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: 8896418
• 负责人: SCOTT R MANALIS
• 金额: $ 19.5万
• 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 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8896418
• 关键词: Autoimmune Diseases; Biochemical Markers; Bypass; CD8B1 gene; Cell surface; Cells; Clinical; Environment; Fluorescent Antibody Technique; Generations; Growth; Health; Heterogeneity; Immune response; Individual; Label; Liquid substance; Maps; Measurement; Measures; Memory; Metabolic; Metabolism; 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; adaptive immunity; cancer immunotherapy; cell growth; insight; meetings; novel; 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会为各种自身免疫性疾病的发展提供关键的见解，以及有关癌症免疫疗法和疫苗接种的潜在治疗选择。尽管现有的单细胞方法现在阐明了异质性的重要性，但T细胞发展以实现特定表型态的方式尚不清楚。我们建议使用新型的微流体平台，以动态监测几代人的单个CD8+ T细胞的响应，以研究分化，可塑性和代谢。为此，我们将利用流体动力陷阱来通过片上实现标准的免疫荧光技术来动态询问几代单个T细胞的后代。由于周围细胞周围的液体可以迅速且经常交换而不会扰动生长，因此我们将能够监测细胞表面标记的表达并输送在精确时间点改变细胞代谢的药物。为了称赞谱系的生化标志物，我们还将测量单个激活T细胞的多代增长率，以便对细胞的代谢状态进行物理测量。