Multigenerational lineage heterogeneity and metabolic plasticity of CD8 T cells

CD8 T细胞的多代谱系异质性和代谢可塑性

• 批准号: 8681027
• 负责人: SCOTT R MANALIS
• 金额: $ 23.4万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8681027
• 关键词: Autoimmune Diseases; Biochemical Markers; Bypass; CD8B1 gene; Cell surface; Cells; Clinical; Environment; Fluorescent Antibody Technique; Generations; Growth; Heterogeneity; Immune response; Individual; Label; Liquid substance; Maps; Measurement; Measures; Memory; Metabolic; Metabolism; Metric; Microfluidics; Monitor; Pharmaceutical Preparations; Phenotype; Play; Population; Process; Relative (related person); Role; Shapes; Sirolimus; Solutions; Staging; Staining method; Stains; Surface; T cell differentiation; T-Cell Activation; T-Cell Development; T-Lymphocyte; Time; Vaccination; cancer immunotherapy; cell growth; insight; meetings; novel; public health relevance; response; sensor; vaccination strategy

DESCRIPTION (provided by applicant): From a clinical perspective, understanding and manipulating the dynamics of T cell development may offer key insight in to the progression of various autoimmune diseases as well as present potential treatment options with regards to cancer immunotherapy and vaccination. Although existing single cell approaches are now elucidating the significance of heterogeneity, the way in which T cells evolve to achieve particular phenotypic states remains unclear. We are proposing to use novel microfluidic platforms for dynamically monitoring the response of individual CD8+ T cells over several generations to investigate differentiation, plasticity and metabolism. To accomplish this, we will utilize hydrodynamic traps to dynamically interrogate several generations of a single T cell's progeny by implementing standard immunofluorescence techniques on-chip. Since fluid surrounding the cells can be rapidly and frequently exchanged without perturbing growth, we will be able to monitor expression of cell surface markers and deliver drugs that alter cellular metabolism at precise time points. As a compliment to biochemical markers of lineage we will also measure multigenerational growth rates of single activated T cells in order to provide a physical measurement of the cell's metabolic state.

描述（由申请人提供）：从临床角度来看，理解和操纵 T 细胞发育的动态可能为了解各种自身免疫性疾病的进展提供关键见解，并提供有关癌症免疫治疗和疫苗接种的潜在治疗选择。尽管现有的单细胞方法现在正在阐明异质性的重要性，但 T 细胞进化以实现特定表型状态的方式仍不清楚。我们建议使用新型微流体平台动态监测单个 CD8+ T 细胞几代的反应，以研究分化、可塑性和代谢。为了实现这一目标，我们将通过在芯片上实施标准免疫荧光技术，利用流体动力学陷阱动态询问单个 T 细胞后代的几代。由于细胞周围的液体可以快速、频繁地交换，而不会干扰生长，因此我们将能够监测细胞表面标志物的表达，并在精确的时间点提供改变细胞代谢的药物。作为对谱系生化标记的补充，我们还将测量单个活化 T 细胞的多代生长速率，以便提供细胞代谢状态的物理测量。