适应性免疫的关键调节剂， 我们的研究小组在从病人身上分离的细胞中观察到细胞迁移的变化， 正在接受慢性淋巴细胞白血病（CLL）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， 转录组学分析，这是目前其他系统不提供的能力。通过这个平台，我们 将进行细胞迁移的变化如何与基因表达的差异相联系的第一次研究， 从而鉴定出与细胞行为相关的特异性转录组标记。根据这些 初步发现，该平台非常适合对复杂的蜂窝系统进行分析 例如从治疗CLL或其它疾病的个体获得的那些。通过提供新的解释 scRNA-Seq数据在细胞功能的背景下，我们设想该系统将导致新的诊断和 改善人类健康的预测工具。

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.